Methods in Enzymology Vol 199: Cumulative Subject Index, Volumes 168-174, 176-194

Preface The idea for a cumulative index was recognized by the founding editors who prepared one for Volumes I through VI...

Author: John N. Abelson | Melvin I. Simon

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Preface The idea for a cumulative index was recognized by the founding editors who prepared one for Volumes I through VI of Methods in Enzymology by weeding and interpolating from the entries that had been indexed in the individual volumes. As the series developed in both number and complexity, different individuals with different backgrounds served as volume indexers. Subsequently, the series was fortunate in having Dr. Martha G. Dennis and Dr. Edward A. Dennis accept the challenge of computerizing the data available from these individual indexes, and this effort resulted in Volumes 33, 75, and 95, which cover Volumes 1 through 80. Although each of the three books produced with the aid of computerization provided an appropriate cumulative index, major problems were encountered. One was time, both expensive computer time and lag time before such efforts resulted in publication. The most important difficulty was that the compilers were hampered by the lack of uniformity in the indexing of the individual volumes, resulting in the need for much hand editing to achieve a reasonable collation. The products were very decent. if uneven, indexes that also contributed to the methodology of computerized indexing, albeit with much delay and great expense. This cumulative index has been produced by the staff of Academic Press. The indexers have gone back to the text rather than to the individual volume indexes and, using a uniform set of guidelines, have culled the major topics, leading to five to ten entries for each article. Thus, if searching by name for one of the dozen substrates of an enzyme, the isolation of which is being presented, it probably will not be found in the index, although the assay substrate may be there. Nor will the specific inhibitors of the enzyme be itemized, although the topic of the enzyme's inhibition will form an entry. The index, then, is not complete, but should lead to the broad subject headings. Since there is a tendency to identify specific topics and methods with particular individuals, a contributor index is included. Finally, the complete table of contents of each of the volumes indexes is included.

vii

Contents of Volumes 168-174, 176-194

VOLUME 168 HORMONE ACTION (PART K: NEUROENDOCRINE PEPTIDES)

S e c t i o n I. P r e p a r a t i o n

of Chemical Probes

1. Strategy and Methodology for Development of Antisera against Procholecystokinin MARGERY C. BEINFELD AND THOMAS L. O'DONOHUE 2. High-Density Multiple Antigen-Peptide System for Preparation of Antipeptide Antibodies JAMES P. TAM 3. Preparation and Use of Complementary Peptides KENNETH L. BOST AND J. EDWIN BLALOCK 4. Fluorescent and Cytotoxic Analogs of Corticotropin-Releasing Factor: Probes for Studying Target Cells in Heterogeneous Populations JEFFREY SCHWARTZ AND WYLIE VALE

S e c t i o n II. E q u i p m e n t

7 16

29

and Technology

5. Cell-to-Cell Communication in Peptide Target Cells of Anterior Pituitary CARL DENEF, PHILIPPE MAERTENS, WILFRIED ALLAERTS, ANNICK MIGNON, WIM ROBBERECHT, LUC SWENNEN, AND PETER CARMELIET 6. Application of Fast Atom Bombardment Mass Spectrometry to Posttranslational Modifications of Neuropeptides P . C . ANDREWS AND JACK E. DIXON 7. Electrophysiological Analysis of Opioid Peptides: Extracellular and lntracellular Approaches EDWARD D. FRENCH AND JOHN T. WILLIAMS 8. Dose-Response Testing of Peptides by Hippocampal Brain Slice Recording M. IAN PHILLIPS AND REINHARD A. PALOVEIK 9. Patch Clamp Recording from Anterior Pituitary Cells Identified by Reverse Hemolytic Plaque Assay THOMAS L. CROXTON, WILLIAM McD. ARMSTRONG, AND NIRA BEN-JONATHAN 10. Microdialysis for Measurement of Neuroendocrine Peptides JON E. LEVINE AND KEITH D. POWELL I I. Use of Microdialysis in Neuroendocrinology KEITH M. KENDRICK 12. In Vitro Perifusion of Human Hypothalamic and Pituitary Tissue DENNIS D. RASMUSSEN 13. Transient Permeabilization of Endocrine Cells: lnositol Lipid Metabolism IRENE WINICOV AND MARVIN C. GERSHENGORN 14. Cell Cracking: Permeabilizing Cells to Macromolecular Probes THOMAS F. J. MARTIN ix

3

47 72 103 129

144 166 182 206 218 225

CONTENTS OF VOLUMES 168-174, 176-194

X

15. Microsurgical Techniques for Studying Functional Correlates of Hypothalamohypophyseal Axis ICHIRO MURAI, WALTER C. LOW, AND NIRA BEN-JONATHAN

234

16. Superfusion and Static Culture Techniques for Measurement of Rapid Changes in Prolactin Secretion GONZALO MART~NEZ DE LA ESCALERA, KAREN C. SWEARINGEN, AND RICHARD I. WEINER

254

17. Intracellular Calcium Levels in Rat Anterior Pituitary Cells: Single-Cell Techniques DENIS A. LEONG

263

S e c t i o n III. P r e p a r a t i o n a n d M a i n t e n a n c e o f Biological M a t e r i a l s 18. Use of Protein Kinase C-Depleted Cells for Investigation of the Role of Protein Kinase C in Stimulus-Response Coupling in the Pituitary CRAIG A. MCARDLE AND P. MICHAEL CONN 19. Purification of Neuropeptides: Cholecystokinin and Vasoactive Intestinal Peptide JOHN ENG AND ROSALYNS. YALOW 20. Purification and Characterization of Insulin-Like Growth Factor-II Receptors CAROLYN D. SCOTT AND ROBERT C. BAXTER

287

302 309

S e c t i o n IV. Q u a n t i t a t i o n of N e u r o e n d o c r i n e S u b s t a n c e s 21. Measurement of Hormone Secretion from Individual Cells by Cell Blot Assay MARCIA E. KENDALLAND W. C. HYMER 22. Measurement of Phosphoinositide Hydrolysis in Isolated Cell Membrane Preparations MILLIE M. CHIEN AND JOHN C. CAMBIER 23. A Rapid Method for the Resolution of Protein Kinase C Subspecies from Rat Brain Tissue

MARK S. SHEARMAN, KOUJI OGITA, USHIO KIKKAWA, AND YASUTOMI NISHIZUKA

327 338

347

24. Assay of Peptidylglycine Monooxygenase: Glycine-Directed Amidating Enzyme THOMAS BROCK, JANE HUMM, AND J. S. KIZER

25. Assay of Glutaminylpeptide Cyclase JEANNE B. KOGER, JANE HUMM, AND JOHN S. KIZER 26. Primary Thyrotropin-Releasing Hormone-Degrading Enzymes CHARLES H. EMERSON 27. Quantification of the Mass of Tyrosine Monooxygenase in the Median Eminence and Superior Cervical Ganglion JOHN C. PORTER, PAULUS S. WANG, WOJCIECH KEDZIERSKI, AND HECTOR A. GONZALEZ

28. Measurement of Vasopressin-Converting Aminopeptidase Activity and Vasopressin Metabolites J . P . H . BURaACHAND BIN LIU 29. Quantitation of Vasopressin and Oxytocin mRNA Levels in the Brain HUBERT H. M. VAN TOL AND J. PETER H. BURBACH 30. Granulosa Cell Aromatase Bioassay for Follicle-Stimulating Hormone KRISTINE D. DAHL, XIAO-CHI JIA, AND AARON J. W. HSUEH 31. Neuroendocrine Regulation of Oocyte Tissue Plasminogen Activator THOM.ASA. BICSAK,CARLAM. HEKMAN, AND AARONJ. W. HSUEH

351 358

365

371

385 398 414

422

xi

CONTENTS OF VOLUMES 168-174, 1 7 6 - 1 9 4

32. Assessment of Peptide Regulation of the Autonomic Nervous System MARVIN R. BROWN, ROBERTA ALLEN, AND LAUREL A. FISHER

431

33. Recognition, Purification, and Structural Elucidation of Mammalian Physalaemin-Related Molecules LAWRENCE H. LAZARUS AND WILLIAM E. WILSON

444

34. Radioligand-Binding Assays for Study of Neurotensin Receptors MICHEL GOEDERT

462

35. Characterization of Receptors for Bombesin/Gastrin-Releasing Peptide in Human and Murine Cells TERRY W. MOODY, RICHARD M. KRIS, GARY FISKUM, CAROL D. LINDEN, M. BERG, AND JOSEPH SCHEESSINGER

481

36. Isolation and Identification of Neuroendocrine Peptides from Milk ELI HAZUM

494

37. Passive Immunoneutralization: A Method for Studying the Regulation of Basal and Pulsatile Hormone Secretion MICHAEL D. CULLER AND ANDRI~S NEGRO-VILAR

498

38. Combined Antibody-High-Performance Liquid Chromatography Approach to Assess Prohormone Processing W. WETSEL AND A. NEGRO-VILAR 39. Chromatographic Methods for Characterization of Angiotensin in Brain Tissue

517

KLAUS HERMANN, MOHAN K. RAIZADA, AND M. IAN PHILLIPS

545

40. Measurement of Thyrotropin-Releasing Hormone and Its Metabolites CHARLES H. EMERSON

567

GuY VALIQUETTE AND SIMON NEUBORT

574

41. Monoclonal Antibodies: Uses in Studies on Vasopressin 42. Detection and Purification of Inhibin Using Antisera Generated against Synthetic Peptide Fragments JOAN M. VAUGHAN, JEAN RIVIER, ANNE Z. CORRIGAN, RICHARD McCLINTOCK, CAROLYN A. CAMPEN, DIANE JOLLEY, JOSEF K. VOGLMAYR, C. WAYNE BARDIN, CATHERINE RIVIER, AND WYLIE VALE

43. Regulation of Neuroendocrine Peptide Gene Expression M. BLUM 44. Solution Hybridization-Nuclease Protection Assays for Sensitive Detection of Differentially Spliced Substance P- and Neurokinin A-Encoding Messenger Ribonucleic Acids JAMES E. KRAUSE, JEAN D. CREMINS, MARK S. CARTER, ELAINE R. BROWN, AND MARGARET R. MACDONALD

45. Quantifying Carrier-Mediated Transport of Peptides from the Brain to the Blood WILLIAM A. BANKS AND ABBA J. KASTIN 46. Characterizing Molecular Heterogeneity of Gastrin-Releasing Peptide and Related Peptides JOSEPH R. REEVE, JR. AND JOHN H. WALSH

588

618

634

652 660

S e c t i o n V. U s e o f C h e m i c a l P r o b e s 47. Neuropeptide Gene Transcription in Central and Peripheral Nervous System Tissue by Nuclear Run-On Assay JEFFREY D. WHITE AND EDMUND F. LAGAMMA 48. Assaying the Reporter Gene Chloramphenicol Acetyltransferase

681

DAVID W. CRABB, CAROLYN D. MINTH, AND JACK E. DIXON

690

49. In Situ Hybridization Histochemical Detection of Neuropeptide mRNA Using DNA and RNA Probes W. SCOTT YOUNG III 50. Use of Metal Complexes in Neuroendocrine Studies AYALLA BARNEA

702 710

xii

CONTENTS OF VOLUMES 168-174, 176-194

51. Methods for the Study of Somatostatin M.C. AGUILAAND S. M. McCANN 52. Mapping of Gonadotropin-Releasing Hormone-Receptor Binding Site Using Selective Chemical Modifications ELI HAZUM

716 729

S e c t i o n VI. L o c a l i z a t i o n of N e u r o e n d o c r i n e S u b s t a n c e s 53. In Situ Hybridization: Quantitation Using Radiolabeled Hybridization Probes GEORGE R. UHL 54. Nonradioactive Methods of in Situ Hybridization: Visualization of Neuroendocrine mRNA P. C. EMSON, H. ARAI, S. AGRAWAL, C. CHRISTODOULOU, AND M. J. GAIT 55. High-Resolution in Situ Hybridization Histochemistry FRANK BALDINO, JR., MARIE-FRANCOISE CHESSELET, AND MICHAEL E. LEWIS

56. In Situ Hybridization Histochemistry Combined with Markers of Neuronal Connectivity JAMES S. SCHWABER, BIBIE M. CHRONWALL, AND MICHAEL E. LEWIS 57. Cytochemical Techniques for Studying the Diffuse Neuroendocrine System A. E. BISHOPAND J. M. POLAK 58. Quantitation and Digital Representation of in Situ Hybridization Histochemistry MICHAEL E. LEWIS, WADE T. ROGERS, RUDOLPH G. KRAUSE II, AND JAMES S. SCHWABER

741

753

761

778

791

808

59. Graphical and Statistical Approaches to Data Analysis for in Situ Hybridization JOSEPH T. MCCABE, ROBERT A. DESHARNAIS, AND DONALD W. PFAFF

822

V O L U M E 169 PLATELETS: RECEPTORS, ADHESION, SECRETION

S e c t i o n I. Isolation o f P l a t e l e t s 1. Isolation of Human Platelets from Plasma by Centrifugation and Washing J. FRASER MUSTARD, RAELENE L. KINLOUGH-RATHBONE,AND MARIAN A. PACKHAM

3

2. Isolation of Human Platelets by Albumin Gradient and Gel Filtration SHEILA TIMMONS AND JACEK HAWIGER

l1

3. Isolation and Characterization of Porcine Platelets BOGUSLAW RUCINSKI AND STEFAN NIEWlAROWSKI

22

4. Isolation of Platelets from Laboratory Animals JAMES L. CATALFAMOAND W. JEAN DODDS

27

S e c t i o n II. P l a t e l e t A d h e s i o n , A g g r e g a t i o n , an d T u r n o v e r 5. Measurements of Platelet Interaction with Components of the Vessel Wall in Flowing Blood KJELL S. SAKARIASSEN,RETO MUGGLI, AND HANS R. BAUMGARTNER

37

CONTENTS OF VOLUMES 168--174, 176-194 6. Measurement of Platelet Interaction with Endothelial Monolayers GLENNA L. FRY AND JOHN C. HOAK 7. Measurement of Platelet and Megakaryocyte Interaction with the Subendothelial Extracellular Matrix AMIRAM ELDOR, ZVI FUKS, RICHARD F. LEVlNE, AND ISRAEL VLODAVSKY 8. EX Vivo Measurement of Platelet Adhesion to Polymeric Surfaces K1NAM PARK, DEANE F. MOSHER, AND STUART L. COOPER 9. Platelet Interaction with Artificial Surfaces: In Vitro Evaluation JACK N. LINDON, LESLIE KUSHNER, AND EDWIN W. SALZMAN 10. Platelet Aggregation Measured by the Photometric Method MARJORIE B. ZUCKER 11. Platelet Aggregation Measured in Vitro by Microscopic and Electronic Particle Counting MONYM. FROJMOVIC, JOHN G. MILTON, AND ADRIAN L. GEAR 12. Fixation of Platelets and Platelet Agglutination/Aggregation Tests K. M. BRINKHOUS AND MARJORIE S. READ 13. Determination of Platelet Volume and Number DAVID BESSMAN 14. Measurement of in Vivo Platelet Turnover and Organ Distribution Using tEqnLabeled Platelets W. ANDREW HEATON, ANTHON DU P. HEYNS, AND J. HEINRICH JOIST

xiii 71

76 91 104 117 134 149 164

172

S e c t i o n III. P l a t e l e t S e c r e t i o n 15. Platelet Secretory Pathways: An Overview

JACEK HAWIGER

191

A. Dense Granule Constituents 16. Measurement of Secretion of Adenine Nucleotides HOLM HOLMSEN AND CAROL A. DANGELMAIER 17. Measurement of Secretion of Serotonin HOLM HOLMSEN AND CAROL A. DANGELMAIER

195 205

B. a-Granule Constituents 18. Platelet-Derived Growth Factor: Purification and Characterization HARRY N. ANTONIADES AND PANAYOTIS PANTAZIS 19. Platelet Basic Protein, Low-Affinity Platelet Factor 4, and/~-Thromboglobulin: Purification and Identification JOHN C. HOLT AND STEFAN NIEWIAROWSKI 20. Platelet a-Granule Protein-Induced Chemotaxis of Inflammatory Cells and Fibroblasts ROBERT M. SENIOR, GAIE L. GRIFFIN, AKIRO KIMURA, JUNG SAN HUANG, AND THOMAS F. DEUEL 21. Secretion of von Willebrand Factor from Platelets MARIA FERNANDA LOPEZ-FERNANDEZ, JAVIER BATLLE, ZAVERIO M. RUGGERI, AND THEODORE S. ZIMMERMAN 22. Secretion of Thrombospondin from Human Blood Platelets HENRY S. SLAYTER 23. Platelet Histidine-Rich Glycoprotein LAWRENCE L. K. LEUNG, PETER C. HARPEL, AND RALPH L. NACHMAN

210

224

233

244 251 268

xiv

CONTENTS OF VOLUMES 168-174, 176-194

24. Platelet High-Molecular-Weight Kininogen ALVIN H. SCHMAIER AND ROBERT W. COLMAN 25. Platelet a2-Antiplasmin EDWARD F. PLOW, LINDSEY A. MILES, AND DESIRE COLLEN 26. Immunocytochemical Localization of Platelet Granule Proteins JAN J. SIXMA, JAN-WILLEM SLOT, AND HANS J. GEUZE 27. PADGEM Protein CINDY L. BERMAN, ERIK L. YEO, BARBARa C. FURIE, AND BRUCE FURIE 28. Dark-Field Electron Microscopy of Platelet Adhesive Macromolecules HENRY S. SLAYTER

276 296 301 311 326

C. Lysosomal Enzymes 29. Measurement of Secretion of Lysosomal Acid Glycosidases HOLM HOLMSEN AND CAROL A. DANGELMAIER 30. Platelet Heparitinase ROBERT D. ROSENaERG

336 342

Section IV. Regulation of Platelet Function 31. Interaction of Extracellular Calcium with the Surface Membrane of Human Platelets LAWRENCE F. BRASS AND SANFORD J. SHATTIL 32. Calcium-45 Exchange Techniques to Study Calcium Transport in Intact Platelets LAWRENCE F. BRASS AND ELIZABETH BELMONTE 33. Measurement of Platelet Cytoplasmic Ionized Calcium Concentration with Aequorin and Fluorescent Indicators PETER C. JOHNSON, J. ANTHONY WARE, AND EDWIN W. SALZMAN 34. Endogenous Phosphatidylinositol 4,5-Bisphosphate, Phospbatidylinositol, and Phosphatidic Acid in Stimulated Human Platelets M. JOHAN BROEKMAN 35. Membrane-Permeable Diacylglycerol, Its Application to Platelet Secretion, and Regulation of Platelet Protein Kinase C NORIO KAJIKAWA, USHIO KIKKAWA, KIYOSH! ITOH, AND YASUTOMI NlSHIZUKA 36. Calcium-Dependent Proteases and Their Inhibitors in Human Platelets JUN-1CHI KAMBAYASHI AND MASATO SAKON 37. Measurement of Changes in Platelet Cyclic AMP in Vitro and in Vivo by Prelabeling Techniques: Application to the Detection and Assay of Circulating PGI2 RICHARD J. HASLAM AND MARIANNE VANDERWEL

355 371

386 415

430 442

455

VOLUME 170 NUCLEOSOMES

Section I. Chromatin and Nucleosomes: Preparative Techniques 1. Preparation of Nucleosomes and Chromatin ROGER D. KORNBERG, JAN W. LAPOINTE, AND YAHLI LORCH 2. Preparation of Simian Virus 40 Minichromosomes PIERRE OUDET, ETIENNE WEISS, AND EVELYNE REGNIER

3 14

CONTENTS OF VOLUMES 168-174, 176-194 3. Isolation of Yeast Plasmid Chromatin ANN DEAN, DAVID S. PEDERSON, AND ROBERT T. SIMPSON 4. Affinity Isolation of Replicating Simian Virus 40 Chromosomes TIMOTHY M. HERMAN

xv

26 41

S e c t i o n II. C h r o m a t i n a n d N u c l e o s o m e s : A n a l y t i c a l T e c h n i q u e s 5. Sucrose Gradient Techniques and Applications to Nucleosome Structure HANS NOLL AND MARKUSNOLL 6. Electrophoretic Analyses of Nucleosomes and Other Protein-DNA Complexes SuE-YING HUANG AND WILLIAM T. GARRARD 7. Electron Microscopy of Chromatin Jose M. SoGo AND FRITZ THOMA 8. Electron Microscopy of Simian Virus 40 Minichromosomes PIERRE OUDET, PATRICK SCHULTZ, JEAN-CLAUDE HOMO, AND PIERRE COLIN 9. Immunoelectron Microscopy of Nucleosomes CHRISTOPHER L. F. WOODCOCK 10. Use of Fluorescent Probes to Study Nucleosomes JOAN-RAMON DABAN AND CHARLES R. CANTOR l 1. Preparation and Application of Immunological Probes for Nucleosomes MICHAEL BUSTIN 12. Use of Antihistone Antibodies with Nucleosomes SYLVIANE MULLER AND MARC H. V. VAN REGENMORTEL 13. Digestion of Nucleosomes with Deoxyribonucleases I and 11 LEONARD C. LUTTER 14. Analysis of Hypersensitive Sites in Chromatin CARL WU 15. Application of Nucleases to Visualizing Chromatin Organization at Replication Forks MICHAEL E. CUSICK, PAUL M. WASSARMAN,AND MELVIN L. DEPAMPHILIS 16. Nuclease Digestion of Transcriptionally Active Chromatin MARIA BELLARD, GUY DRETZEN, ANGELA GIANGRANDE, AND PHILIPPE RAMAIN 17. Analysis of RNA Polymerase III Transcription in Vitro Using Chromatin and Cloned Gene Templates JOEL M. GOTTESEELD 18. Nonenzymatic Cleavage of Chromatin IAIN L. CARTWRIGHT AND SARAH C. R. ELGIN 19. Chemical Radiolabeling of Lysines That Interact Strongly with DNA in Chromatin JEAN O. THOMAS 20. Mapping DNA-Protein Interactions by Cross-Linking A. D. MIRZABEKOV, S. G. BAVYKIN, A. V. BELYAVSKY,V. L. KARPOV. O. V. PREOBRAZHENSKAYA,V. V. SHICK, AND K. K. EBRAL1DSE 21. Analysis of Nucleosome Positioning by Indirect End-Labeling and Molecular Cloning S . A . NEDOSPASOV, A. N. SHAKHOV, AND G. P. GEORGIEV 22. Crystallization of Nucleosome Core Particles D. RHODES, R. S. BROWN, AND A. KLUG S e c t i o n III.

55 116 142 166 180 192 214 "2-51 264 269

290

317 347 359 369

386 408 420

Histories: Preparation and Analysis

23. Isolation and Characterization of Histones C. VON HOLT, W. F. BRANDT, H. J. GREYLING, G. G. LINDSEY. J. D. RETIEF, J. DE A. RODRIGUES, 5. SCHWAGER, AND B. T. SEWELE

431

CONTENTS OF VOLUMES 168-174, 176-194

xvi

24. Purification and Analysis of HI Histones R. DAVIDCOLE 25. Analysis of Histone Subtypes and Their Modified Forms by Polyacrylamide Gel Electrophoresis RONALD W. LENNOXAND LEONARDH. COHEN 26. Chemical Cross-Linking of Histones JEAN O. THOMAS

524 532 549

S e c t i o n IV. A s s e m b l y a n d R e c o n s t i t u t i o n o f C h r o m a t i n 27. Assembly of Nucleosomes and Chromatin in Vitro DANIELARHODESAND RONALDA. LASKEY 28. Reconstitution of Chromatin from Purified Components ARNOLD STEIN 29. Assembly of Chromatin with Oocyte Extracts AKIKO SHIMAMURA,BRETJESSEE, AND ABRAHAMWORCEL 30. Purification of Xenopus Egg Nucleoplasmin and Its Use in Chromatin Assembly

575 585 603

in Vitro

LINDA SEALY, RICHARDR. BURGESS,MATT GOTTEN, AND ROGERCHALKLEY 31. Analysis of Nucleosome Positioning by in Vitro Reconstitution BERND NEUBAUERAND WOLFRAMHORZ

612 630

V O L U M E 171 BIOMEMBRANES PART R TRANSPORT THEORY: CELLS AND MODEL MEMBRANES

S e c t i o n I. T r a n s p o r t T h e o r y 1. Transport Machinery: An Overview GEORGESACHSAND SIDNEY FLEISCHER 2. Survey of Carrier Methodology: Strategy for Identification, Isolation, and Characterization of Transport Systems MARTIN KLINGENBERG 3. Kinetics of Transport: Analyzing, Testing, and Characterizing Models Using Kinetic Approaches WILFRED D. STEIN 4. Kinetics of Ion Movement Mediated by Carriers and Channels OLAF SPARREANDERSEN 5. Inhibition Kinetics of Carrier Systems ROSA DEVILSAND R. M. KRUPKA 6. Design of Simple Devices to Measure Solute Fluxes and Binding in Monolayer Cell Cultures JAYDUTTV. VADGAMA 7. Utilization of Binding Energy and Coupling Rules for Active Transport and Other Coupled Vectorial Processes WILLIAM P. JENCKS 8. Generation of Steady-State Rate Equations for Enzyme and Carrier-Transport Mechanisms: A Microcomputer Program KEITH R. RUNYANAND ROBERTB. GUNN

3 12 23 62 113 133 145

164

S e c t i o n II. M o d e l M e m b r a n e s a n d T h e i r C h a r a c t e r i s t i c s 9. Liposome Preparation and Size Characterization MARTIN C. WOODLEAND DEMETRIOSPAPAHADJOPOULOS 193 10. Preparation of Microcapsules from Human Erythrocytes: Use in Transport Experiments of Glutathione and Its S-Conjugate TAKAHITOKONDO 217

CONTENTS OF VOLUMES 168-174, 176-194

xvii

11. Planar Lipid-Protein Membranes: Strategies of Formation and of Detecting Dependencies of Ion Transport Functions on Membrane Conditions HANSGEORG SCHINDLER

225

12. Spontaneous Insertion of Integral Membrane Proteins into Preformed Unilamellar Vesicles

DAVID ZAK1M AND ANTHONY W. SCOTTO

253

13. Gentle and Fast Transmembrane Reconstitution of Membrane Proteins NORBERT A. DENCHER

14. Ion Carriers in Planar Bilayers: Relaxation Techniques and Noise Analysis R. BENZ, H.-A. KOL8, P. L~UGER, AND G. STARK 15. Ion Interactions at Membranous Polypeptide Sites Using Nuclear Magnetic Resonance: Determining Rate and Binding Constants and Site Locations

265

274

DAN W. URRY, T. L. TRAPANE, C. M. VENKATACHALAM, AND

R. B. McMICHENS

286

DAVID CAFISO, ALAN MCLAUGHLIN, STUART MCLAUGHLIN. AND ANTHONY WINISKI

342

16. Measuring Electrostatic Potentials Adjacent to Membranes

17. Determination of Surface Potential of Biological Membranes HAGAI ROTTENBERG

364

18. Lipid Coumarin Dye as a Probe of Interracial Electrical Potential in Biomembranes PETER FROMHERZ 19. Modulation of Membrane Protein Function by Surface Potential

376

REINHARD KR;~MER

387

S e c t i o n III.

Isolation of Cells

20. Use of Nonequilibrium Thermodynamics in the Analysis of Transport: General Flow-Force Relationships and the Linear Domain DANIELA PIETROBON AND S. ROY CAPLAN

397

21. Cell Isolation Techniques: Use of Enzymes and Chelators MIGUEL J. M. LEWIN AND A. M. CHERET

444

22. Cell Separation by Gradient Centrifugation Methods THOMAS G. PRETLOW AND THERESA P. PRETLOW

23. Cell Separation by Elutriation: Major and Minor Cell Types from Complex Tissues MARTIN J. SANDERS AND ANDREW H. SOLL 24. Cell Separation Using Velocity Sedimentation at Unit Gravity and Buoyant Density Centrifugation JOHN WELLS 25. Separation of Cell Populations by Free-Flow Electrophoresis

462

482 497

HANS-G. HEIDRICH AND KURT HANNIG

513

26. Separation of Cells and Cell Organelles by Partition in Aqueous Polymer TwoPhase Systems PER-AKE ALBERTSSON 27. Flow Cytometry: Rapid Isolation and Analysis of Single Cells

532

BRUCE D. JENSEN AND PAUt. KARL HORAN

549

28. Use of Cell-Specific Monoclonal Antibodies to Isolate Renal Epithelia ARLYN GARCIA-PEREZ, WILLIAM S. SPIELMAN, WILLIAM K. SONNENBURG, AND WILLIAM L. SMITH

581

29. Pancreatic Acini as Second Messenger Models in Exocrine Secretion JERRY D. GARDNER AND ROBERT T. JENSEN

59()

30. Ascites Cell Preparation: Strains, Caveats ROSE M. JOHNSTONE AND PHILIP C. LARIS

593

xviii

CONTENTS OF VOLUMES 168-174, 176-194

S e c t i o n IV. P o l a r C e l l S y s t e m s 31. Direct Current Electrical Measurement in Epithelia: Steady-State and Transient Analysis WARREN S. REHM, M. SCHWARTZ,AND G. CARRASQUER 32. Impedance Analysis in Tight Epithelia CHRIS CLAUSEN 33. Electrical Impedance Analysis of Leaky Epithelia: Theory, Techniques, and Leak Artifact Problems L . G . M . GORDON, G. KOTTRA,AND E. FROMTER 34. Patch-Clamp Experiments in Epithelia: Activation by Hormones or Neurotransmitters OLE H. PETERSEN 35. Ionic Permeation Mechanisms in Epithelia: Biionic Potentials, Dilution Potentials, Conductances, and Streaming Potentials PETER H. BARRY 36. Use of Ionophores in Epithelia: Characterizing Membrane Properties SIMON A. LEWIS AND NANCY K. WILLS

607 628 642 663 678 715

37. Cultures as Epithelial Models: Porous-Bottom Culture Dishes for Studying Transport and Differentiation JOSEPH S. HANDLER, NORDICA GREEN, AND RODERICK E. STEELE

736

38. Volume Regulation in Epithelia: Experimental Approaches ANTHONY D. C. MACKNIGHTAND JOHN P. LEADER

744

39. Scanning Electrode Localization of Transport Pathways in Epithelial Tissues J. KEVIN FOSKETT AND CARL SCHEFFEY

792

S e c t i o n V. M o d i f i c a t i o n o f Cells 40. Experimental Control of Intracellular Environment PETER F. BAKERAND DEREKE. KNIGHT 41. Implantation of Isolated Carriers and Receptors into Living Cells by Sendal Virus Envelope-Mediated Fusion

817

ABRAHAM LOYTER, N. CHEJANOVSKY, AND V. CITOVSKY

829

42. Resonance Energy Transfer Microscopy: Visual Colocalization of Fluorescent Lipid Probes in Liposomes PAUL S. USTER AND RICHARD E. PAGANO 43. Permeabilizing Mammalian Cells to Macromolecules

850

GARY A. WEISMAN, KEVIN D. LUSTIG, ILAN FRIEDBERG, AND LEON A. HEPPEL

857

V O L U M E 172 BIOMEMBRANES PART S TRANSPORT: MEMBRANE ISOLATION AND CHARACTERIZATION

S e c t i o n I. M e m b r a n e S e p a r a t i o n 1. Membrane Isolation: Strategy, Techniques, Markers E. KINNE-SAFFRANAND R. K. H. KINNE 2. Isolation of Plasma Membranes from Polar Cells and Tissues: Apical/Basolateral Separation, Purity, Function AUSTIN K. MIRCHEFF 3. Transport-Specific Fractionation for Purification of ATP-Dependent Ca 2+ Pumps STANLEY M. GOLDIN AND STEVEN C. KING

4. Regulation of Membrane Transport by Endocytotic Removal and Exocytotic Insertion of Transporters QAIS AL-AWQATI

3 18 34

49

CONTENTS OF VOLUMES 168-174, 1 7 6 - 1 9 4

S e c t i o n II. T r a n s p o r t

Methods

xix

in Cells and Vesicles

5. Proton Electrochemical Potential Gradient in Vesicles, Organelles, and Prokaryotic Cells HAGAI ROTTENBERG 6. Measurement of pH of Intracellular Compartments in Living Cells by Fluorescent Dyes JANET VAN ADELSBERG, JONATHAN BARASCH, AND QA1S AL-AWQATI

7. Use of Carbocyanine Dyes to Assay Membrane Potential of Mouse Ascites Tumor Cells A. ALAN EDDY 8. Optical Measurement of Membrane Potential in Cells, Organelles, and Vesicles J. C. FREEDMAN AND T. S. NOVAK 9. Fluorescence Methods for Continuous Monitoring of Transport in Cells and Vesicles OFER EIDEEMAN AND Z. IOAV CABANTCHIK 10. Preparation and Use of Micro- and Macroelectrodes for Measurement of Transmembrane Potentials and Ion Activities DANIEL AMMANN AND PICO CARONI

63

85

95 102 122

136

1I. Ion Channel-Mediated Fluxes in Membrane Vesicles: Selective Amplification of Isotope Uptake by Electrical Diffusion Potentials HAIM GARTY AND STEVEN J. D. KARLISH

12. Use of Calcium-Regulated Photoproteins as Intracellular Ca-'- Indicators JOHN R. BLINKS 13. Electron Probe X-ray Microanalysis of Ca -'+, Mg 2-, and Other Ions in Rapidly F r o z e n Cells A . V . SOMLYO, H. SHUMAN. AND A. P. SOMLYO 14. Measurement of Cytosolic Free Ca 2+ with Quin2

155

164 203

ROGER TSIEN AND TULLIO POZZAN

230

15. Analyzing Transport Kinetics with Desk-Top Hybrid Computers HAROLD G. HEMPLING 16. Synthesis and Properties of Caged Nucleotides

262

JEFFERY W. WALKER, GORDON P. REID, AND DAVID R. TRENTHAM

288

17. Measurement of Ion Fluxes in Membrane Vesicles Using Rapid-Reaction Methods HERBERT S. CHASE, JR., MARK U. GELERNT, AND MARC C. DEBELL 18. Tracer Studies with Isolated Membrane Vesicles ULRICH HOPFER 19. Electron Paramagnetic Resonance Methods for Measuring pH Gradients. Transmembrane Potentials, and Membrane Dynamics DAVID S. CAFISO 20. Transport Studies with Renal Proximal Tubular and Small Intestinal Brush Border and Basolateral Membrane Vesicles: Vesicle Heterogeneity, Coexistence of Transport Systems

301

3[3 331

HEINI MURER, PIOTR GMAJ, BRUNO STIEGER, AND BRUNO HAGENBUCH

346

21. Dynamic Laser Light Scattering to Determine Size Distributions of Vesicles HORST RUF, YANNIS GEORGAL1S, AND ERNS'I- GRELL

364

Section III. Membrane

Analysis and Characterization

22. Sensitive Protein Assay in Presence of High Levels of Lipid RONALD S. KAPLAN AND PETER L. PEDERSEN

23. Orienting Synthetic and Native Biological Membranes for Time-Averaged and Time-Resolved Structure Determinations L. G. HERBETTE AND J. K. BLASIE 24. Radiation Inactivation of Membrane Components and Molecular Mass Determination by Target Analysis E . S . KEMPNER AND SIDNEY FEEISCHER

393

399 410

CONTENTS OF VOLUMES 168-174, 176-194

XX

25. Prediction of Bilayer Spanning Domains of Hydrophobic and Amphipathic Membrane Proteins: Application to the Cytochrome b and Colicin Families J. W. SHIVER, A. A. PETERSON, W. R. WIDGER, P. N. FURBACHER, AND W. A. CRAMER

26. Order and Viscosity of Membranes: Analysis by Time-Resolved Fluorescence Depolarization MAARTEN P. HEYN 27. Rotational and Translational Diffusion in Membranes Measured by Fluorescence and Phosphorescence Methods THOMASM. JOVIN AND WINCHIL L. C. VAZ 28. Membrane Protein Molecular Weight Determined by Low-Angle Laser LightScattering Photometry Coupled with High-Performance Gel Chromatography YUTARO HAYASHI, HIDEO MATSUI, AND TOSHIO TAKAGI

439 462

471

514

29. Critical Micellar Concentrations of Detergents M. ZUEAUF, U. Ft0RSTENBERGER, M. GRABO, P. JAGGI, M. REGENASS, AND J. P. ROSENBUSCH

528

30. High-Performance Liquid Chromatography of Membrane Lipids: Glycosphingolipids and Phospholipids R. H. McCLUER, M. D. ULLMAN, AND F. B. JUNGALWALA 538 31. Circular Dichroism for Determining Secondary Structure and State of Aggregation of Membrane Proteins MAARTENP. HEYN 575 32. Cross-Linking Techniques H.G. BXUMERTAND H. FASOLD 584 33. Membrane-lmpermeant Cross-Linking Reagents JAMES V. STAROS AND P. S. R. ANJANEYULU

609

34. Photochemical Labeling of Apolar Phase of Membranes JOSEF BRUNNER 35. Electrophoretic Transfer of High-Molecular-Weight Proteins for Immunostaining

628

KUAN WANG, BRADFORD O. FANGER, CHERYL A. GUYER, AND JAMES V. STAROS

687

36. Size and Shape of Membrane Protein-Detergent Complexes: Hydrodynamic Studies

STEVEN CLARKE AND MURRAY D. SMIGEL

696

V O L U M E 173 BIOMEMBRANES PART T CELLULAR AND SUBCELLULAR TRANSPORT: EUKARYOTIC (NONEPITHELIAL) CELLS

S e c t i o n I. R e d B l o o d Cells

A. I n t a c t S y s t e m 1. Structure and Deformation Properties of Red Blood Cells: Concepts and Quantitative Methods EVAN A. EVANS 2. Ektacytometry of Red Blood Cells ROBERT M. JOHNSON 3. Methods and Analysis of Erythrocyte Anion Fluxes

3 35

ROBERT B. GUNN AND OTTO FROHLICH

54

4. Cation Fluxes in the Red Blood Cell: Na ÷, K ÷ Pump JOHN R. SACHS 5. Use of Triphenylmethylphosphonium to Measure Membrane Potentials in Red Blood Cells JEFFREY C. FREEDMANAND TERRI S. NOVAK

80 94

CONTENTS OF VOLUMES 168--174, 176--194 6. Measurement and Control of Intracellular Calcium in Intact Red Cells VIRG1LIO L. LEW AND JAVIER GARC[A-SANCHO 7. Recording Single-Channel Currents from Human Red Cells W. SCHWARZ, R. GRYGORCZYK, AND D. HOF 8. Identification of Amino Acid Transporters in the Red Blood Cell CATHERINE M. HARVEY AND J. CLIVE ELLORY 9. Transport Measurement of Anions, Nonelectrolytes, and Water in Red Blood Cell and Ghost Systems JESPER BRAHM 10. Kinetic Properties of Na+/H" Exchange and Li-/Na +. Na+/NaL and Na'/Li + Exchanges of Human Red Cells MITZY CANESSA ll. Water Channels across the Red Blood Cell and Other Biological Membranes ARTHUR K. SOLOMON 12. Transbilayer Mobility of Phosphatidylcholine in the Red Blood Cell Jos A. F. Oa DEN KAMP AND BEN ROELOFSEN 13. Sugar Transport in Red Blood Cells W . F . WIDDAS 14. Nucleoside Transport across Red Cell Membranes Z. 1. CABANTCHIK 15. Transport Studies in Red Blood Cells by Measuring Light Scattering GONTER FRED FUHRMANN 16. Cation-Anion Cotransport MARK HAAS AND THOMAS J. MCMANUS 17. Sodium-Calcium and Sodium-Proton Exchangers in Red Blood Cells JOHN C. PARKER 18. Monocarboxylate Transport in Red Blood Cells: Kinetics and Chemical Modification BERNHARD DEUTICKE 19. Alkali Metal/Proton Exchange PETERM. CALA AND KAREN S. HOFFMANN

xxi

[00 1[2 122 160 176 192 223 231 250 263 280 292 300 330

B. Derived Red Cell Preparations 20. Preparation of Resealable Membranes Maximally Depleted of Cytosolic Components by Gel Filtration and Tryptic Digestion: Resealable White Ghosts PHILLIP G. WOOD 21. Hemolytic Holes in Human Erythrocyte Membrane Ghosts MICHAEL R. LIEDER AND THEODORE L. STECK 22. Preparation and Properties of One-Step Inside-Out Vesicles from Red Cell Membranes JAV1ER GARC~A-SANCHOAND JAVIER ALVAREZ 23. Na*,K+-Pump Stoichiometry and Coupling in Inside-Out Vesicles from Red Blood Cell Membranes RHODA BLOSTEIN AND WILLIAM J. HARVEY 24. Preparation of Red Cell Membrane Skeleton Proteins WILLIAM C. HORNE, THOMAS L. LETO, AND RICHARD A. ANDERSON 25. Reconstitution of Red Blood Cells: Cytoskeletons Coated with Phospholipid Membrane CHAN Y. JUNG AND JOHN CUPPOLETTI

346 356 368 377 380 392

C. Anion Transporter 26. Impact of Methodology on Studies of Anion Transport: An Overview ASER ROTHSTEIN 27. Isolation, Reconstitution, and Assessment of Transmembrane Orientation of the Anion-Exchange Protein Z . I . CABANTCHIK 28. Proteolytic Cleavage of the Anion Transporter and Its Orientation in the Membrane MICHAEL J. A. TANNER 29. Functional Asymmetry of the Anion-Exchange Protein, Capnophorin: Effects on Substrate and Inhibitor Binding PHILIP A. KNAUF AND JESPER BRAHM

401 410 423 432

xxii

CONTENTS OF VOLUMES 168-174, 176-194

30. Measurement of Erythroid Band 3 Protein-Mediated Anion Transport in mRNAInjected Oocytes of Xenopus laevis R. GRYGORCZYK, P. HANKE-BAIER, W. SCHWARZ, AND H. PASSOW

453

31. Chemical Modification of the Anion-Transport System with Phenylglyoxal POUL J. BJERRUM

466

JOSEPH R. CASEY, DEBRA M. LIEBERMAN, AND REINHART A. F. REITHMEIER

494

32. Purification and Characterization of Band 3 Protein 33. Protein Associations with Band 3 at Cytoplasmic Surface of Human Erythrocyte Membrane

AHMAD WASEEM AND THEODORE L. STECK

S e c t i o n II. O t h e r M a m m a l i a n

513

Cells: Intact Cells

34. Transport of Glutathione, Glutathione Disulfide, and Glutathione Conjugates across the Hepatocyte Plasma Membrane THEO P. M. AKERBOOM AND HELMUT SIES 35. Ca + Fluxes and Phosphoinositides in Hepatocytes P. F. BLACKMORE AND J. H. EXTON 36. Magnesium Transport in Eukaryotic and Prokaryotic Cells Using Magnesium-28 Ion ROBERT D. GRUBBS, MARSHALL D. SNAVELY, S. PAUL HMIEL, AND MICHAEL E. MAGUIRE

37. Measurement of Amino Acid Transport by Hepatocytes in Suspension or Monolayer Culture MICHAEL S. KILBERG 38. Distinguishing Amino Acid Transport Systems of a Given Cell or Tissue

523

534

546

564

HALVOR N. CHRISTENSEN

576

39. Measuring Hexose Transport in Suspended Cells JORGEN GLIEMANN 40. Preparation and Culture of Embryonic and Neonatal Heart Muscle Cells: Modification of Transport Activity KARL WERDAN AND ERLAND ERDMANN 41. Isolation of Calcium-Tolerant Atrial and Ventricular Myocytes from Adult Rat Heart

616

MARY BETH DE YOUNG, BARTOLOMEOGIANNATTASIO,AND ANTONIO SCARPA

662

42. Measurement of Na + Pump in Isolated Cells LEIGH H. ENGLISH AND JOHN T. SCHULZ 43. Measurement of Na÷-K + Pump in Muscle RAYMOND A. SJOD|N 44. Measurement of Transport versus Metabolism in Cultured Cells ROBERT M. WOHLHUETER AND PETER G. W. PLAGEMANN

634

676

695 714

45. Purification and Reconstitution of the Phosphate Transporter from Rat Liver Mitochondria RONALD S. KAPLAN, RAYMOND D. PRATT, AND PETER L. PEDERSEN

46. Measurement of Vacuolar pH and Cytoplasmic Calcium in Living Cells Using Fluorescence Microscopy FREDERICK R. MAXFIELD 47. Transport in Mouse Ascites Tumor Cells: Symport of Na ÷ with Amino Acids A. ALAN EDDY AND E. R. JOHNSON 48. Measurements of Cytoplasmic pH and Cellular Volume for Detection of Na+/H ÷ Exchange in Lymphocytes S. GRINSTE1N, S. COHEN, J. D. GOETZ-SMITH, AND S. J. DIXON

732

745 771

777

CONTENTS OF VOLUMES 168-174, 176-194

xxiii

VOLUME 174 BIOMEMBRANES PART U

C E L L U L A R AND SUBCELLULAR TRANSPORT:

EUKARYOTIC (NoNEPITHEL1AL) CELLS

Section I. Transport in Subcellular Organelles of Animals

A. Plasma Membranes and Derived Transporters 1. Measurement of Transported Calcium in Synaptosomes DAVIDG. NICHOLLS 2. Glutamate Accumulation into Synaptic Vesicles PHILL1P E. KISH AND TETSUFUMI UEDA 3. Identification of Bile Acid Transport Protein in Hepatocyte Sinusoidal Plasma Membranes DANIEL LEVY AND PATRICIA VON DIPPE 4. Transport of Alanine across Hepatocyte Plasma Membranes JOHN D. McGIVAN 5. Purification and Reconstitution of Glucose Transporter from Human Erythrocytes STEPHEN A. BALDWIN AND GUSTAV E. LIENHARD 6. Isolation of Bilitranslocase, the Anion Transporter from Liver Plasma Membrane for Bilirubin and Other Organic Anions GIAN LUIGI SOTTOCASA, GIAN CARLO LUNAZZI, AND CLAUDIO TIRIBEI_LI

3 9 25 31 39

50

B. Intracellular Organelles 7. Measurement of Intactness of Rat Liver Endoplasmic Reticulum WILLIAM J. ARION 8. Calcium Ion Transport in Mitochondria NILS-ERIK LEO SARIS AND ASHLEY ALLSHIRE 9. Measurement of Proton Leakage across Mitochondrial Inner Membranes and Its Relation to Protonmotive Force DAVIDG. NICHOLLS AND EDUARDO RIAL 10. Influence of Calcium Ions on Mammalian Intramitochondrial Dehydrogenases JAMES G. McCORMACKAND RICHARD M. DENTON ll. Effects of Hormones on Mitochondrial Processes SIBYLLE SOBOLL AND HELMUT SIES 12. Use of Fluorescein Isothiocyanate-Dextran to Measure Proton Pumping in Lysosomes and Related Organelles SHOJI OHKUMA 13. Cystine Exodus from Lysosomes: Cystinosis MARGARET L. SMITH, ALICE A. GREENE, JERRY A. SCHNEIDER, RONALD L. PISONI, AND HALVOR N. CHRISTENSEN 14. Isolation of Physiologically Responsive Secretory Granules from Exocrine Tissues ULRICH HOPFER AND KENNETH GASSER 15. Transport of Nucleotides in the Golgi Complex BECCA FLEISCHER

58 68 85 95 ll8 131

154 162 173

CONTENTS OF VOLUMES 168-174, 176-194

xxiv

S e c t i o n II. T r a n s p o r t in P l a n t s

A. H i g h e r P l a n t s 16. Water Flow in Plants and Its Coupling to Other Processes: An Overview ERNST STEUDLE

183

OK YOUNG LEE-STADELMANN AND EDUARD J. STADELMANN 18. Passive Permeability EDUARD J. STADELMANNAND

225

OK YOUNGLEE-STADELMANN 19. Compartmentation in Root Cells and Tissues: X-Ray Microanalysis of Specific

246

17. Plasmolysis and Deplasmolysis

Ions

ANDRt~ L~UCHLI AND PATRICKJ. BOURSIER

267

A.H. DE BOER

277

GABRIELE ORLICH AND EWALD KOMOR

288

20. Xylem Transport 21. Phloem Transport

22. Patch Clamp Measurements on Isolated Guard Cell Protoplasts and Vacuoles KLAUS RASCHKE AND RAINER HEDRICH

312

23. Intracellular and Intercellular pH Measurement with Microelectrodes D. J. F. BOWLING

331

U. ZIMMERMANN

338

Transport Systems in Algae and Bryophytes: An Overview J.A. RAVEN Uptake of Sugars and Amino Acids by Chlorella W. TANNERAND N. SAUER Electrophysiology of Giant Algal Cells M.J. BEILBY Ion Transport in Chara Cells WILLIAM J. LUCASAND DALE SANDERS Light-Induced Hyperpolarization in Nitella ROGER M. SPANSWlCK ATP-Driven Chloride Pump in Giant Alga Acetabularia D. GRADMANN

366 390 403 443 479 490

24. Water Relations of Plant Cells: Pressure Probe Technique B. L o w e r P l a n t s 25. 26. 27. 28. 29. 30.

C. O r g a n e l l e s 31. Transport in Isolated Yeast Vacuoles: Characterization of Arginine Permease THOMAS BOLLER, MATHIAS DURR, AND ANDRES WlEMKEN

504

32. Metabolite Levels in Specific Cells and Subcellular Compartments of Plant Leaves MARK STITT, ROSS McC.LILLEY, RICHARD GERHARDT, AND HANS W. HELDT

518

33. Isolation of Plant Vacuoles and Measurement of Transport EWALD KOMOR AND MARGARETTHOM

552

S e c t i o n III. T r a n s p o r t in S i n g l e - C e l l E u k a r y o t e s : F u n g a l Ceils 34. Kinetic Studies of Transport in Yeast ARNOgT KOTYK 567 35. Proton Extrusion in Yeast ARNOgT KOTYK 592 36. Ion Transport in Yeast Including Lipophilic Ions GEORGE W. F. H. BORST-PAUWELS 603 37. Sugar Transport in Normal and Mutant Yeast Cells VINCENT P. CIRILLO 617 38. Transport of Amino Acids and Selected Anions in Yeast A. ALAN EDDY AND PAMELA HOPKINS

623

39. Accumulation of Electroneutral and Charged Carbohydrates by Proton Cotransport in Rhodotorula MILAN HOFER

629

CONTENTS OF VOLUMES 168--174, 141--194 40. Proton-Potassium Symport in Walled Eukaryotes: Neurospora CLIFFORD L. SLAYMAN AND GERALD N. ZUCKIER 41. Isolation of Everted Plasma Membrane Vesicles from Neurospora crassa and Measurement of Transport Function GENE A. SCARBOROUGH

XXV

654 667

V O L U M E 176 N U C L E A R MAGNETIC RESONANCE PART A SPECTRAL TECHNIQUES AND DYNAMICS

S e c t i o n I. B a s i c T e c h n i q u e s I. One-Dimensional Nuclear Overhauser Effects and Peak Intensity Measurements JAMES A. FERRETTI AND GEORGE H. WEISS 2. Two-Dimensional Nuclear Magnetic Resonance Spectroscopy of Proteins: An Overview JOHN L. MARKLEY 3. Solvent Suppression P . J . HORE 4. Sample Preparation NORMAN J. OPPENHEIMER

3 12 64 78

S e c t i o n II. A d v a n c e d T e c h n i q u e s 5. Heteronuclear Nuclear Magnetic Resonance Experiments for Studies of Protein Conformation GERHARD WAGNER 6. Multiple-Quantum Nuclear Magnetic Resonance MARK RANCE, WALTER J. CHAZIN, CLAUDIO DALVIT, AND PETER E. WRIGHT 7. Detection of Insensitive Nuclei AD BAX, STEVEN W. SPARKS, AND DENNIS A. TORCHIA 8. Homonuclear Hartmann-Hahn Experiments AD BAX 9. Two-Dimensional Nuclear Overhauser Effect: Complete Relaxation Matrix Analysis BRANDAN A. BORGIAS AND THOMAS L. JAMES 10. Selective Relaxation Techniques NERI NICCOEAI AND CLAUDIO ROSSI II. Rotating-Frame Nuclear Overhauser Effect L. R. BROWN AND B. T. FARMER II 12. Modern Spectrum Analysis in Nuclear Magnetic Resonance: Alternatives to the Fourier Transform JEFFREY C. HOCH 13. Solid-State Nuclear Magnetic Resonance Structural Studies of Proteins S. J. OPELLA AND P. L. STEWART

93 114 134 151 169 184 199 216 242

Section III. Enzyme Dynamics A. Rate Constants 14. Nuclear Magnetic Resonance Line-Shape Analysis and Determination of Exchange Rates B . D . NAGESWARARAO 15. Applicability of Magnetization Transfer Nuclear Magnetic Resonance to Study Chemical Exchange Reactions JENS J. LED, HENRIK GESMAR, AND FRITS ABILDGAARD

279

311

xxvi

CONTENTS OF VOLUMES 168-174, 176-194

16. Two-Dimensional Nuclear Magnetic Resonance Studies of Enzyme Kinetics and Metabolites in Vivo BRUCE A. BERKOWITZAND ROBERTS. BALABAN 17. Isotope Exchange PAUL ROSCH

330 342

B. M o l e c u l a r M o t i o n s 18. Interpreting Protein Dynamics with Nuclear Magnetic Resonance Relaxation Measurements ROBERT E. LONDON 19. Solid-State Deuterium Nuclear Magnetic Resonance Spectroscopy of Proteins MAX A. KENIRY 20. Protein Rotational Correlation Times by Carbon-13 Rotating-Frame Spin-Lattice Relaxation in Presence of Off-Resonance Radiofrequency Field THOMAS SCHLEICH, COURTNEYF. MORGAN,AND G. HERBERTCAINES 21. Measurement of Translational Motion by Pulse-Gradient Spin-Echo Nuclear Magnetic Resonance RONALD L. HANERAND THOMASSCHLEICH 22. Structural Characterization of Protein Folding Intermediates by Proton Magnetic Resonance and Hydrogen Exchange HEINRICH RODER

358 376

386 418 446

Appendix Computer Programs Related to Nuclear Magnetic Resonance: Availability, Summaries, and Critiques Addendum to Article [16] BRUCEA. BERI,:OWITZAND ROBERTS. BALABAN

477 493

V O L U M E 177 NUCLEAR MAGNETIC RESONANCE PART B STRUCTURE AND MECHANISM

S e c t i o n I. E n z y m e M o d i f i c a t i o n s for N u c l e a r M a g n e t i c Resonance Studies 1. Fluorine Nuclear Magnetic Resonance of Fluorinated Ligands J.T. GERIG 2. Deuteration in Protein Proton Magnetic Resonance DAVID M. LEMASTER 3. Expression and Nitrogen-15 Labeling of Proteins for Proton and Nitrogen-15 Nuclear Magnetic Resonance DAVID C. MUCHMORE,LAWRENCEP. MCINTOSH, CHRISTOPHER B. RUSSELL, D. ERIC ANDERSON,AND FREDERICKW. DAHLQUIST 4. Isotopic Labeling with Hydrogen-2 and Carbon-13 to Compare Conformations of Proteins and Mutants Generated by Site-Directed Mutagenesis, I DAVID W. HIBLER, LYNN HARPOLD, MARKDELL'ACQUA, TAYEBEH POURMOTABBED,JOHN A. GERLT, JOYCEA. WILDE, AND PHILIP H. BOLTON 5. Spin Labeling of Proteins PHYLLIS A. KOSEN

3 23

44

74 86

CONTENTS OF VOLUMES 168-174, 176-194

xxvii

Section II. Protein Structure 6. Determination of Three-Dimensional Protein Structures in Solution by Nuclear Magnetic Resonance: An Overview KURT WOTHRICH 7. Proton Nuclear Magnetic Resonance Assignments VLADIMIR J. BASUS 8. Computer-Assisted Resonance Assignments MARTIN BILLETER 9. Distance Geometry I . D . KUNTZ, J. F. THOMASON, AND C. M. OSHIRO 10. Molecular Dynamics Simulation Techniques for Determination of Molecular Structures from Nuclear Magnetic Resonance Data R. M. SCHEEK, W. F. VAN GUNSTEREN, AND R. KAPTEIN ll. Heuristic Refinement Method for Determination of Solution Structure of Proteins from Nuclear Magnetic Resonance Data Russ B. ALTMAN AND OEEG JARDETZKY 12. Proton Magnetic Resonance of Paramagnetic Metalloproteins IVANO BERTINI, LUCIA BANCI, AND CLAUDIO LUCHINAT 13. Phosphorus-31 Nuclear Magnetic Resonance of Phosphoproteins HANS J. VOGEL 14. Isotopic Labeling with Hydrogen-2 and Carbon-13 to Compare Conformations of Proteins and Mutants Generated by Site-Directed Mutagenesis, I1 JOYCE A. WILDE, PHILIP H. BOLTON, DAVID W. HIBLER, LYNN HARPOLO, TAYEBEH POURMOTABBED, MARK DELL'ACQUA, AND JOHN A. GERLT

S e c t i o n III.

Enzyme

125 132 150 159

204

218 246 263

282

Mechanisms

15. Phosphorus-31 Nuclear Magnetic Resonance of Enzyme Complexes: Bound Ligand Structure, Dynamics, and Environment DAVID G. GORENSTEIN 16. Ligand-Protein Interactions via Nuclear Magnetic Resonance of Quadrupolar Nuclei CHARLES R. SANDERS II AND MING-DAW TSA1 17. Ligand Conformations and Ligand-Enzyme Interactions as Studied by the Nuclear Overhauser Effect PAUL R. ROSEVEAR AND ALBERT S. MILDVAN 18. Determination of Equilibrium Constants of Enzyme-Bound Reactants and Products by Nuclear Magnetic Resonance B . D . NAGESWARA RAO 19. Mechanistic Studies Utilizing Oxygen-18 Analyzed by Carbon-13 and Nitrogen15 Nuclear Magnetic Resonance Spectroscopy JOHN M. RISLEY AND ROBERT L. VAN ETTEN 20. Positional Isotope Exchange Using Phosphorus-31 Nuclear Magnetic Resonance JOSEPH J. VILLAFRANCA 21. Paramagnetic Probes of Macromolecules JOSEPH J. VIELAFRANCA

S e c t i o n I V . I n Vivo S t u d i e s o f E n z y m a t i c

295 317 333 358

376 390 403

Activity

22. Enzyme Regulation of Metabolic Flux SHEILA M. COHEN 23. Monitoring Intracellular Metabolism by Nuclear Magnetic Resonance JACK S. COHEN, ROBBE C. LYON, AND PETER F. DALY

417 435

Appendix Computer Programs Related to Nuclear Magnetic Resonance: Availability, Summaries, and Critiques

455

xxviii

CONTENTS OF VOLUMES 168-174, 176-194 VOLUME 178 ANTIBODIES, A N T I G E N S , AND MOLECULAR MIMICRY

S e c t i o n I. I d i o t y p e s , A n t i - i d i o t y p e s , a n d M o l e c u l a r M i m i c r y

A. Concepts and Methods 1. Idiotypic Networks and Nature of Molecular Mimicry: An Overview HEINZ KOHLER, SRIN1VASKAVERI, THOMAS KIEBER-EMMONS, W. JOHN W. MORROW, SYBILLE MULLER, AND SYAMALRAYCHAUDHURI 2. Production and Characterization of Anti-idiotypic Antibody Reagents MICHAEL R. SCHICK AND RONALD C. KENNEDY 3. Production and Characterization of Monoclonal Idiotypes and Anti-idiotypes for Small Ligands PAUL H. KuSSIE, GEORGE ALBRIGHT, AND D. SCOTT LINTHICUM 4. Use of Peptides Encoded by Complementary RNA for Generating Anti-idiotypic Antibodies of Predefined Specificity J. EDWIN BLALOCK, JOHN N. WHITAKER, ETTY N. BENVENISTE,AND KENNETH L. BOST 5. Serological Methods to Detect Anti-idiotypic Antibodies F. PEROSA, T. KAGESHITA,R. ONO, AND S. FERRONE 6. Molecular Analysis of Monoclonal Idiotypes and Anti-idiotypes PAUL H. KUSSIE, MARK A. SHERMAN, DARIO MARCHETTI, AND D. SCOTT LINTHICUM 7. Size-Exclusion High-Performance Liquid Chromatography in Analysis of Protein and Peptide Epitopes FRED J. STEVENS 8. Immunoelectron Microscopy of idiotype-Anti-idiotype Complexes KENNETH H. ROUX 9. Use of Immunoblotting to Detect Idiotypic Determinants on Monoclonal Antibodies C. PETIT, M. E. SAURON, M. GILBERT, AND J. THI~ZE 10. Idiotopes Carried by Antiallergen Antibodies MARC G. JACQUEMINAND JEAN-MARIE R. SA1NT-REMY l l . Structure-Function Analysis of Protein Active Sites with Anti-idiotypic Antibody DAVID S. LUDWIG AND GARY K. SCHOOLNIK 12. Competitive Idiotype-Anti-idiotype Enzyme Immunoassay for Adenosine Deaminase Binding Protein MITCHELL J. NELLES

3 36

49

63 74

91 107 130 144 150 163 171

B. A n t i - i d i o t y p e M o l e c u l a r M i m i c r y a n d R e c e p t o r s 13. Interaction of Anti-idiotypic Antibodies with Membrane Receptors: Practical Considerations A . D . STROSBERG 14. Anti-idiotypic Antibodies as Probes of Hormone Receptor Structure and Function NADIR R. FARID 15. Immunocytochemistry of Receptors Using Anti-idiotypic Antibodies KARL M. KNIGGE, DIANE T. PIEKUT, LEO G. ABOOD, SHIRLEY A. JOSEPH, GREGORY J. MICHAEL, LU XIN, AND DAVID J. BERLOVE 16. Production and Properties of Antimorphine Anti-idiotypic Antibodies and Their Antiopiate Receptor Activity JAY A. GLASEL

179 191

212 222

CONTENTS OF VOLUMES 168-174, 176-194 17. Production and Characterization of Anti-idiotypic Antiopioid Receptor Antibodies CHRISTIAN GRAMSCH, RUDIGER SCHULZ. SABIN KOSIN. AHMED H. S. HASSAN, AND ALBERT HERZ 18. Anti-idiotypic Antibodies That Interact with/3-Adrenergic Catecholamine Receptor A . D . STROSBERG 19. Characterization and Properties of Antisubstance P Anti-idiotypic Antibodies JEAN-YVES COURAUD, SYLVIE MAILLET, JACQUES GRASSI, YVELINE FROBERT, AND PHILIPPE PRADELLES 20. Production and Properties of Anti-idiotypic Antibodies That Recognize Insulin Receptor IRUN R. COHEN, DANA ELIAS, MICHA RAPOPORT, AND YORAM SHECHTER 21. Anti-idiotypic Antibodies as Probes of Prolactin Receptor MOUSSA B. H. YOUDIM, RONNIE J. BARKEY, T. AMIT, AND J. GUy 22. Development and Use of Antireceptor Antibodies to Study Interaction of Mammalian Reovirus Type 3 with Its Cell Surface Receptor WILLIAM V. WILLIAMS, DAVID B. WEINER, AND MARK 1. GREENE

xxix

243 265

275

300 308

321

C. Anti-idiotype Vaccines and Immunotoxins 23. Use of Anti-idiotypic Antibodies to Treat Lymphoid Tumors GEORGE T. STEVENSON 24. Preparation and Use of Anti-idiotypic Antibodies Armed with Holotoxins or Hemitoxins in Treatment of B-Cell Neoplasms SANDRA BRIDGES, DAN L. LONGO, AND RICHARD J. YOULE 25. Use of Anti-idiotypic Antibody-Drug Conjugates to Treat Experimental Murine B-Cell Tumors E. HURWITZ AND J. HAIMOVICH 26. Monoclonal Anti-idiotypic Antibody Vaccines against Poliovirus, Canine Parvovirus, and Rabies Virus G. F. RIMMELZWAAN, E. J. BUNSCHOTEN, F. G. C. M. UYTDEHAAG, AND A. D. M. E. OSTERHAUS 27. Use of Anti-idiotypic Antibodies in Approaches to Immunoprophylaxis of Schistosomiasis JEAN-MARIE GRZYCH, FLORENCE ROUSSEE-VELGE, AND ANDRI~ CAPRON 28. Selective Immunotoxins Prepared with Mutant Diphtheria Toxins Coupled to Monoclonal Antibodies MARCO COLOMBATTI, LORENA DELL'ARC1PRETE, RINO RAPPUOLI, AND GIUSEPPE TRIDENTE 29. Immune Suppression of Anti-DNA Antibody Production Using Anti-idiotypic Antibody-Neocarzinostatin Conjugates TAKESHI SASAKI, YOSHIO KO1DE, AND KAORU YOSHIGAGA 30. Anti-idiotypic Antibodies in the Study and Treatment of Experimental Autoimmune Myasthenia Gravis MIRIAM C. SOUROUJON AND SARA FUCHS 31. Inhibition of Autoimmune Reactivity against Acetylcholine Receptor with ldiotype-Specific Immunotoxins KEITH A. KROLICK S e c t i o n II. E n g i n e e r e d

341

356 369

375

390

404

422 433 448

Antibodies

32. Production and Properties of Chimeric Antibody Molecules SEUNG-UON SHIN AND SHERIE L. MORRISON 33. Expression of Engineered Antibodies and Antibody Fragments in Microorganisms MARC BETTER AND ARNOLD H. HORWITZ

459 476

CONTENTS OF VOLUMES 168-174, 176-194

XXX

34. Expression of Functional Antibody Fv and Fab Fragments in Escherichia coli ANDREAS PLOCKTHUN AND ARNE SKERRA

497

35. Recombinant Antibodies Possessing Novel Effector Functions TED W. LOVE, MARSCHALL S. RUNGE, EDGAR HABER, AND THOMAS QUERTERMOUS

515

S e c t i o n III. C a t a l y t i c A n t i b o d i e s or A b z y m e s 36. Production of Antibodies That Mimic Enzyme Catalytic Activity ALFONSO TRAMONTANO AND DIANE SCHLOEDER

531

37. Design of Catalytic Antibodies SCOTT J. POLLACK, GRACE R. NAKAYAMA,AND PETER G. SCHULTZ

551

S e c t i o n IV. E p i t o p e D e s i g n a n d A p p l i c a t i o n s 38. Use of Hydrophilicity Plotting Procedures to Identify Protein Antigenic Segments and Other Interaction Sites THOMAS P. HOPI" 39. Computer Prediction of B-Cell Determinants from Protein Amino Acid Sequences Based on Incidence of fl Turns VIKTOR KRCHlqAK, OTAKAR MACH, AND ANTONfN MAL¢~

571

586

40. Identification of T-Cell Epitopes and Use in Construction of Synthetic Vaccines JAMES L. CORNETTE, HANAH MARGALIT, CHARLES DELISI, AND

JAY A. BERZOFSKY 41. Use of Synthetic T-Cell Epitopes as Immunogens to Induce Antibodies to Hepatitis B Components DAVID R. MILICH AND GEORGE B. THORNTON 42. Peptide Vaccines Based on Enhanced Immunogenicity of Peptide Epitopes Presented with T-Cell Determinants or Hepatitis B Core Protein

611 634

MICHAEL J. FRANCIS AND BERWYN E. CLARKE

659

43. Production and Properties of Site-Specific Antibodies to Synthetic Peptide Antigens Related to Potential Cell Surface Receptor Sites for Rhinovirus J. McCKAY AND G. WERNER

676

44. Custom-Designed Synthetic Peptide Immunoassays for Distinguishing HIV Type 1 and Type 2 Infections JOHN W. GNANN, JR., LAURA L. SMITH, AND MICHAEL B. A. OLDSTONE

693

45. Production and Use of Synthetic Peptide Antibodies to Map Region Associated with Sodium Channel Inactivation HAMUTAL MEIRI, MAREI SAMMAR, AND ARIELA SCHWARTZ

714

46. Multiple Antigenic Peptide Method for Producing Antipeptide Site-Specific Antibodies

DAVID N. POSNETT AND JAMES P. TAM

739

47. Protein Footprinting Method for Studying Antigen-Antibody Interactions and Epitope Mapping

HOOSHMANDSHESHBERADARANAND LENDON G. PAYNE

746

CONTENTS OF VOLUMES 168-174, 176-194

xxxi

V O L U M E 179 COMPLEX CARBOHYDRATES PART F

S e c t i o n I. A n a l y t i c a l M e t h o d s 1. Characterization of Human Tracheobronchial Mucin Glycoproteins MARY CALLAGHAN ROSE

3

2. Characterization of O-Linked Saccharides from Cell Surface Glycoproteins MINORU FUKUDA

17

3. Lectin Affinity Chromatography of Glycolipids and Glycolipid-Derived Oligosaccharides

DAVID F. SMITH AND B. VIRGINIA TORRES

4. Affinity Chromatography of Oligosaccharides on E4-PhytohemagglutininAgarose Column AKIRA KOBATAAND KATSUKOYAMASHITA 5. Analysis and Purification of Oligosaccharides by High-Performance Liquid Affinity Chromatography D. ZOPF, S. OHLSON, J. DAKOUR,W. WANG,AND A. LUNDBLAD 6. Separation of Oligosaccharides Using High-Performance Anion-Exchange Chromatography with Pulsed Amperometric Detection R. REID TOWNSEND, MARK R. HARDY, AND YVAN CHUAN LEE

30

46

55

65

7. Monosaccharide Analysis of Glycoconjugates by High-Performance Anion-Exchange Chromatography with Pulsed Amperometric Detection MARK R. HARDY

76

SIDNEY W. WHITEHEART, ANTONINO PASSANITI, JONATHAN S. REICHNER. GORDON D. HOLT, ROBERT S. HALT1WANGER,AND GERALD W. HART

82

8. Glycosyltransferase Probes

9. Immunoblottingand Immunobindingof Acidic Polysaccharides Separated by Gel Electrophoresis GRADIMIRN. MISEVIC 10. Polyacrylamide Gel Electrophoresis of Capsular Polysaccharides of Bacteria

95

SINIKKA PELKONEN AND JUKKA FINNE

104

11. Carbohydrate Receptor Binding Using Colloidal Gold VICTORIA KOLB-BACHOFEN 12. Two-Dimensional Proton Magnetic Resonance Spectroscopy

1I I

JANUSZ DABROWSKI

122

ROGER A. LAINE

157

13. Tandem Mass Spectrometry of Oligosaccharides

Section II. P r e p a r a t i o n s 14. Glycosphingolipids: Structure, Biological Source, and Properties CHERYL L. M. STULTS, CHARLES C. SWEELEY, AND BRUCE A. MACHER 15. Mycobacterial Glycolipids: Isolation, Structures, Antigenicity, and Synthesis of Neoantigens MICHAEL MCNEIL, DELPHI CHATTERJEE, SHIRLEY Wu HUNTER, AND PATRICK J. BRENNAN

167

215

16. Synthesis and Characterization of Ganglioside Gm Derivatives: Lyso-GM3, DeN-acetyl-GM3, and Other Compounds GUSTAVO A. NORES, NOBUO HANAI, STEVEN B. LEVERY, HUGH L. EATON, MARY ELLEN K. SALYAN, AND SEN-ITIROH HAKOMORI

242

xxxii

CONTENTS OF VOLUMES 168--174, 176-194

17. Conjugation of Glycopeptides to Proteins YUANC. LEE AND REIKO T. LEE 18. Conversion of Galactosyl Residues to 6-Amino Derivatives for Attachment of Functional Groups REIKO T. LEE AND YUAN C. LEE 19. Quantitative Conversion of Mucin-Type Sugar Chains to Radioactive Oligosaccharides JUNKO AMANO AND AKIRA KOBATA 20. Specific Labeling of Cell Surface Poly-N-acetyllactosamine Glycans DOROTHE SPILLMANN AND JUKKA FINNE 21. Preparation of Cytidine 5'-Monophospho-N-Acetylneuraminic Acid and Uridine 5'-Diphosphoglucuronic Acid; Synthesis of a-2,6-Sialyllactosamine, a-2,6Sialyllactose, and Hyaluronic Acid ETHAN S. SIMON, ERIC J. TOONE, GARY OSTROFF, MARK D. BEDNARSKI, AND GEORGE M. WHITESIDES

Section III. Carbohydrate-Binding

253 257 261 270

275

Proteins

22. Human Monoclonal Antibodies Associated with Neuropathy RICHARD H. QUARLES 23. Production of Carbohydrate-Specific Human Monoclonal Antibodies in Vitro LESLIE n. SILBERSTEIN, LEIGH C. JEFFERIES, JUNE GOLDMAN, AND STEVEN L. SPITALNIK 24. Phosphomannosyl Receptors from Bovine Testis MAOMI LI, JACK J. DISTLER, AND GEORGE W. JOURDIAN 25. Mannose/N-Acetylglucosamine-Binding Proteins from Mammalian Sera TOSHISUKE KAWASAKI, NOBUKO KAWASAKI, AND IKUO YAMASHINA 26. Galactose/N-Acetylgalactosamine-Binding Protein and Mannose/L-Fucose/NAcetylglucosamine-Binding Proteins from Rat Peritoneal Macrophages TOSHISUKE KAWASAKI, MASAYUKi h, AND IKUO YAMASHINA 27. Snowdrop Lectin HANAE KAKU AND IRWIN J. GOLDSTEIN 28. Allomyrina dichotoma Lectins KATSUKO YAMASHITA, AKIRA KOBATA, TSUNEO SUZUKI, AND KAZUO UMETSU 29. Vegetative Tissue Lectins from Dolichos biflorus MARILYNNE. ETZLER

291

299 304 310

322 327

331 341

Section IV. Biosynthesis 30. High-Performance Liquid Chromatography Assays for N-Acetylglucosaminyltransferascs Involved in N- and O-Glycan Synthesis HARRY SCHACHTER, INKA BROCKHAUSEN, AND ERIC HULL 31. Glycosyltransferase Assays Using Pyridylaminated Acceptors: N-Acetylglucosaminyltransferase III, IV, and V NAOYUKI TANIGUCHI, ATSUSHI NISHIKAWA, SHIGERU FUJII, AND JIANGUO GU 32. Sialate 9-O-Acetylesterase from Rat Liver HERMAN H. HIGA, ADRIANA MANZI, SANORA DIAZ, AND AJIT MARK! 33. Glycoprotein Sialate 7(9)-O-Acetyltransferase from Rat Liver Golgi Vesicles SANDRA DIAZ, HERMAN H. HIGA, AND AJIT VARKI

351

397 409 416

CONTENTS OF VOLUMES 168-174, 1 7 6 - 1 9 4

xxxiii

34. Microsomal Sulfation of Proteochondroitin, Chondroitin, and Chondroitin Oligosaccharides GEETHA SUGUMARAN, DONALD E. HUMPHRIES, AND JEREMIAH E. SILBERT

422

35. Decreasing Sulfation of Proteoglycans Produced by Cultured Cells DONALD E. HUMPHRIES, GEETHA SUGUMARAN, AND JEREMIAH E. SILBERT

428

36. Specific Activity of Radiolabeled Hexosamines in Metabolic Labeling Experiments MASAKI YANAGISHITA, ANTONIETTA SALUSTRI, AND VINCENT C. HASCALL

435

37. Mannosidase IA from Rat Liver Golgi Membranes DAULAT RAM P. TULSIANI AND OSCAR TOUSTER

446

38. Glycoprotein Processing Enzymes of Plants GUR P. KAUSHAL AND ALAN D. ELBEIN

452

S e c t i o n V. D e g r a d a t i o n 39. Ceramide Glycanase from Leech, Hirudo medicinalis, and Earthworm, Lumbricus terrestris Yu-TEH LI AND Su-CHEN Ll 40. Endoglycoceramidase from Rhodococcus Species G-74-2

479

MAKOTO ITO AND TATSUYA YAMAGATA

488

41. Endo-fl-galactosidase Releasing Gal (al---~3)Gal 42. Lysosomal a-D-Mannosidase from Rat Epididymis

TAKASm MURAMATSU

496

DAULAT RAM P. TULSIANI AND OSCAR TOUSTER

500

43. Endoglycosidases from Flavobacterium meningosepticum Application to Biological Problems STEPHEN ALEXANDER AND JOHN H. ELDER

505

Section VI. Miscellaneous 44. Gangliosides That Modulate Membrane Protein Function YASUYUKI IGARASHI, HISAO NOJIRI, NOBUO HANAI. AND SEN-ITIROH HAKOMORI

521

45. Adhesion of Eukaryotic Cells to Immobilized Carbohydrates RONALD L. SCHNAAR, BRIAN K. BRANDLEY, LEILA K. NEEDHAM, PATTI SWANK-HILL, AND CAROL C. BLACKBURN

542

46. Glycolipid Transfer Protein from Pig Brain TERUKATSU SASAKI AND AKIRA ABE

559

47. Di- and Trigalacturonic Acid and A4'5-Di- and A4,5-Trigalacturonic Acids: Inducers of Proteinase Inhibitor Genes in Plants THOMAS MOLOSHOK AND CLARENCE A. RYAN

566

48. Glycation of Copper/Zinc Superoxide Dismutase and Its Inactivation: Identification of Glycated Sites NAOYUKI TANIGUCHI, KATSURA ARAI, AND NORIAKI KINOSHITA

5'70

xxxiv

CONTENTS OF VOLUMES

168-174, 1 7 6 - 1 9 4

V O L U M E 180 R N A PROCESSING PART A GENERAL METHODS

S e c t i o n I. P r e p a r a t i o n of S u b s t r a t e s

A. P u r i f i c a t i o n o f R N A M o l e c u l e s 1. Isolation of RNA SHELBY L. BERGERAND JOHN M. CHIRGWlN 2. Purification of RNA Molecules by Gel Techniques TOSHIMICHIIKEMURA 3. Purifying RNA by Column Chromatography N. KYLE TANNER

3 14 25

B. I n Vitro S y n t h e s i s o f R N A 4. SynthesisofLong, Capped Transcripts in VitrobySP6andT7RNAPolymerases JOEL K. YISRAELIAND DOUGA. MELTON 5. Synthesis of Small RNAs Using T7 RNA Polymerase JOHN F. MILLIGAN AND OLKE C. UHLENBECK

42 51

6. In Vitro Synthesis of End-Mature, Intron-Containing Transfer RNAs VICENTE M. REYESAND JOHN N. ABELSON

63

C. I n V i v o P r e p a r a t i o n o f S u b s t r a t e s 7. Preparation of Precursors to mRNA from Mammalian Cell Nuclei SELINA CHEN-KIANG AND DANIEL J. LAVERY

69

8. Pulse Labeling of Heterogeneous Nuclear RNA in Isolated Nuclei SELINA CHEN-KIANG AND DANIEL J. LAVERY

82

9. Isolation and Characterization of Yeast Ribosomal RNA Precursors and Preribosomes JACOBUS KLOOTWIJK AND RUDI J. PLANTA 10. Preparation of Yeast Transfer RNA Precursors in Vivo GAYLE KNAPP

96 ll0

S e c t i o n II. C h a r a c t e r i z a t i o n o f R N A s

A. P r i m a r y S t r u c t u r e 11. Dideoxy Sequencing of RNA Using Reverse Transcriptase CHANG S. HAHN, ELLEN G. ST~USS, AND JAMESH. ST~USS 12. RNA Fingerprinting

121

ANDREA D. BRANCH, BONNIE J. BENENFELD, AND HUGH D. ROBERTSON

130

13. Enzymatic RNA Sequencing Y. KUCHINOAND S. NISHIMURA 14. Characterization of Cap Structures YASUHIROFURUICH!AND AARONJ. SHATKIN 15. Isolation and Characterization of Branched Oligonucleotides from RNA J. DAVID REILLY, JOHN C. WALLACE, RANDA F. MELHEM, DAVID W. KOPP, AND MARY EDMONDS

154 164

177

CONTENTS OF VOLUMES 168-174, 176-194

xxxv

B. Secondary Structure 16. Enzymatic Approaches to Probing of RNA Secondary and Tertiary Structure GAYLE KNAPP 17. A Guide for Probing Native Small Nuclear RNA and Ribonucleoprotein Structures ALAIN KROL AND PHILIPPE CARBON 18. Phylogenetic Comparative Analysis of RNA Secondary Structure BRYAN D. JAMES, GARY J. OLSEN, AND NORMAN R. PACE 19. Mapping the Genetic Organization of RNA by Electron Microscopy Louise T. CHOW AND THOMAS R. BROKER 20. Computer Prediction of RNA Structure MICHAEL ZUKER 21. RNA Pseudoknots: Structure, Detection, and Prediction C. W. A. PLEIJ AND L. BOSCH 22. Absorbance Melting Curves of RNA JOSEPH D. PUGLIS1 AND IGNAClO TINOCO, JR.

192 212 227 239 262 289 304

C. RNA Functions 23. Nuclease Digestion: A Method for Mapping Introns SHELBY L. BERGER 24. Characterization of RNA Molecules by SI Nuclease Analysis ARNOLD J. BERK 25. Primer Extension Analysis of RNA WILLIAM R. BOORSTEIN AND ELIZABETH A. CRAIG 26. Modification Interference Analysis of Reactions Using RNA Substrates LAURA CONWAY AND MARVIN WICKENS

325 334 347 369

Section III. R N A Interactions A. Cross-Linking 27. Photoaffinity Cross-Linking Methods for Studying RNA-Protein Interactions MICHELLE M. HANNA 28. Ultraviolet-Induced Cross-Linking of RNA to Proteins in Vivo SERAFfN PIIqoL-ROMA, STEPHEN m. ADAM, YANG DO CHOI, AND GIDEON DREYFUSS 29. Analysis of Ultraviolet-lnduced RNA-RNA Cross-Links: A Means for Probing RNA Structure-Function Relationships ANDREA D. BRANCH, BONNIE J. BENENFELD, CYNTHIA P. PAUL, AND HUGH D. ROBERTSON

383

410

418

B. Other Methods 30. Analysis of Splicing Complexes and Small Nuclear Ribonucleoprotein Particles by Native Gel Electrophoresis MARIA M. KONARSKA 31. Determination of RNA-Protein and RNA-Ribonucleoprotein Interactions by Nuclease Probing KATHERINE A. PARKERAND JOAN A. STEITZ 32. Immunoprecipitation of Ribonucleoproteins Using Autoantibodies JOAN A. STEITZ

442 454 468

Xxxvi

CONTENTS OF VOLUMES 168-174, 176-194

33. Electron Microscopy of Ribonucleoprotein Complexes on Nascent RNA Using Miller Chromatin Spreading Method YVONNE N. OSHEIM AND ANN L. BEYER 34. Genetic Methods for Identification and Characterization of RNA-RNA and RNA-Protein Interactions ROY PARKER

481 510

Section IV. Appendix 35. Compilation of Small Nuclear RNA Sequences 36. Sequences and Classification of Group I and Group II Introns

RAM REDDY

521

JOHN M. BURKE 37. Compilation of Self-Cleaving Sequences from Plant Virus Satellite RNAs and Other Sources GEORGE BRUENING

533 546

VOLUME 181 RNA PROCESSINGPART B SPECIFIC METHODS

Section I. Processing of Messenger RNAs A. Mammalian Cell Extracts and Assays 1. Preparation and Fractionation of Mammalian Extracts Active in Pre-mRNA Splicing ANGELA KR,~MER AND WALTER KELLER 2. Small-Scale Preparation of Extracts from Radiolabeled Cells Efficient in PremRNA Splicing KEVIN A. W. LEE AND MICHAEL R. GREEN 3. Preparation of Active Nuclei WILLIAM F. MARZLUFF 4. Nonaqueous Isolation of Transcriptionally Active Nuclei from Xenopus Oocytes ELSEBET LUND AND PHILIP L. PAINE 5. Transient Transfection of HeLa Cells for Analysis of Splicing Reaction MARKUS AEBI 6. Preparation of Mammalian Extracts Active in Polyadenylation CLAIRE L. MOORE 7. Histone-Specific RNA 3' Processing in Nuclear Extracts from Mammalian Cells CLAUDIA STAUBER, DOMINIQUE SOLDATI, BERNHARD LOSCHER, AND DANIEL SCHUMPERLI

3 20 30 36 43 49

74

B. Yeast Extracts and Assays 8. Preparation and Fractionation of Yeast Splicing Extract Soo-CHEN CHENG, ANDREW NEWMAN, REN-JANG LIN, GLORIA D. MCFARLAND, AND JOHN N. ABELSON 9. Affinity Chromatography with Biotinylated RNAs S. W. RUBY, S. E. GOELZ, Z. HOSTOMSKY, AND J. N. ABELSON 10. Measurement and Analysis of Yeast Pre-mRNA Sequence Contribution to Splicing Efficiency B. C. RYMOND, C. PIKIELNY, B. SERAPHIN, P. LEGRAIN, AND M. ROSBASH

89 97

122

CONTENTS OF VOLUMES 168-174, 176-194

xxxvii

C. Extracts and Assays from Other Systems ll. Analysis of Pre-mRNA Processing in Transfected Plant Protoplasts GREGORY J. GOODALL, KARIN WIEBAUER, AND WITOLD FILIPOWICZ

148

D. Purified Enzymes 12. Polyadenylate Polymerases MARY EDMONDS 13. Purification and Use of Vaccinia Virus Messenger RNA Capping Enzyme STEWART SHUMAN AND BERNARD MOSS 14. RNA Lariat Debranching Enzyme as Tool for Analyzing RNA Structure BARBARA RUSKIN AND MICHAEL R. GREEN 15. Escherichia coli Ribonuclease III HUGH D. ROBERTSON 16. Messenger RNA Turnover in Cell-Free Extracts GARY BREWER AND JEFFREY ROSS 17. Nuclear Cap Binding Protein from HeLa Cells MUTSUHITO OHNO AND YOSHIRO SHIMURA

161 170 180 189 202

209

E. Small Nuclear Ribonucleoproteins 18. Purification of Small Nuclear Ribonucleoprotein Particles Active in RNA Processing ANGELA KRAMER 19. Purification of Small Nuclear Ribonucleoprotein Particles with Antibodies against Modified Nucleosides of Small Nuclear RNAs MONTSERRAT BACH, PETER BRINGMANN, AND REINHARD L(IHRMANN 20. Isolation of Small Nuclear Ribonucleoprotein Polypeptides SAELIEO. HOCH 21. Purification and Characterization of U Small Nuclear Ribonucleoproteins in Cesium Chloride Gradients CLAUDE BRUNEL AND GUY CATHALA 22. Structural Analysis of U Small Nuclear Ribonucleoproteins by in Vitro Assembly JORG HAMM AND IAIN W. MATTAJ 23. Site-Specific Degradation of RNA of Small Nuclear Ribonucleoprotein Particles with Complementary Oligodeoxynucleotides and RNase H ANGELA KR,~.MER

215

232 257 264 273

284

F. Heterogeneous Nuclear Ribonucleoproteins 24. Isolation and in Vitro Assembly of Nuclear Ribonucleoprotein Particles and Purification of Core Particle Proteins STANLEY F. BARNETT, STEPHANIE J, NORTHINGTON, AND WALLACE M. LESTOURGEON 25. Physical Methods for Characterization of Heterogeneous Nuclear Ribonucleoprotein Complexes JERZY T. SCHONEICH AND JOHN O. THOMAS 26. Immunological Methods for Purification and Characterization of Heterogeneous Nuclear Ribonucleoprotein Particles SERAFfN PI/qOL-ROMA, YANG DO CHOI, AND GIDEON DREYFUSS 27. Purification and Characterization of Proteins of Heterogeneous Nuclear Ribonucleoprotein Complexes by Affinity Chromatography SERAFfN PllqOL-ROMA, MAURICE S. SWANSON, MICHAEL J. MATUNIS, AND GIDEON DREYFUSS 28. Purification and Characterization of Polyadenylate-Binding Protein ALAN B. SACHS AND ROGER D. KORNBERG

293 307

317

326 332

xxxviii

CONTENTS OF VOLUMES 168-174, 176-194

S e c t i o n II. P r o c e s s i n g o f R i b o s o m a l R N A s 29. Preparation of Extracts and Assay of Ribosomal RNA Maturation in Escherichia coli

ANAND K. SRIVASTAVAAND DAVID SCHLESSINGER

355

30. Ribosomal RNA Terminal Maturase: Ribonuclease M5 from Bacillus subtilis NORMAN R. PACE AND BERNADETTE PACE

366

Section III. Processing of Transfer RNAs

A. Genetics 31. Yeast Suppressor Mutations and Transfer RNA Processing MARK NICHOLS, IAN WILLIS, AND DIETER SOLL

377

32. Genetics of Transfer RNA Processing in Escherichia coli YOSHIRO SHIMURA AND HIDEAKI SHIRAISHI

395

33. Genetic Methods for Study of Trans-Acting Genes Involved in Processing of Precursors to Yeast Cytoplasmic Transfer RNAs ANITA K. HOPPER

400

B. E n z y m o l o g y

of Processing

34. Ribonucleases Active at 3' Terminus of Transfer RNA MURRAY P. DEUTSCHER 35. Transfer RNA Nucleotidyltransferase MURRAY P. DEUTSCHER 36. Yeast Extracts for Transfer RNA Gene Transcription and Processing

421 434

CLAIRE F. EVANS AND DAVID R. ENGELKE

439

37. Enzymatic Addition of Guanylate to Histidine Transfer RNA JACINTA B. WILLIAMS, LYNN COOLEY, AND DIETER SOLL

38. Purification of Yeast Transfer RNA Ligase QI Xu, ERIC M. PHIZICKY, CHRIS L. GREER, AND JOHN N. ABELSON 39. Highly Purified Transfer RNA Splicing Endonuclease from Saccharomyces cereoisiae PHILLIP R. GREEN AND JOHN N. ABELSON 40. Purification and Characterization of Wheat Germ RNA Ligase and Associated Activities

L E S L I EPICK, HENRY M. FURNEAUX, AND JERARD HURWITZ

451

463 471 480

41. RNA 3'-Terminal Phosphate Cyclase from HeLa Cells WITOLD FILIPOWICZ AND OSCAR VICENTE

499

42. Transfer RNA Splicing Endonuclease from Xenopus laevis D. GANDINI-ATTARDI, I. M. BALDI, E. MATTOCCIA, AND G. P. TOCCHINI-VALENTINI

510

S e c t i o n IV. R i b o z y m e s 43. Genetic and Molecular Analysis of RNA Splicing in Escherichia coli MARLENE BELFORT, KAREN EHRENMAN, AND P. SCOTT CHANDRY 44. Genetic Analysis of RNA Splicing in Yeast Mitochondria

521

PHILIP S.. PERLMAN

539

45. Self-Splicing and Enzymatic Cleavage of RNA by a Group I Intervening Sequence JOHN A. LATHAM, ARTHUR J. ZAUG, AND THOMASR. CECH

558

xxxix

CONTENTS OF VOLUMES 168--174, 176--194

46. Preparation and Characterization of RNase P from Escherichia coli MADELINE F. BAER, JOHN G. ARNEZ, CECILIA GUERRIER-TAKADA, AGUSTIN VIOQUE, AND SIDNEY ALTMAN 47. Characterization of Self-Cleavage of Viroid and Virusoid RNAs ANTHONY C. FORSTER, CHRISTOPHER DAVIES, CHERYL J. HUTCHINS, AND ROBERT H. SYMONS

569

583

VOLUME 182 G U I D E TO PROTEIN PURIFtCATION

1. Why Purify Enzymes?

ARTHUR KORNBERG

I

Section I. Developing Purification Procedures 2. Strategies and Considerations for Protein Purifications

STUART LINN

Section II. General Methods for Handling Proteins and Enzymes 3. Setting Up a Laboratory 4. Buffers: Principles and Practice 5. 6. 7. 8.

MURRAY P. DEUTSCHER

19

VINCENT S. STOLL AND JOHN S. BLANCHARD Measurement of Enzyme Activity EDWARD F. ROSSOMANDO Quantitation of Protein CHRISTA M. STOSCHECK Concentration of Proteins and Removal of Solutes THOMAS POHL Maintaining Protein Stability MURRAY P. DEUTSCHER

24 38 50 68 83

S e c t i o n III. O p t i m i z a t i o n o f S t a r t i n g M a t e r i a l s 9. Overproduction of Proteins in Escherichia coil: Vectors, Hosts, and Strategies AslS DAS 10. Overexpression of Proteins in Eukaryotes MARGARET K. BRADLEY II. Secretion of Heterologous Proteins in Escherichia coli I. BARRY HOLLAND, BRENDAN KENNY, BORIS STEIPE, AND ANDREAS PLOCKTHUN

93 112

132

Section IV. Preparation of Extracts and Subcellular Fractionation 12. Preparation of Extracts from Prokaryotes MILLARD CULL AND CHARLES S. MCHENRY 13. Preparation of Extracts from Yeast S. MICHAL JAZWINSKI

147 154

xl

CONTENTS OF VOLUMES 168-174, 176-194

14. Preparation of Extracts from Plants PETER GEGENHEIMER 15. Preparation of Extracts from Higher Eukaryotes JOHN DAVIDDIGNAM 16. Isolation of Subcellular Organelles BRIAN STORRIEAND EDWARDA. MADDEN 17. Preparation of Membrane Fractions JURIS OZOLS

174 194 203 225

S e c t i o n V. S o l u b i l i z a t i o n P r o c e d u r e s 18. Detergents: An Overview 19. Solubilization of Native Membrane Proteins 20. Solubilization of Protein Aggregates

JUDITH M. NEUGEBAUER 239 LEONARD M. HJELMELAND 253

FIONA A. O. MARSTON AND DONNA L. HARTLEY

21. Removal of Detergents from Membrane Proteins

LEONARDM. HJELMELAND

264

277

S e c t i o n VI. P u r i f i c a t i o n P r o c e d u r e s : B u l k M e t h o d s 22. Precipitation Techniques SASHA ENGLARD AND SAM SEIFTER 23. Precipitation of Proteins with Polyethylene Glycol KENNETHC. INGHAM

285 301

S e c t i o n VII. P u r i f i c a t i o n P r o c e d u r e s : C h r o m a t o g r a p h i c M e t h o d s 24. 25. 26. 27. 28. 29. 30.

Ion-Exchange Chromatography EDWARD F. ROSSOMANDO Gel Filtration EARLE STELLWAGEN Protein Chromatography on Hydroxyapatite Columns MARINAJ. GORBUNOEE Hydrophobic Chromatography ROBERT M. KENNEDY Chromatography on Immobilized Reactive Dyes EARLE STELLWAGEN Affinity Chromatography: General Methods STEVEN OSTROVE Affinity Chromatography: Specialized Techniques

309 317 329 339 343 357

STEVEN OSTROVE AND SHELLY WEISS

371

31. Chromatofocusing LALLAN GIRl 32. High-Performance Liquid Chromatography: Effective Protein Purification by

380

Various Chromatographic Modes ROMAN M. CHICZ AND FRED E. REGNIER

392

Section VIII. Purification Procedures: Electrophoretic Methods 33. One-Dimensional Gel Electrophoresis DAVID E. GARFIN 34. Protein Analysis Using High-Resolution Two-Dimensional Polyacrylamide Gel Electrophoresis

425

BONNIE S. DUNBAR, HITOMI KIMURA, A N D THERESE M. TIMMONS

441

35. Isoelectric Focusing 36. Gel-Staining Techniques 37. Elution of Protein from Gels

DAVlD E. GARFIN CARL R. MERRIL MICHAEL G. HARRINGTON

459 477 488

CONTENTS OF VOLUMES 168-174, 176-194

xli

S e c t i o n IX. S p e c i a l i z e d P u r i f i c a t i o n P r o c e d u r e s 38. Purification of Membrane Proteins THOMASC. THOMASAND MARKG. MCNAMEE 39. Purification of DNA-Binding Proteins by Site-Specific DNA Affinity Chromatography SHLOMO EISENBERG, STEPHEN C. FRANCESCONI, CHRIS CIVALIER, AND SCOTT S. WALKER

40. Purification of Glycoproteins 41. Purification of Multienzyme Complexes

499

521

CRAIG GERARD

529

PAUL A. SRERE AND CHRISTOPHER K. MATHEW'S

539

S e c t i o n X. C h a r a c t e r i z a t i o n of P u r i f i e d P r o t e i n s 42. Determination of Purity DAVID G. RHODESAND THOMASM. LAUE 43. Determination of Size, Molecular Weight, and Presence of Subunits

555

THOMAS M. LAUE AND DAVID G. RHODES

566

44. 45. 46. 47.

Amino Acid Analysis JURIS OZOLS 587 Limited N-Terminal Sequence Analysis PAUL MATSUDA1RA 602 Peptide Mapping RALPH C. JUDD 613 Analysis for Protein Modifications and Nonprotein Cofactors SAM SE1FTER AND SASHA ENGLARD DAVID OLLIS AND STEPHEN WHITE

48. Protein Crystallization

626 646

S e c t i o n XI. I m m u n o l o g i c a l P r o c e d u r e s 49. Preparation of Polyclonal Antibodies BONNIE S. DUNBAR AND ERIC D. SCHWOEBEL

663

50. Preparation of Monoclonal Antibodies BONNIE S. DUNBAR AND SHERI M. SKINNER

670

51. Protein Blotting and Immunodetection THERESE U . TIMMONS AND BONNIE S. DUNBAR

679

52. lmmunoprecipitation of Proteins 53. lmmunoassays

GARY L. FIRESTONE AND SANDRA U. WINGUTH CHARLES W. PARKER

688 700

Section XII. Additional Techniques 54. Radiolabeling of Proteins 55. Using Purified Protein to Clone Its Gene 56. Computer Analysis of Protein Structure

CHARLES W. PARKER JOHN M. WOZNEV PATRICK ARGOS

721 738 751

Section XIII. Concluding Remarks 57. Rethinking Your Purification Procedure

MURRAY P. DEUTSCHER

779

xlii

CONTENTS OF VOLUMES 168-174, 1 7 6 - 1 9 4

Appendix: Personal Perspectives on E n z y m e Purification 58. The Private Life of DNA Polymerase I ARTHURKORNBERG 59. Purification and Crystallization of Transaldolase from Candida utilis B. L. HORECKERAND O. TSOLAS 60. Discovery of Glutamine Synthetase Cascade EARL R. STADTMAN 61. Discovery, Resolution, Purification, and Function of Elongation Factors

783

KIVIE MOLDAVE

809

"]88

793

V O L U M E 183 MOLECULAR EVOLUTION: COMPUTER ANALYSIS OF PROTEIN AND NUCLEIC ACID SEQUENCES

S e c t i o n I. D a t a b a s e s 1. GenBank: Current Status and Future Directions CHRISTIANBURKSET AL. 2. EMBL Data Library PATRICIA KAHN AND GRAHAMCAMERON 3. Protein Sequence Database WINONA C. BARKER, DAVID G. GEORGE, AND LOIS T. HUNT

4. Cooperation between Databases and Scientific Community

BOR1VOJKEIL

3 23 31

50

S e c t i o n II. S e a r c h i n g D a t a b a s e s 5. Rapid and Sensitive Sequence Comparison with FASTP and FASTA WILLIAM R. PEARSON

63

6. Searching through Sequence Databases RUSSELL F. DOOLITTLE 7. Finding Protein Similarities with Nucleotide Sequence Databases

99

STEVEN HENIKOFF, JAMES C. WALLACE, AND JOSEPH P. BROWN

111

8. Use of Homology Domains in Sequence Similarity Detection CHARLES B. LAWRENCE

133

MICHAEL GgIRSKOV, ROLAND LCITHY, AND DAVID EISENBERG

146

9. Profile Analysis

S e c t i o n III. P a t t e r n s in N u c l e i c A c i d S e q u e n c e s 10. Finding Protein Coding Regions in Genomic Sequences RODGERSTADEN 11. Ancient Patterns in Nucleic Acid Sequences JOHN C. W. SHEPHERD 12. Searching for Patterns in Protein and Nucleic Acid Sequences RODGER STADEV4 13. Consensus Patterns in DNA GARY D. STORMO 14. Consensus Methods for DNA and Protein Sequence Alignment MICHAEL S. WATERMANAND ROBERT JONES

163 180 193 211 221

15. k-Tuple Frequency Analysis: From Intron/Exon Discrimination to T-Cell Epitope Mapping JEAN-MICHEL CLAVERIE, ISABELLE SAUVAGET, AND LYDIE BOUGUELERET

237

CONTENTS OF VOLUMES 168-174, 176-194

16. Splice Junctions, Branch Point Sites, and Exons: Sequence Statistics, Identification, and Applications to Genome Project PERIANNAN SENAI'ATHY,MARVINB. SHAPIRO,AND NOMI L. HARRIS

xliii

252

S e c t i o n IV. P r e d i c t i n g R N A S e c o n d a r y S t r u c t u r e s 17. Predicting Optimal and Suboptimal Secondary Structure for RNA JOHN h . JAEGER, DOUGLAS H. TURNER, AND MICHAEL ZUKER

281

18. Detecting Pseudoknots and Other Local Base-Pairing Structures in RNA Sequences HUGO M. MARTINEZ 19. Computer Modeling and Display of RNA Secondary and Tertiary Structures

306

DANIEL GAUTHERET, FRANCOIS MAJOR, AND ROBERT CEDERGREN

318

S e c t i o n V. A l i g n i n g P r o t e i n a n d Nucleic Acid S e q u e n c e s 20. Mutation Data Matrix and Its Uses DAVID G. GEORGE, WINONAC. BARKER,AND LOIS T. HUNT 21. Sensitivity Comparison of Protein Amino Acid Sequences

333

PATRICK ARGOS AND MARTIN VINGRON

352

22. Three-Way Needleman-Wunsch Algorithm MITSUOMURATA 23. Progressive Alignment and Phylogenetic Tree Construction of Protein Sequences

365

DA-FEI FENG AND RUSSELL F. DOOLITTLE

375

24. Identification of Significant Sequence Patterns in Proteins SAMUEL KARLIN, B. EDWIN BLAISDELL, AND VOLKER BRENDEL

388

25. Protein Multiple Sequence Alignment and Flexible Pattern Matching GEOFFREY J. BARTON

403

26. Genomic Divergence through Gene Rearrangement DAVID SANKOFF, ROBERT CEDERGREN, AND YVON ABEL

428

27. Multiple Sequence Comparison DAVIDJ. BACONAND WAYNEF. ANDERSON 28. Simultaneous Comparison of Several Sequences MAUNO VIHINEN 29. Hierarchical Method to Align Large Numbers of Biological Sequences

438 447

WILLIAM R. TAYLOR

456

JOHN F. COLLINS AND ANDREW F. W. COULSON

474

30. Significance of Protein Sequence Similarities 31. Fast Alignment of DNA and Protein Sequences GAD M. LANDAU, UZI VISHKIN, AND RUTH NUSSINOV

487

S e c t i o n VI. E s t i m a t i n g S e q u e n c e D i v e r g e n c e 32. Statistical Geometry on Sequence Space MANFRED EIGEN AND RUTHILD WINKLER-OSWATITSCH

505

33. Statistical Methods for Estimating Sequence Divergence TAKASHI GOJOBORI, ETSUKO N. MORIYAMA, AND MOTOO KIMURA

531

34. Converting Distance to Time: Application to Human Evolution HIROHISA KISHINO AND MASAM1 HASEGAWA

5.50

xliv

CONTENTS OF VOLUMES 168-174, 176-194

35. Influence of Base Composition on Quantitative Estimates of Gene Evolution CECILIA SACCONE,CECILIALANAVE, GRAZIANOPESOLE, AND GIULIANOPREPARATA 36. Maximum Likelihood Methods NARUYASA1TOU

570 584

S e c t i o n VII. P h y l o g e n e t i c T r e e s 37. Maximum Parsimony Approach_ to Construction of Evolutionary Trees from Aligned Homologous Sequences JOHN CZELUSNIAK, MORRIS GOODMAN, NANCY D. MONCRIEF, AND

SUZANNE M. KEHOE 38. Phylogeny Determination Using Dynamically Weighted Parsimony Method PATRICK L. WILLIAMSAND WALTERM. FITCH 39. Unified Approach to Alignment and Phylogenies JOTUN HEIN 40. Statistical Tests of Molecular Phylogenies WEN-HSIUNG Li AND MANOLO GOUY 41. Nearest Neighbor Procedure for Relating Progressively Aligned Amino Acid Sequences

RUSSELL F. DOOLITTLE AND DA-FEI FENG

601 615 626 645 659

42. Phylogenetic Relationships from Three-Dimensional Protein Structures MARK S. JOHNSON, ANDREJ ~ALI, AND TOM L. BLUNDELL

670

V O L U M E 184 AVIDIN-BIOTIN TECHNOLOGY

S e c t i o n I. I n t r o d u c t i o n 1. Reflections 2. Introduction to Avidin-Biotin Technology

FREDERIC M. RICHARDS

3

MEIR WILCHEK AND EDWARD A. BAYER

5

3. Applications of Avidin-Biotin Technology: Literature Survey MEIR WILCHEK AND EDWARD A. BAYER

14

S e c t i o n II. B i o t i n - B i n d i n g P r o t e i n s 4. Biotin-Binding Proteins: Overview and Prospects EDWARD A. BAYER AND MEIR WILCHEK

49

N. MICHAELGREEN

51

YAFFA HILLER, EDWARD A. BAYER, AND MEIR WILCHEK

68

5. Avidin and Streptavidin 6. Nonglycosylated Avidin 7. Cloning and Expression of Avidin in Escherichia coli

GYAN CHANDRA AND JOHN G. GRAY

70

8. Isolation and Properties of Streptavidin EDWARD A. BAYER, HAYA BEN-HUR, AND MEIR WILCHEK

9. Crystal Forms of Avidin 10. Nonavidin Biotin-Binding Proteins

80

ODED LIVNAHAND JOEL L. SUSSMAN

90

KRISHNAMURTI DAKSHINAMURTIAND JASBIR CHAUHAN

93

CONTENTS OF VOLUMES 168-174, 176-194 11. Biotinidase

BARRY WOLF, JEANNE HYMES, AND GREGORY S. HEARD

xlv 103

12. Monoclonal Antibody to Biotin KRISHNAMURTI DAKSHINAMURTI AND EDWARD S. RECTOR

I 11

Section III. G e n e r a l M e t h o d o l o g y

A. P r e p a r a t i o n o f B i o t i n D e r i v a t i v e s 13. Biotin-Containing Reagents

MEIR WILCHEKAND EDWARDA. BAYER

/23

14. Protein Biotinylation

EDWARD A. BAYER AND MEIR WILCHEK

138

15. Enzymatic C-Terminal Biotinylation of Proteins ALEXANDER SCHWARZ, CHRISTIAN WANDREY, EDWARD A. BAYER, AND MEIR WILCHEK

16. Antibodies Biotinylated via Sugar Moieties DANIEL J. O'SHANNESSY 17. 2-Iminobiotin-Containing Reagent and Affinity Columns BARBARA FUDEM-GOLDIN AND GEORGE A. ORR

160

162 167

B. P r e p a r a t i o n o f A v i d i n a n d S t r e p t a v i d i n D e r i v a t i v e s 18. Avidin- and Streptavidin-Containing Probes EDWARD A. BAYERAND MEIR WILCHEK 19. Phycobiliprotein-Avidinand Phycobiliprotein-Biotin Conjugates

174

ALEXANDER N. GLAZER AND LUBERT STRYER

188

20. Monovalent Avidin Affinity Columns RONALD A. KOHANSKI AND M. DANIEL LANE

194

21. Prevention of Nonspecific Binding of Avidin RAYMOND C. DUHAMEL AND JAMES S. WHITEHEAD

201

C. A s s a y s for A v i d i n a n d B i o t i n 22. Enzymatic and Radioactive Assays for Biotin, Avidin, and Streptavidin ERNEST V. GROMAN, JEFFREY M. ROTHENBERG. EDWARD A. BAYER, AND MEIR WILCHEK

23. Colorimetric Enzyme Assays for Avidin and Biotin EDWARD A. BAYER, HAYABEN-HUR, AND MEIR WILCHEK 24. Sequential Solid-Phase Assay for Biotin Based on ~251-LabeledAvidin DONALD M. MOCK 25. Fluorometric Assay for Avidin-Biotin Interaction DONALD M. MOCK AND PAUL HOROWITZ

208

217 224 234

S e c t i o n IV. Applications

A. I s o l a t i o n a n d P u r i f i c a t i o n 26. Isolation of Biologically Active Compounds: A Universal Approach MEIR WILCHEK AND EDWARD A. BAYER

243

27. Isolation and Characterization of Hormone Receptors FRANCES M. FINN AND KLAUS HOFMANN

244

xlvi

CONTENTS OF VOLUMES

168-174, 176-194

28. Biotinylation of Peptide Hormones: Structural Analysis and Application to Flow Cytometry WALTER NEWMAN, L. DAWSON BEALL, AND ZAFAR I. RANDHAWA 29. Purification of Gonadotropin-Releasing Hormone Receptors ELI HAZUM 30. Biotinylestradiol for Purification of Estrogen Receptor GI~.RARD REDEUILH, CLAUDE SECCO, AND ETIENNE-EMILE BAULIEU 31. One-Step Immunoaffinity Purification of Transferrin EDWARD A. BAYER AND MEIR WILCHEK 32. Lectin-Mediated Isolation of Cell Surface Glycoproteins GEOFFREY M. W. COOK AND J. WILLIAM BUCKLE 33. Immunoselective Cell Separation JAN WORMMEESTER, FRANK STIEKEMA, AND CORNEEIS DE GROOT 34. Isolation and Analysis of Splicing Complexes PAULA J. GRABOWSKI 35. Z-DNA Affinity Chromatography RICHARD FISHEL, PAUL ANZIANO, AND ALEXANDER RICH

275 285 292 301 304 314 319 328

B. Localization 36. Affinity Cytochemistry MEIR WILCHEK, EDWARD A. BAYER, AND EHUD SKUTELSKY 37. Double Fluorescence Immunolabeling Studies ANNE H. DUTTON, MARGIE ADAMS, AND S. J. SINGER 38. Detection of Cell Surface Components Using Fluorescent Microspheres ARTHUR J. SYTKOWSKI 39. Immunohistochemistry Su-MING Hsu 40. Immunocytochemical Detection of Human T- and B-Cell Antigens ROGER A. WARNKE AND LAWRENCE M. WEISS 41. Ultrastructural Localization of Retinal Photoreceptor Proteins DAVID S. PAPERMASTER, BARBARAG. SCHNEIDER, AND IZHAK NIR 42. Visualization of Intracellular Trafficking of Proteins RANDALE. MORRIS AND CATHARINE I . SAELINGER 43. Localization of Lectin Receptors RICHARD M. PINO 44. Localization of Gonadotropin-Releasing Hormone Receptors GWEN V. CHILDS 45. Localization of Ganglioside GMI with Biotinylated Choleragen HIROAKIASOU 46. Visualization of Adenosinetriphosphatase Site of Myosin Using Photoreactive Adenosine 5'-Diphosphate Analog KAZUO SUTOH

340 343 353 357 363 370 379 388 395 405 409

C. Protein Blotting 47. Analysis of Proteins and Glycoproteins on Blots EDWARD A. BAYER, HAYA BEN-HUR, AND MEIR WILCHEK 48. Direct Labeling of Blotted Glycoconjugates EDWARD A. BAYER, HAYA BEN-HUR, AND MEIR WILCHEK 49. Identification of Leukocyte Surface Proteins WALTER L. HURLEY AND EVE FINKELSTEIN 50. Staining of Proteins on Nitrocellulose Replicas WILLIAM J. LAROCHELLE AND STANLEY C. FROEHNER 51. Streptavidin-Enzyme Complexes in Detection of Antigens on Western Blots CHRISTINE L. BRAKEL, MARK S. BROWER, AND KIMBERLY GARRY

415 427 429 433 437

CONTENTS OF VOLUMES 168-174, 176-194 52. Luminescent Detection of Immunodot and Western Blots MERLIN M. L. LEONG AND GLYN R. FOX 53. Identification of Calmodulin-Binding Proteins MELVIN L. BILLINGSLEY, JOSEPH W. POLLI, KEITH R. PENNYPACKER, AND RANDALL L. KINCAID

xlvii 442

451

D. Immunoassays 54. Avidin-Biotin Mediated Immunoassays: Overview MEIR WILCHEK AND EDWARD A. BAYER 55. Avidin-Biotin System in Enzyme Immunoassays THI~R~:SE TERNYNCK AND STRATIS AVRAMEAS 56. Two-Site and Competitive Cbemiluminescent Immunoassays CHRISTIAN J. STRASBURGERAND FORTUNE KOHEN 57. Biotinylated Protein A in Immunoassay IKUO TAKASHIMA, NOBUO HASHIMOTO, AND HSHI-CHI CHANG 58. Slide lmmunoenzymatic Assay for Human Immunoglobulin E EVERLY CONWAY DE MACARIO, ROBERT J. JOVELL, AND ALBERTO J. L. MACARIO 59. Quantification of Carcinoembryonic Antigen in Serum and Analysis of Epitope Specificities of Monoclonal Antibodies CHRISTOPH WAGENER, LUCIA WICKERT, AND JOHN E. SHIVELY 60. Selective Precipitation of Biotin-Labeled Antigens or Monoclonal Antibodies by Avidin for Determining Epitope Specificities and Affinities in Solution-Phase Assays CHRISTOPHWAGENER, ULRICH KRUGER, AND JOHN E. SHIVELY 61. lmmunoassays for Diagnosis of Infectious Diseases ROBERT H. YOLKEN 62. Enzyme-Linked Immunosorbent Assay WILLIE F. VANN, ANN SUTTON, AND RACHEL SCHNEERSON 63. Rapid Detection of Herpes Simplex Virus LATA S. NERURKAR 64. Sensitive lmmunoassay for Human lmmunodeficiency Viral Core Proteins KENNETH R. HUSKINS, KEVIN J. REAGAN, JOHN A. WEHRLY, AND JEANNE E. NEUMANN

467 469 481 497

501

507

518 529 537 541

556

E. Gene Probes 65. Gene Probes MEIR WILCHEK AND EDWARD A. BAYER 66. Biotinylated Nucleotides for Labeling and Detecting DNA LEONARD KLEVAN AND GULILAT GEBEYEHU 67. Biotinylated Psoralen Derivative for Labeling Nucleic Acid Hybridization Probes COREY LEVENSON, ROBERT WATSON, AND EDWARD L. SHELDON 68. Chemically Cleavable Biotin-Labeled Nucleotide Analogs TIMOTHY M. HERMAN AND BARBARAJ. FENN 69. Preparation and Uses of Photobiotin JAMES L. MCINNES, ANTHONY C. FORSTER, DEREK C. SK1NGLE, AND ROBERT H. SYMONS 70. Modification of Cytosine Residues on DNA RAPHAEL P. VISCIDI 71. Direct Incorporation of Biotin into DNA MEIR WILCHEK, JEFFREY U. ROTHENBERG, Av1 REISFELD, AND EDWARD A. BAYER 72. Colorimetric-Detected DNA Sequencing STEPHAN BECK

560 561 577 584

588 600

608 612

xlviii

CONTENTS OF VOLUMES 168-174, 176-194

F. Composite and Special Applications 73, Composite Applications MEIR WILCHEK AND EDWARD A. BAYER 74. Third Component of Human Complement: C3 MELVIN BERGER 75. Purification and Characterization of Membrane Proteins JEAN-PIERRE KRAEHENBUHL AND CLAUDE BONNARD 76. Facilitated Cell Fusion for Hybridoma Production MARY K. CONRAD AND MATHEW M. S. Lo 77. Complex Neoglycoproteins MING-CHUAN SHAO, LING-MEI CHEN, AND FINN WOLD 78. Probing fl-Adrenergic Receptors KATHRYN E. MEIER AND ARNOLD E. RUOHO 79. Probing Adenosine Receptors Using Biotinylated Purine Derivatives KENNETH A. JACOaSON

618 619 629 641 653 660 668

VOLUME 185 G E N E EXPRESSION TECHNOLOGY

S e c t i o n I. I n t r o d u c t i o n I. Systems for Heterologous Gene Expression

DAVID V. GOEDDEL

S e c t i o n II. E x p r e s s i o n i n E s c h e r i c h i a coli 2. Expression of Heterologous Proteins in Escherichia coli

LARRY GOLD

II

3. Design and Construction of Expression Plasmid Vectors in Escherichia coli PAULINA BALBAS AND FRANCISCO BOLIVAR

14

A. Vectors

B. Promoters 4. Promoter Recognition and mRNA Initiation by Escherichia coli Eo"7° JAY O. GRALLA 5. Use of Escherichia coli trp Promoter for Direct Expression of Proteins DANIEL G. YANSURA AND DENNIS J. HENNER 6. Use of T7 RNA Polymerase to Direct Expression of Cloned Genes F. WILLIAM STUDIER, ALAN H. ROSENBERG, JOHN J. DUNN, AND JOHN W. DUBENDORFF

37 54

60

C. Translation 7. High-Level Translation Initiation LARRY GOLD AND GARY D. STORMO 8. Enhanced Translational Efficiency with Two-Cistron Expression System BRIGITTE E. SCHONER, RAMA M. BELAGAJE, AND RONALD G. SCHONER

89 94

CONTENTS OF VOLUMES 168-174, 1 7 6 - 1 9 4

9. Sequences within Ribosome Binding Site Affecting Messenger RNA Translatability and Method to Direct Ribosomes to Single Messenger RNA Species HERMAN A, DE BOER AND ANNA S. HuI 10. Vector for Enhanced Translation of Foreign Genes in Escherichia coli PETER O. OLINS AND SHAUKAT H. RANGWALA

xlix

103 115

D. Proteases 11. Minimizing Proteolysis in Escherichia coli: Genetic Solutions SUSAN GOTTESMAN

119

MATHIAS UHLl~N AND TOMAS MOKS

129 144 161

E. Gene Fusions 12. Gene Fusions for Purpose of Expression: An Introduction 13. Fusions to Staphylococcal Protein A 14. Expression as trpE Fusion

BJORN NILSSON AND LARS AaRAHMSl~N DANIEL G, YANSURA

F. Secretion 15. Engineering Escherichia coli to Secrete Heterologous Gene Products JOAN A. STADER AND THOMAS J. SILHAVY

166

G. Protein Refolding 16. Refolding of Recombinant Proteins TADAHIKO KOHNO, DAVID F. CARMICHAEL, ANDREAS SOMMER, AND ROBERT C. THOMPSON

187

S e c t i o n III. E x p r e s s i o n i n B a c i l l u s s u b t i l i s 17. Expression of Heterologous Genes in Bacillus subtilis

DENNISJ. HENNER

199

18. Regulated Promoter for High-Level Expression of Heterologous Genes in Bacillus subtilis STUART F. J. LE GR1CE

201

A. Intracellular

Production

B. Secretion 19. System for Secretion of Heterologous Proteins in Bacillus subtilis VASANTHA NAGARAJAN

214

C. Inducible Expression of Regulatory Proteins 20. Inducible Expression of Regulatory Genes in Bacillus subtilis DENNIS J. HENNER

223

1

CONTENTS OF VOLUMES

168-174, 176-194

Section IV. Expression in Yeast 21. Heterologous Gene Expression in Yeast

SCOTT D. E v a

231

A. Plasmid Vectors 22. Propagation and Expression of Cloned Genes in Yeast: 2-/zm Circle-Based Vectors ALAN B. ROSE AND JAMES R. BROACH 23. Manipulating Yeast Genome Using Plasmid Vectors TIM STEARNS, HONG MA, AND DAVID BOTSTE1N

234 280

B. Inducible Expression Cassettes 24. Regulated GAL4 Expression Cassette Providing Controllable and High-Level Output from High-Copy Galactose Promoters in Yeast LAWRENCE M. MYLIN, KATHRYN J. HOFMANN, LOREN D. SCHULTZ,AND JAMES E. HOPPER 25. Expression of Heterologous Proteins in Saccharomyces cereoisiae Using the ADH2 Promoter VIRGINIA L. PRICE, WAYNE E. TAYLOR, WILLIAM CLEVENGER, MARLIS WORTHINGTON, AND ELTON T. YOUNG 26. Induced Expression Using Yeast Copper Metallothionein Promoter TINA ETCHEVERRY

297

308 319

C. Constitutive Expression Cassettes 27. High-Efficiency Yeast Expression Vectors Based on the Promoter of the Phosphoglycerate Kinase Gene S. M. KINGSMAN, D. COUSENS, C. A. STANWAY, A. CHAMBERS, M. WILSON, AND A. J. KINGSMAN 28. Glyceraldehyde-3-phosphate Dehydrogenase-Derived Expression Cassettes for Constitutive Synthesis of Heterologous Proteins STEVEN ROSENBERG, DORIS COIT, AND PATRICIA TEKAMP-OLsoN 29. Superimposition of Temperature Regulation on Yeast Promoters ANDRZEJ Z. SLEDZIEWSKI, ANNE BELL, CARLI YIP, KIMBERLY KELSAY, FRANCIS J. GRANT, AND VIVIAN L. MACKAY

329

341

351

D. Translation 30. Sequence and Structural Requirements for Efficient Translation in Yeast THOMAS F. DONAHUE AND A. MARK CIGAN

366

E. Proteases 31. Vacuolar Proteases in Yeast Saccharomyces cerevisiae ELIZABETH W. JONES 32. Detection and Inhibition of Ubiquitin-Dependent Proteolysis KEITH D. WILKINSON 33. Cotranslational Amino-Terminal Processing RICHARD L. KENDALL, RYO YAMADA, AND RALPH A. BRADSHAW

372 387 398

CONTENTS OF VOLUMES 168-174, 176-194

li

F. Protein Secretion and Modification 34. a-Factor Leader-Directed Secretion of Heterologous Proteins from Yeast ANTHONY J. BRAKE 35. Use of Heterologous and Homologous Signal Sequences for Secretion of Heterologous Proteins from Yeast RONALD A. HITZEMAN, CHRISTINA Y. CHEN, DONALD J. DOWBENKO, MARK E. RENZ, CHUNG LIU, ROGER PAl, NANCY J. SIMPSON, WILLIAM J. KOHR, AR.IUN SINGH, VANESSA CHISHOLM, ROBERT HAMILTON, AND CHUNG NAN CHANG 36. Isolation, Characterization, and Properties of S a c c h a r o m y c e s cerevisiae rnnn Mutants with Nonconditional Protein Glycosylation Defects CLINTON E. BALLOU 37. Molecular and Genetic Approach to Enhancing Protein Secretion VANESSA CHISHOLM, CHRISTINA Y. CHEN, NANCY J. SIMPSON, AND RONALD A. HITZEMAN

S e c t i o n V. E x p r e s s i o n i n M a m m a l i a n

408

421

440

471

Cells

38. Expression of Heterologous Genes in Mammalian Cells ARTHUR D. LEVINSON

485

39. Vectors Used for Expression in Mammalian Cells RANDALJ. KAUFMAN 40, Assembly of Enhancers, Promoters, and Splice Signals to Control Expression of Transferred Genes MICHAEL KRIEGLER

487

A. Vectors

512

B. Transfection Methods 41. Methods for Introducing DNA into Mammalian Cells WAYNE m. KEOWN, COLIN R. CAMPBELL, AND RAJU S. KUCHERLAPATI

527

C. Markers for Selection and Amplification 42. Selection and Coamplification of Heterologous Genes in Mammalian Cells RANDAL J. KAUFMAN

537

D. Growth of Cell Lines 43. Optimizing Cell and Culture Environment for Production of Recombinant Proteins JENNIE P. MATHER

567

E. Posttranslation Processing, Modification, and Secretion 44. Analysis of Synthesis, Processing, and Secretion of Proteins Expressed in Mammalian Cells ANDREW J. DORNER AND RANDAL J. KAUFMAN

577

lii

CONTENTS OF VOLUMES 168-174, 176-194

Section VI. M u t a g e n e s i s 45. Saturation Mutagenesis Using Mixed Oligonucleotides and M13 Templates Containing Uracil BRUCE H. HORWITZAND DANIELDIMAIO

599

V O L U M E 186 OXYGEN RADICALS IN BIOLOGICAL SYSTEMS PART B OXYGEN RADICALS AND ANTIOXIDANTS

1. Role of Free Radicals and Catalytic Metal Ions in Human Disease: An Overview BARRY HALLIWELL AND JOHN M. C. GUTTERIDGE

S e c t i o n I. P r o d u c t i o n , D e t e c t i o n , a n d C h a r a c t e r i z a t i o n of O x y g e n R a d i c a l s a n d O t h e r Species 2. Pulse Radiolysis in Study of Oxygen Radicals MICHAELG. S|MIC 3. Free Radical Initiators as Source of Water- or Lipid-Soluble Peroxyl Radicals

89

ETSUO NIKI

100

4. Generation of Iron(IV) and Iron(V) Complexes in Aqueous Solutions BENON H. J. BIELSKi

108

5. Sources of Compilations of Rate Constants for Oxygen Radicals in Solution ALBERTA B. Ross AND BENON H. J. BIELSKI 6. Use of Polyaminocarboxylates as Metal Chelators

113

DIANE E. CABELLI AND BENON H. J. BIELSKI

116

7. Preparation of Metal-Free Solutions for Studies of Active Oxygen Species K. M. SCHMCH 8. Use of Ascorbate as Test for Catalytic Metals in Simple Buffers

121

GARRY R. BUETTNER

125

9. Spin-Trapping Methods for Detecting Superoxide and Hydroxyl Free Radicals in Vitro and in Vivo GARRY R. BUETTNER AND RONALD P. MASON 10. Detection of Singlet Molecular Oxygen during Chloride Peroxidase-Catalyzed Decomposition of Ethyl Hydroperoxide ROBERT D. HALL 11. Detection and Quantitation of Hydroxyl Radical Using Dimethyl Sulfoxide as Molecular Probe

CHARLES F. BABBS AND MELISSA GALE STEINER

127

133 137

12. Distinction between Hydroxyl Radical and Ferryl Species J. D. RUSH, g. MASKOS, AND W. H. KOPPENOL

148

13. Determination of Hydroperoxides with Fluorometric Reagent Diphenyl-lpyrenylphosphine HIROSHI MEGURO, KAZUAKI AKASAKA,AND HIROSHI OHRUI

157

14. Phycoerythrin Fluoroscence-Based Assay for Reactive Oxygen Species ALEXANDER N. GLAZER

161

15. Sulfur-Centered Free Radicals KLAUS-DIETERASMUS 16. Quinoid Compounds: High-Performance Liquid Chromatography with Electrochemical Detection ENRIQUE CADENAS AND LARS ERNSTER

168 180

CONTENTS OF VOLUMES 168-174, 176-194 17. Preparation of Tocopheroxyl Radicals for Detection by Electron Spin Resonance ROEF J. MEHLHORN, JORGEN FUCHS, SATOSHI SUMIDA, AND LESTER PACKER

S e c t i o n II. A s s a y o f E n z y m e s o r S u b s t a n c e s I n v o l v e d i n F o r m a t i o n Removal of Oxygen Radicals and Derived Products

liii

197

or

A. Proteins 18. Determination of Superoxide Dismutase Activity by Purely Chemical System Based on NAD(P)H Oxidation FRANCESCO PAOLETTI AND ALESSANDRA MOCAL! 19. Assays for Superoxide Dismutase Based on Autoxidation of Hematoxylin JOSEPH P. MARTIN, JR. 20. Assay for Superoxide Dismutase Based on Chemiluminescence of Luciferin Analog MINORU NAKANO 21. Automated Assay of Superoxide Dismutase in Blood MARY R. L'ABBf~ AND PETER W. F. FISCHER 22. Assay of Superoxide Dismutase Applicable to Whole Bacterial Cells F. S. ARCHIBALD 23. Superoxide Dismutase Mimic Prepared from Desferrioxamine and Manganese Dioxide WAYNE F. BEYER, JR. AND IRWIN FRIDOVICH 24. Purification of Exocellular Superoxide Dismutases KENNETHD. MUNKRES 25. Analysis of Extracellular Superoxide Dismutase in Tissue Homogenates and Extracellular Fluids STEFAN L. MARKLUND 26. Oxidative Reactions of Hemoglobin CHRISTINE C. WINTERBOURN 27. Assays for Cytochrome P-450 Peroxygenase Activity MINOR J. COON, ROBERT C. BLAKE I1, RONALD E. WHITE, AND GERALD D. NORDBLOM 28. Reductive Cleavage of Hydroperoxides by Cytochrome P-450 AEFIN D. N. VAZ AND MINOR J. COON 29. Prostaglandin H Synthase JEFF A. BOYD 30. DT-Diaphorase: Purification, Properties, and Function CHRISTINA LIND, ENRIQUE CADENAS, PAUL HOCHSTEIN, AND LARS ERNSTER

209 220 227 232 237 242 249 260 265

273 278 283 287

B. Small Molecules 31. Antioxidant Activities of Bile Pigments: Biliverdin and Bilirubin ROLAND STOCKER, ANTONY F. McDONAGH, ALEXANDER N. GLAZER, AND BRUCE N. AMES 32. Ergothioneine as Antioxidant PHILIP E. HARTMAN 33. Thiyl Free Radical Metabolites of Thiol Drugs, Glutathione, and Proteins RONALD P. MASON AND D. N. RAMAKRISHNA RAO 34. Estrogens as Antioxidants ETsuo NIKI AND MINORU NAKANO 35. Characterization of o-Semiquinone Radicals in Biological Systems B. KALYANARAMAN 36. Flavonoids as Antioxidants: Determination of Radical-Scavenging Efficiencies WOLF BORS, WERNER HEELER, CHRISTA MICHEL, AND MANFRED SARAN

301 310 318 330 333 343

liv

CONTENTS OF VOLUMES 168-174, 176-194

37. Fluorescence Measurements of Incorporation and Hydrolysis of Tocopherol and Tocopheryl Esters in Biomembranes VALERIAN E. KAGAN, RUMYANA A. BAKALOVA, ELENA E. SERBINOVA, AND TSANKO S. STOYTCHEV

355

Section III. Assay and Repair of Biological Damage Due to Oxygen and Oxygen-Derived Species A. Lipid Peroxidation 38. Assay of Lipid Hydroperoxides Using High-Performance Liquid Chromatography with Isoluminal Chemiluminescence Detection YORIHIRO YAMAMOTO, BALZ FREI, AND BRUCE N. AMES 39. Determination of Methyl Linoleate Hydroperoxides by t3C Nuclear Magnetic Resonance Spectroscopy E. N. FRANKEL, W. E. NEFF, AND D. WEISLEDER 40. Gas Chromatography-Mass Spectrometry Assays for Lipid Peroxides FREDERIK J. G. M. VAN KUIJK, DAVID W. THOMAS, ROaERT J. STEPHENS, AND EDWARD A. DRATZ 41. Gas Chromatography-Mass Spectrometry of 4-Hydroxynonenal in Tissues FREDERIK J. G. M. VAN KUIJK, DAVID W. THOMAS, ROBERT J. STEPHENS, AND EDWARD A. DRATZ 42. Determination of Aldehydic Lipid Peroxidation Products: Malonaldehyde and 4Hydroxynonenal HERMANNESTERBAUER AND KEVIN H. CHEESEMAN 43. Malondialdehyde Determination as Index of Lipid Peroxidation H. H. DRAPER AND M. HADLEY 44. Cyclooxygenase Initiation Assay for Hydroperoxides RICHARD J. KULMACZ, JAMES F. MILLER, JR., ROBERT B. PENDLETON, AND WILLIAM E. M. LANDS 45. Cholesterol Epoxides: Formation and Measurement G. A. S. ANSARI AND LELAND L. SMITH 46. Oxystat Technique in Study of Reactive Oxygen Species HERBERT DE GROOT 47. Phospholipid Hydroperoxide Glutathione Peroxidase MATILDE MAIORINO, CARLO GREGOLIN, AND FULVIO URSINI 48. Iron Redox Reactions and Lipid Peroxidation STEVEN D. AUST, DENNIS M. MILLER, AND VICTOR M. SAMOKYSZYN

371

380

388

399 407 421

431 438 443 448 457

B. Protein Oxidation 49. Determination of Carbonyl Content in Oxidatively Modified Proteins RODNEY L. LEVINE, DONITA GARLAND, CYNTHIA N. OLIVER, ADOLFO AMICI, ISABEL CLIMENT, ANKE-G. LENZ, BONG-WHAN AHN, SHMUEL SHALTIEL, AND EARL R. STADTMAN 50. Protein That Prevents Mercaptan-Mediated Protein Oxidation SUE GOD RHEE, KANGHWA KIM, IL HAN KIM, AND EARL R. STADTMAN 51. Protein Degradation as an Index of Oxidative Stress ROBERT E. PACIFICI AND KELVIN J. A. DAVIES

464 478 485

CONTENTS OF VOLUMES 168-174, 176-194

Iv

C. Modification of Nucleic Acids and Chromosomes 52. Photodynamic Methods for Oxy Radical-Induced DNA Damage J. R. WAGNER, J. E. VAN LIER, C. DECARROZ, M. BERGER, AND J. CADET 53. Radical-Mediated DNA Damage in Presence of Oxygen

502

CLEMENS VON SONNTAG AND HEINZ-PETER SCHUCHMANN

511

54. In VivoOxidativeDNADamage: Measurement of 8-Hydroxy-2'-deoxyguanosine in DNA and Urine by High-Performance Liquid Chromatography with Electrochemical Detection MARK SHIGENAGA, JEEN-Woo PARK, KENNETH C. CURDY, CARLOS J. GIMENO, AND BRUCE N. AMES

521

55. Selected-Ion Mass Spectrometry: Assays of Oxidative DNA Damage MIRAL DIZDAROGLU AND EWA GAJEWSKI

530

56. Footprinting Protein-DNA Complexes with y-Rays JEFFREY J. HAYES, LAURANCE KAM, AND THOMAS D. TULLIUS

545

57. Fluorometric Analysis of DNA Unwinding to Study Strand Breaks and Repair in Mammalian Cells H. CHAIM BIRNBOIM 58. Clastogenic Factors: Detection and Assay INGRID EMER1T

550 555

S e c t i o n IV. L e u k o c y t e s a n d M a c r o p h a g e s 59. Kinetic Micro#ate Assay for Superoxide Production by Neutrophils and Other LAURA A. MAYO AND JOHN T. CURNUTTE

567

60. Measurement of Oxidized Proteins in Systems Involving Activated Neutrophils or HL-60 Cells CYNTHIA N. OLIVER 61. Xenobiotic Activation by Stimulated Human Polymorphonuclear Leukocytes

Phagocytic Cells

575

and Myeloperoxidase

DAVID A. EASTMOND AND MARTYR T. SMITH

62. Determination of Superoxide Radical and Singlet Oxygen Based on Chemiluminescence of Luciferin Analogs MINORU NAKANO

579

585

S e c t i o n V. O r g a n , T i s s u e , a n d Cell D a m a g e a n d M e d i c a l A p p l i c a t i o n s 63. Visible-Range Low-Level Chemiluminescence in Biological Systems MICHAEL n . MURPHY AND HELMUT SIES

64. Spin Trapping Biologically Generated Free Radicals: Correlating Formation with Cellular Injury GERALD M. ROSEN AND HOWARDJ. HALPERN 65. Oxygen Radicals and Drugs: In Vitro Measurements D. M. ZIEGLER AND JAMES P. KEHRER 66. Modulation of Cellular Glutathione and Protein Thiol Status during Quinone Metabolism

GIORGIO BELLOMO, HJORDIS THOR, AND STEN ORRENIUS

595

611 621 627

67. Production of Oxygen Radicals by Photosensitization JOSEPH P. MARTIN, JR. AND PAULA BURCH

635

68. Elevated Mutagenesis in Bacterial Mutants Lacking Superoxide Dismutase DANIELE TOUATI AND SPENCER B. FARR

646

69. Measurement of Xanthine Oxidase in Biological Tissues LANCE S. TERADA, JONATHAN m. LEFF, AND JOHN E. REPINE

651

70. Oxy Radicals in Disseminated Intravascular Coagulation TOSHIKAZU YOSHIKAWA

656

lvi

CONTENTS OF VOLUMES 168-174, 176-194

71. Oxy Radicals in Endotoxin Shock TOSHIKAZU YOSHIKAWA 72. Preparation of Human Lung Tissue from Cigarette Smokers for Analysis by Electron Spin Resonance Spectroscopy DANIEL F. CHURCH, THEODORE J. BURKEY, AND WILLIAM A. PRYOR 73. Assay for Free Radical Reductase Activity in Biological Tissue by Electron Spin Resonance Spectroscopy JURGEN FUCHS, ROLE J. MEHLHORN, AND LESTER PACKER 74. Biochemical Pharmacology of Inflammatory Liver Injury in Mice ALBRECHT WENDEL 75. Measurement of Oxidant Stress in Vivo HELEN HUGHES, HARTMUT JAESCHKE, AND JERRY R. MITCHELL 76. Radioprotection by Ascorbic Acid, Desferal, and Mercaptoethylamine AJIT SINGH, HARWANT SINGH, AND JAMES S. HENDERSON 77. Influence of Thiols on Thermosensitivity of Mammalian Cells in Vitro ROLF D. ISSELS AND ARNO NAGELE

660

665

670 675 681 686 696

Section VI. Oxygen Free Radicals in Ischemia and

Reperfusion-Associated Tissue Damage 78. Oxygen-Derived Free Radicals in Repeffusion Injury MARC S. SUSSMAN AND GREGORY B. BULKLEY 79. Noninvasive Fluorometric Measurement of Mitochondrial Membrane Potential in Isolated Working Rat Hearts during Ischemia and Repeffusion JORGEN FUCHS, G. ZIMMER, T. THORICH, J. BERE1TER-HAHN, AND LESTER PACKER 80. Assessment of Leukocyte Involvement during Ischemia and Reperfusion of Intestine MATTHEW B. GRISHAM, JOSEPH N. BENOIT, AND D. NElL GRANGER 81. Oxygen Scavengers in Myocardial Preservation during Transplantation JAMES R. STEWART, WILLIAM H. FRIST, AND WALTER H. MERRILL 82. Quantitation of Free Radical-Mediated Reperfusion Injury in Renal Transplantation G. MELVILLE WILLIAMS 83. Use of Isolated Perfused Organs in Hypoxia and Ischemia/Reperfusion Oxidant Stress HARTMUT JAESCHKE AND JERRY R. MITCHELL 84. Use of Perfused Organs in Measurement of Drug-lnduced Oxidant Stress RAYMOND F. BURK AND KRIST1NA E. HlLL

VOLUME 187 A R A C H I D O N A T E R E L A T E D L I P I D MEDIATORS 1. Nomenclature WILLIAM L. SMITH, PIERRE BORGEAT, MATS HAMBERG, L. JACKSON ROBERTS II, ANTHONY L. WILLIS, SHOZO YAMAMOTO, PETER W. RAMWELL, JOSHUA ROKACH, BENGT SAMUELSSON, E. J. COREY, AND C. R. PACE-ASCIAK

711

723

729 742 748 752 759

CONTENTS OF VOLUMES 168-174, 176-194

Ivii

Section I. Assays

A. P r o s t a g l a n d i n s 2. Electron-Capture Negative-Ion Chemical Ionization Mass Spectrometry of Lipid Mediators IAN A. BLAIR 3. Enzyme Immunoassays of Eicosanoids Using Acetylcholinesterase

13

PHILIPPE PRADELLES, JACQUES GRASSI, AND JACQUES MACLOUF

24

4. Radioimmunoassay of l l-Dehydrothromboxane B2 GIOVANNI CIABATTONI, PAOLA PATRIGNANI, AND CARLO PATRONO

5. Measurement of Thromboxane Metabolites by Gas Chromatography-Mass Spectrometry FRANCESCA CATELLA AND GARRET A. FITZGERALD 6. Quantification of 9ct,1 lfl-Prostaglandin F2 by Stable Isotope Dilution Mass Spectrometric Assay DANIEL F. WENDELBORN, JASON D. MORROW, AND L. JACKSON ROBERTS I1

34 42

51

7. Immunoaffinity Purification-Chromatographic Quantitative Analysis of Arachidonic Acid Metabolites J. JAMES VRBANAC, JEFFREY W. Cox, THOMAS D. ELLER, AND DANIEL R. KNAPP

62

B. L e u k o t r i e n e s 8. Preparation of Tetradeuterated Leukotriene A4 Methyl Ester: Methyl[I I, 12,14,15-2H4]-(5S,6S)-Oxido-(7E,9E,IIZ, 14Z)-Eicosatetraenoate J. P. LELLOUCHE,J. P. BEAUCOURT,AND A. VANHOVE 9. Quantitative Gas Chromatography-Mass Spectrometry Analysis of Leukotriene B4 W. RODNEY MATHEWS 10. Enzyme Immunoassays for Leukotrienes C4 and E4 Using Acetylcholinesterase PHILIPPE PRADELLES, CATHERINEANTOINE, JEAN-PAULLELLOUCHE, AND JACQUES MACLOUF

11. Quantitation of Sulfidopeptide Leukotrienes in Biological Fluids by Gas Chromatography-Mass Spectrometry R O B E R TC. MURPHY AND ANGELO SALA 12. Automated On-Line Extraction and Profiling of Lipoxygenase Products of Arachidonic Acid by High-Performance Liquid Chromatography PIERRE BORGEAT, SERGE PICARD, PIERRE VALLERAND, SYLVAIN BOURGOIN. ABDULRAHMAN ODEIMAT, PIERRE SIROIS, AND PATRICE E. POUBELLE

13. Preparation of Antibodies Directed against Leukotrienes

70 76

82 90

98

EDWARDC. HAYES

116

MARIE-JEANNE BOSSANT, EWA NIN10, DANII~LE DELAUTIER, AND JACQUES BENVENISTE

125

C. P l a t e l e t - A c t i v a t i n g F a c t o r 14. Bioassay of paf-Acether by Rabbit Platelet Aggregation

15. Bioassay of Platelet-Activating Factor by Release of [3H]-Serotonin PETER M. HENSON

16. Quantitation of Platelet-Activating Factor by Gas Chromatography-Mass Spectrometry KEITH L. CLAY 17. Quantitative Analysis of Platelet-Activating Factor by Gas ChromatographyNegative-lon Chemical Ionization Mass Spectrometry WALTER C. PICKETT AND CHAKKODABYLUS. RAMESHA

130

134

142

lviii

CONTENTS OF VOLUMES 168-174, 176-194

18. Isolation of Platelet-Activating Factor and Purification by Thin-Layer Chromatography DONALD J. HANAHAN 19. Separation and Characterization of Arachidonate-Containing Phosphoglycerides FLOYD H. CHILTON

152 157

D. Other Oxidative Products 20. High-Performance Liquid Chromatography Separation and Determination of Lipoxins CHARLES N. SERHAN 21. Quantitation of Epoxy- and Dihydroxyeicosatrienoic Acids by Stable IsotopeDilution Mass Spectrometry JOHN TURK, W. THOMAS STUMP, WENDY CONRAD-KESSEL, ROBERT R. SEABOLD, AND BRYAN A. WOLF 22. High-Performance Liquid Chromatography for Chiral Analysis of Eicosanoids ALAN R. BRASH AND DAN J. HAWKINS

167

175 187

E. Phospholipase A2 and Phospholipids 23. Extraction of Phospholipids and Analysis of Phospholipid Molecular Species GEORGE M. PATTON AND SANDERJ. ROBINS 24. Macrophage Phospholipase A2 Specific for sn-2-Arachidonic Acid CHRISTINA C. LESLIE 25. Measurement of Phosphoinositide-Specific Phospholipase C Activity JOHN E. BLEASDALE,JAMES C. McGUIRE, AND GREGORYA. BALA 26. Solubilization of Arachidonate-CoA Ligase from Cell Membranes, Chromatographic Separation from Nonspecific Long-Chain Fatty Acid CoA Ligase, and Isolation of Mutant Cell Line Defective in Arachidonate-CoA Ligase MICHAEL LAPOSATA

195 216 226

237

Section II. Biosynthesis, Enzymology, and Chemical Synthesis A. Prostaglandins 27. Preparation of Prostaglandin H2: Extended Purification/Analysis Scheme DUANE VENTON, GuY LE BRETON, AND ELIZABETHHALL 28. Purification and Properties of Pregnancy-Inducible Rabbit Lung Cytochrome P-450 Prostaglandin to-Hydroxylase A. SCOTT MUERHOFF, DAVID E. WILLIAMS, AND BETTIE SUE SILER MASTERS

245

253

B. Leukotrienes 29. Purification of Arachidonate 5-Lipoxygenase from Potato Tubers PALLU REDDANNA, J. WHELAN, K. R. MADDIPATI, AND C. CHANNA REOOY 30. Leukotriene Metabolism by Isolated Rat Hepatocytes MICHAEL A. SHIRLEY AND DANNY O. STENE 31. Purification and Characterization of Human Lung Leukotriene A4 Hydrolase NOBUYA OHISHI, TAKASHI IZUMI, YOUSUKE SEYAMA, AND TAKAO SHIMIZU

268 277 286

CONTENTS OF VOLUMES

168-174, 176-194

32. Potato Arachidonate 5-Lipoxygenase: Purification, Characterization, and Preparation of 5(S)-Hydroperoxyeicosatetraenoic Acid TAKAO SHIMIZU, ZEN-ICHIRO HONDA, ICHIRO MIKI, YOUSUKE SEYAMA, TAKASHI IZUMI, OLOF R~,DMARK, AND BENGT SAMUELSSON 33. Leukotriene Co Synthase: Characterization in Mouse Mastocytoma Cells MATS SODERSTROM,BENGT MANNERVlK, AND SVEN HAMMARSTROM 34. Leukocyte Arachidonate 5-Lipoxygenase: Isolation and Characterization CAROL A. ROUZER AND BENGT SAMUELSSON 35. Cytochrome P-450LTB and Inactivation of Leukotriene B4 RoY J. SOBERMAN 36. Cytosolic Liver Enzymes Catalyzing Hydrolysis of Leukotriene A4 to Leukotriene B4 and 5,6-Dihydroxyeicosatetraenoic Acid JESPERZ. HAEGGSTROM 37. Purification and Properties of Leukotriene C4 Synthase from Guinea Pig Lung Microsomes RoY J. SOBERMAN 38. Immunoaffinity Purification of Arachidonate 5-Lipoxygenase from Porcine Leukocytes NATSUO UEDA AND SHOZO YAMAMOTO

lix

296 306 312 319 324 335 338

C. Platelet-Activating Factor 39. Platelet-Activating Factor Acetylhydrolase from Human Plasma DIANA M. STAFFORINI, THOMAS M. MCINTYRE, AND STEPHEN M. PRESCOTT

344

D. Other Oxidative Products 40. Synthesis of Epoxyeicosatrienoic Acids and Heteroatom Analogs J. R. FALCK, PENDRI YADAGIRI, AND JORGE CAPDEVILA 41. Isolation of Rabbit Renomedullary Cells and Arachidonate Metabolism MAIREAD A. CARROLL, ELIZABETH D. DRUGGE. CATHERINE E. DUNN, AND JOHN C. MCGIFF 42. Ocular Cytochrome P-450 Metabolism of Arachidonate: Synthesis and Bioassay MICHAL LANIADO SCHWARTZMAN AND NADER G. ABRAHAM 43. Cytochrome P-450 Arachidonate Oxygenase JORGE H. CAPDEVILA, J. R. FALCK, ELIZABETH DISHMAN, AND ARMANDO KARARA

357

365 372

385

Section III. Pharmacology: Antagonist and Synthesis Inhibitors A. Prostaglandins 44. Radioligand Binding Assays for Thromboxane ACProstaglandin H2 Receptors PERRY V. HALUSHKA, THOMAS A. MORINELLI, AND DALE E. MAIS 45. Thromboxane A2/Prostaglandin Hz Receptor Antagonists GuY C. LE BRETON, CHANG T. LIM, CHITRA M. VAIDYA, AND DUANE L. VENTON

397

406

B. Leukotrienes 46. Assessment of Leukotriene D4 Receptor Antagonists 47. Sulfidopeptide Leukotriene Receptor Binding Assays

DAVID AHARONY SEYMOUR MONG

414 421

Ix

CONTENTS OF VOLUMES 168-174, 176-194

C. Platelet-Activating Factor 48. Ginkgolides and Platelet-Activating Factor Binding Sites D. J. HOSFORD, M. T. DOMINGO, P. E. CHABRIER,AND P. BRAQUET 49. Kadsurenone and Other Related Lignans as Antagonists of Platelet-Activating Factor Receptor T . Y . SHEN AND I. M. HUSSAINI 50. Use of WEB 2086 and WEB 2170 as Platelet-Activating Factor Antagonists JORGE CASALS-STENZEL AND HUBERT O. HEUER

433 446 455

Section IV. Molecular Biology A. Prostaglandins 51. Cloning of Sheep and Mouse Prostaglandin Endoperoxide Synthases DAVID L. DEWITT AND WILLIAM L. SMITH 52. Preparation and Proteolytic Cleavage of Apoprostaglandin Endoperoxide Synthase REBECCA ODENWALLER, YING-NAN PAN CHEN, AND LAWRENCE J. MARNETT

469

479

B. Leukotrienes 53. Cloning of Leukotriene A4 Hydrolase cDNA OLOF R/~DMARK, COLIN FUNK, Jl YI Fu, TAKASHI MATSUMOTO, HANS JORNVALL, BENGT SAMUELSSON, M1CHIKO M1NAMI, SHIGEO OHNO, HIROSHI KAWASAKI, YOUSUKE SEYAMA, KOICHI SUZUKI, AND TAKAO SH1MIZU 54. Molecular Biology and Cloning of Arachidonate 5-Lipoxygenase COLIN D. FUNK, TAKASHI MATSUMOTO, AND BENGT SAMUELSSON

486 491

Section V. Cell Models of Lipid Mediator Production A. Isolated Cell Preparations 55. Parietal Cell Preparation and Arachidonate Metabolism N. ANN PAYNE AND JOHN G. GERBER 56. Culture of Bone Marrow-Derived Mast Cells: A Model for Studying Oxidative Metabolism of Arachidonic Acid and Synthesis of Other Molecules Derived from Membrane Phospholipids EHUD RAZIN 57. Endothelial Cells for Studies of Platelet-Activating Factor and Arachidonate Metabolites G u y A. ZIMMERMAN,RALPH E. WHATLEY,THOMAS M. MCINTYRE, DONELLE i . BENSON, AND STEPHEN M. PRESCOTT 58. Use of Cultured Cells to Study Arachidonic Acid Metabolism ROBERT R. GORMAN, MICHAEL J. BIENKOWSKI, AND CHRISTOPHER W. BENJAMIN 59. Eicosanoid Biochemistry in Cultured Glomerular Mesangial Cells MICHAEL S. SIMONSON AND MICHAEL J. DUNN

505

514

520

535 544

CONTENTS OF VOLUMES 168--174, 176--194

lxi

B. Cell-Cell Interactions 60. Leukotriene B4 Biosynthesis by Erythrocyte-Neutrophil Interactions DAVID A. JONES AND FRANK A. FlrZPATmCK 61. Leukotriene C4 Biosynthesis during Polymorphonuclear Leukocyte-Vascular Cell Interactions STEVEN J. FEINMARK 62. Release and Metabolism of Leukotriene A4 in Neutrophil-Mast Cell Interactions CLEMENS A. DAHINDEN AND URS WIRTHMUELLER 63. Interaction between Platelets and Lymphocytes in Biosynthesis of Prostacyclin KENNETH K. Wu AND AUDREY C. PAPP 64. Eicosanoid Interactions between Platelets, Endothelial Cells, and Neutrophils AARON J. MARCUS

553 559 567 578 585

C. In Vitro Tissue Preparations 65. Isolated Perfused Rat Lung in Arachidonate Studies SHIH-WEN CHANG AND NORBERT F. VOELKEL 66. Isolated Coronary-Peffused Mammalian Heart: Assessment of Eicosanoid and Platelet-Activating Factor Release and Effects ROBERTO LEVI AND KEV1N M. MULLANE 67. Preparation of Human and Animal Lung Tissue for Eicosanoid Research T. VIGAN(~, M. T. CR1VELLARI,M. MEZZETTI, AND G. C. FOLCO

599

610 621

VOLUME 188 HYDROCARBONS AND METHYLOTROPHY

Section I. Hydrocarbons and Related Compounds A. Enzymes of Alkane and Alkene Utilization 1. Hydrocarbon Monooxygenase System of Pseudomonas oleovorans SHELDON W. MAY AND ANDREAS G. KATOPODIS 2. Assay Methods for Long-Chain Alkane Oxidation in Aeinelobaeter W. R. FINNERTY 3. Primary Alcohol Dehydrogenases from Acinetobacter W . R . FINNERTY 4. Aldehyde Dehydrogenases from Acinetobacter W . R . FINNERTY 5. Propane-Specific Alcohol Dehydrogenase from Rhodococcus rhodochrous PNKbl W. ASHRAF AND J. C. MURRELL 6. Cell-Free Assay Methods for Enzymes of Propane Utilization J. C. MURRELL AND W. ASHRAF 7. Quinoprotein Alcohol Dehydrogenase from Pseudomonas aeruginosa and Quinohemoprotein Alcohol Dehydrogenase from Pseudornonas testosteroni B. W. GROEN AND J. A. DUINE

3 10 14 18 21 26

33

lxii

CONTENTS OF VOLUMES 168-174, 176-194

B. Enzymes Involved in Cyclic and Aromatic Hydrocarbon Utilization 8. Toluene Dioxygenase from Pseudomonas putida FI LAWRENCE P. WACKETT 9. Naphthalene Digxygenase from Pseudomonas NCIB 9816 BURT D. ENSLEY AND BILLY E. HAIGLER 10. Benzene Dioxygenase from Pseudomonas putida ML2 (NCIB 12190) PHILIP J. GEARY, JEREMY R. MASON, AND CHRIS L. JOANNOU 11. Phthalate Dioxygenase CHRISTOPHER J. BATIE AND DAVID P. BALLOU 12. Cyclohexanone 1,2-Monooxygenase from Acinetobacter NCIMB 9871 PETER W. TRUDGILL 13. Cyclopentanone 1,2-Monooxygenase from Pseudomonas NCIMB 9872 PETER W. TRUDGILL 14. Protocatechuate 3,4-Dioxygenase from Brevibacterium fuscum JAMES W. WHITTAKER, ALLEN M. ORVILLE, AND JOHN D. LIPSCOMB 15. Protocatechuate 4,5-Dioxygenase from Pseudomonas testosteroni DAVID M. ARCIERO, ALLEN M. ORVILLE, AND JOHN D. LIPSCOMB 16. Protocatechuate 2,3-Dioxygenase from Bacillus macerans SANFORD A. WOLGEL AND JOHN D. LIPSCOMB 17. Gentisate 1,2-Dioxygenase from Pseudomonas acidovorans MARK R. HARPEL AND JOHN D. LIPSCOMB 18. Synthesis of ~70- or ~80-Enriched Dihydroxy Aromatic Compounds ALLEN M. ORVILLE, MARK R. HARPEL, AND JOHN D. LIPSCOMB 19. Catechol 2,3-Dioxygenases from Pseudomonas aeruginosa 2x I. A. KATAEVA AND L. g. GOLOVLEVA 20. Catechol and Chlorocatechol 1,2-Dioxygenases KA-LEUNG NGAI, ELLEN L. NEIDLE, AND L. NICHOLAS ORNSTON 21. Muconate Cycloisomerase RICHARD B. MEAGHER, KA-LEUNG NGAI, AND L. NICHOLAS ORNSTON 22. Muconolactone Isomerase RICHARD B. MEAGHER, KA-LEUNG NGAI, AND L. NICHOLAS ORNSTON 23. cis-l,2-Dihydroxycyclohexa-3,5-diene (NAD) Oxidoreductase (cis-Benzene Dihydrodiol Dehydrogenase) from Pseudornonas putida NCIB 12190 JEREMY R. MASON AND PHILIP J. GEARY 24. Hydroxybenzoate Hydroxylase BARRIE ENTSCH 25. Polycyclic Aromatic Hydrocarbon Degradation by Mycobacterium CARL E. CERNIGLIA AND MICHAEL A. HEITKAMP 26. Benzoate-CoA Ligase from Rhodopseudomonas palustris JANE GIBSON, JOHANNA F. GEISSLER, AND CAROLINE S. HARWOOD

39 46 52 61 70 " 77 82 89 95 101 107 115 122 126 130

134 138 148 154

C. Yeast and Fungal Enzymes of Hydrocarbon Degradation 27. Lignin Peroxidase from Fungi: Phanerochaete chrysosporium T. KENT KIRK, MING TIEN, PHILIP J. KERSTEN, B. KALYANARAMAN, KENNETH E. HAMMEL, AND ROBERTA L. FARRELL 28. Long-Chain Alcohol Dehydrogenase of Candida Yeast MITSUYOSHI UEDA AND ATSUO TANAKA 29. Long-Chain Aldehyde Dehydrogenase of Candida Yeast MITSUYOSHi UEDA AND ATSUO TANAKA

159 171 176

CONTENTS OF VOLUMES 168-174, 1 7 6 - 1 9 4

lxiii

S e c t i o n II. M e t h y l o t r o p h y

A. D i s s i m i l a t o r y E n z y m e s o f M e t h y l o t r o p h i c B a c t e r i a 30. Soluble Methane Monooxygenase from Methylococcus capsulatus Bath SIMON J. PILKINGTON AND HOWARD DALTON

181

31. Methane Monooxygenase from Methylosinus trichosporium OB3b BRIAN G. F o x , WAYNE A. FROLAND, DAVID R. JOLLIE, AND JOHN D. LIPSCOMB

191

32. Methanol Dehydrogenase from Hyphomicrobium X J. FRANKAND J. A. DUINE 33. Methanol Dehydrogenase from Methylobacterium extorquens AMI

202

DARREN J. DAY AND CHRISTOPHER ANTHONY

210

34. Modifier Protein for Methanol Dehydrogenase of Methylotrophs ANTONY R. LONG AND CHRISTOPHER ANTHONY

216

35. Methanol Defiydrogenase from Thermotolerant Methylotroph Bacillus CI N. ARFMAN AND L. DIJKHUIZEN

223

36. Methylamine Oxidase from Arthrobacter P1 WHilAM S. MCINTIRE 37. Methylamine Dehydrogenase from Thiobacillus versutus

227

JOHN E. VAN WIELINK, JOHANNES FRANK, AND JOHANNIS A. DUINE

235

38. Methylamine Dehydrogenases from Methylotrophic Bacteria VICTOR L. DAVIDSON

241

39. Methylamine Dehydrogenase from Methylobacillus flagellaturn MICHAEL Y. KIRIUKHIN, ANDREY Y. CHISTOSERDOV, AND YURI D. TSYGANKOV

247

WILLIAM S. MCINT1RE

7'50

40. Trimethylamine Dehydrogenase from Bacterium W3A1 41. Isolation, Preparation, and Assay of Pyrroloquinoline Quinone R. A. VAN DER MEER, B. W. GROEN, M. A. G. VAN KLEEF, J. FRANK, J. A. JONGEJAN, AND J. A. DUINE

260

42. Blue Copper Proteins Involved in Methanol and Methylamine Oxidation CHRISTOPHER ANTHONY

284

ALAN A. DISPIRITO

289

DARREN J. DAY AND CHRISTOPHER ANTHONY

298

43. Soluble Cytochromes c from Methylomonas A4 44. Soluble Cytochromes c of Methanol-Utilizing Bacteria

45. Cytochrome cL and Cytochrome CH from Hyphornicrobium X J. FRANK AND J. A. DUINE

303

46. Electron-Transfer Flavoproteins from Methylophilus methylotrophus and Bacterium W3AI MAZHARHUSAIN 47. Formaldehyde Dehydrogenases from Methylotrophs

309

MARGARET M. ATTWOOD

314

48. NAD-Linked, Factor-Independent, and Glutathione-Independent Aldehyde Dehydrogenase from Hyphomicrobium X J.A. DUINE 49. Formate Dehydrogenase from Methylosinus trichosporium OB3b

327

DAVID R. JOLLIE AND JOHN D. LIPSCOMB

331

50. Glucose-6-phosphate Dehydrogenase and 6-Phosphogluconate Dehydrogenase from Methylobacillus flagellatum LUDMILA V. KLETSOVA,MICHAELY. KIRIUKH1N. ANDREY Y. CHISTOSERDOV, AND YUR1 D. TSYGANKOV

335

lxiv

CONTENTS OF VOLUMES

168-174, 176-194

51. Glucose-6-phosphate Dehydrogenase and 6-Phosphogluconate Dehydrogenase from Arthrobacter globiformis A . P . SOKOLOV AND Y. A. TROTSENKO 52. Glucose-6-phosphate Dehydrogenase from Pseudomonas W6 DIETMAR MIETHE AND WOLFGANG BABEL 53. Citrate Synthases from Methylotrophs GABRIELE MI3LLER-KRAFT AND WOLFGANG BABEL 54. Dichloromethane Dehalogenase from Hyphomicrobium DM2 THOMAS LEISINGER AND DORIS KOHLER-STAUB

339 346 350 355

B. Assimilation of Formaldehyde in Methylotrophic Bacteria 55. Assay of Assimilatory Enzymes in Crude Extracts of Serine Pathway Methylotrophs PATRIOA M. GOODWIN 56. Serine Hydroxymethyltransferases from Methylobacterium organophilurn XX MARY E. LIDSTROM 57. Hydroxypyruvate Reductase from Methylobacteriurn extorquens AM1 CINDER KREMA AND MARY E. LIDSTROM 58. Malyl-CoA Lyase from Methylobacterium extorquens AMI A. J. HACKING AND J. R. QUAYLE 59. Synthesis of L-4-Malyl Coenzyme A PEGGY J. ARPS 60. 3-Hexulose-6-phosphate Synthase from Thermotolerant Methylotroph Bacillus C1 N. AREMAN, L. BYSTRYKH, N. I. GOVORUKHINA, AND L. DIJKHUIZEN 61. 3-Hexulose-6-phosphate Synthase from Mycobacterium gastri MBI9 NOBUO KATO 62. 3-Hexulose-6-phosphate Synthase from Acetobacter methanolicus MB58 ROLAND H. MULLER AND WOLFGANG BABEL 63. Transaldolase Isoenzymes from Arthrobacter P1 P. R. LEVERING AND L. DIJKHUIZEN

361 365 373 379 386 391 397 401 405

C. Methylotrophic Enzymes in Methanol-Utilizing Yeast 64. Cytochemical Staining Methods for Localization of Key Enzymes of Methanol Metabolism in Hansenula polymorpha MARTEN VEENHUIS AND IDA J. VAN DER KLEI 65. Alcohol Oxidase from Hansenula polymorpha CBS 4732 IDA J. VAN DER KLEI, LEONID V. BYSTRYKH, AND WIM HARDER 66. Amine Oxidases from Methylotrophic Yeasts PETER J. LARGE AND GEOFFREY W. HAYWOOD 67. Dihydroxyacetone Synthase from Candida boidinii KD1 LEONID V. BYSTRYKH, WIM DE KONING, AND WIM HARDER 68. Triokinase from Candida boidinii KDI LEONID V. BYSTRYKH, WIM DE KONING, AND WIM HARDER 69. Glycerone Kinase from Candida rnethylica KLAUS H. HOFMANN AND WOLFGANG BABEL 70. Formaldehyde Dehydrogenase from Methytotrophic Yeasts N O B U OKATO 71. Formate Dehydrogenase from Methylotrophic Yeasts N o a u o KATO 72. Catalase from Candida boidinii 2201 MITSUYOSHI UEDA, SABIHA MOZAFFAR, AND ATSUO TANAKA

411 420 427 435 445 451 455 459 463

CONTENTS OF VOLUMES 168--174, 176--194 VOLUME

lXV

189

RETINOIDS PART m M O L E C U L A R AND METABOLIC ASPECTS

S e c t i o n I. S t r u c t u r e a n d A n a l y s i s

A. Structure 1. Structure Characteristics of Natural and Synthetic Retinoids MICHAEL KLAUS 2. Synthetic Retinoic Acid Analogs: Handling and Characterization M. I. DAWSON AND P. D. HOBBS 3. Structural Characteristics of Synthetic Retinoids BRAHMA P. SANI AND DONALD L. HILL

3 15 43

B. Analysis 4. High-Performance Liquid Chromatography of Retinoids in Blood A. P. DE LEENHEER AND H. J. NELIS 5. High-Performance Liquid Chromatography of Retinoid Isomers: An Overview C. D. B. BRIDGES 6. High-Performance Liquid Chromatography of Retinoid Isomers GARY M. LANDERS 7. Separation of Fatty Acid Esters of Retinol by High-Performance Liquid Chromatography A. CATHARINE ROSS 8. Reversed-Phase High-Performance Liquid Chromatography of Retinyl Esters HAROLD C. FURR 9. Stable Isotope Dilution Mass Spectrometry to Assess Vitamin A Status ANDREW J. CLIFFORD, A. DANIEL JONES, AND HAROLD C. FURR 10. Mass Spectrometry of Methyl Ester of Retinoic Acid ANDRE P. DE LEENHEER AND WILLY E. LAMBERT 11. Characterization of Retinylidene Iminium Salts by High-Field IH and ~3C Nuclear Magnetic Resonance Spectroscopy GARY S. SHAW AND RONALD F. CHILDS 12. Resonance Raman and Infrared Difference Spectroscopy of Retinal Proteins FRIEDRICH SIEBERT 13. Analysis of Water-Soluble Compounds: Glucuronides ARUN B. BARUA 14. Determination of Retinoids in Plasma by High-Performance Liquid Chromatography and Automated Column Switching RONALD WYSS 15. Simultaneous, High-Performance Liquid Chromatographic Analysis of Retinol, Tocopherols, Lycopene, and a- and/3-Carotene in Serum and Plasma LAWRENCE A. KAPLAN, JUDITH m. MILLER, EVAN A. STEIN, AND MEIR J. STAMPFER 16. High-Performance Liquid Chromatography of Aromatic Retinoids and Isotretinoin in Biological Fluids H. BUN, N. R. AL-MALLAH, C. AUBERT, AND J. P. CANO 17. Determination of Retinol, a-Tocopherol, and /3-Carotene in Serum by Liquid Chromatography WILLIAM A. MACCREHAN 18. Separation of Retinyl Esters and Their Geometric Isomers by Isocratic Adsorption High-Performance Liquid Chromatography HANS K. BIESALSKI

50 60 70 81 85 94 104

112 123 136 146

[55

167 172 181

lxvi

CONTENTS OF VOLUMES 168-174, 176-194

S e c t i o n II. R e c e p t o r s , T r a n s p o r t , a n d B i n d i n g P r o t e i n s A. E x t r a c e l l u l a r 19. Purification and Properties of Plasma Retinol-Binding Protein WILLIAM S. BLANERAND DEWITT S. GOODMAN 20. Interstitial Retinol-Binding Protein: Purification, Characterization, Molecular Cloning, and Sequence SHAO-LING FONG AND C. D. B. BRIDGES 21. Purification and Assay of Interphotoreceptor Retinoid-Binding Protein from the Eye ALICE J. ADLER, GERALD J. CHADER, AND BARBARAWIGGERT

193 207 213

B. I n t r a c e l l u l a r 22. Identification of Receptors for Retinoids as Members of the Steroid and Thyroid Hormone Receptor Family VINCENTGIGUt~REAND RONALDM. EVANS 23. Retinoic Acid Acylation: Retinoylation

223

NORIKO TAKAHASHI AND THEODORE R. BREITMAN

233

24. Identification and Analysis of Retinoic Acid-Binding Proteins and Receptors from Nuclei of Mammalian Cells ANN K. DALY, CHRISTOPHER P. F. REDFERN, AND BERNARD MARTIN

239

25. Isolation and Binding Characteristics of Nuclear Retinoic Acid Receptors ANTON M. JETTEN, JOSEPH F. GRIPPO, AND CLARA NERVI

248

26. Nuclear Retinoic Acid Receptors: Cloning, Analysis, and Function MAGNUS PFAHL, MATY TZUKERMAN, XIAO-KUN ZHANG, JUERGEN M. LEHMANN, THOMAS HERMANN, KEN N. WILLS, AND GERHART GRAUPNER

256

C. S p e c i f i c M e t h o d s 27. Radioimmunoassays for Retinol-Binding Protein, Cellular Retinol-Binding Protein, and Cellular Retinoic Acid-Binding Protein WILLIAM S. BLANER 28. X-Ray Crystallographic Studies on Retinol-Binding Proteins

270

MARCIA NEWCOMER AND T. ALWYN JONES

281

29. In Situ Hybridization of Retinoid-Binding Protein Messenger RNA DIANNE ROBERT SOPRANO AND DEWITT S. GOODMAN

286

30. Gel Electrophoresis of Cellular Retinoic Acid-Binding Protein, Cellular RetinolBinding Protein, and Serum Retinol-Binding Protein GEORGES SIEGENTHALER

31. Cellular Retinoic Acid-Binding Protein from Neonatal Rat Skin: Purification and Analysis CHRISTOPHER P. F. REDFERN AND ANN K. DALY 32. Tissue Distribution of Cellular Retinol-Binding Protein and Cellular Retinoic Acid-Binding Protein: Use of Monospecific Antibodies for Immunohistochemistry and cRNA for in Situ Localization of mRNA CHRISTER BUSCH, PUSPHA SAKENA, KEIKO FUNA, HANS NORDLINDER, AND ULF ERIKSSON

299

307

315

33. Retinoids Bound to Interstitial Retinol-Binding Protein during the Visual Cycle ZHENG-SHI LIN, SHAO-LINGFONG, AND C. D. B. BRIDGES 34. Purification of Cellular Retinoic Acid-Binding Protein from Human Placenta

325

MICHIMASA KATO, MASATAKAOKUNO, AND YASUTOSHI MUTO

330

35. Differentiation of Embryonal Carcinoma Cells in Response to Retinoids PATRICIO ABARZ13AAND MICHAEL I. SHERMAN

337

lxvii

CONTENTS OF VOLUMES 168-174, 1 7 6 - 1 9 4

36. Parasite Retinoid-Binding Proteins BRAHMA P. SANI 37. Purification of Cellular Retinoic Acid-Binding Proteins Types I and II from Neonatal Rat Pups JOHN STUART BAILEY AND CHI-HUNG SIU 38. Purification of Adipocyte Lipid-Binding Protein from Human and Murine Cells VALERIE MATARESE, MELISSA K. BUELT, LAURIE L. CHINANDER, AND DAVID A. BERNLOHR 39. Binding of I l-cis-Retinaldehyde to Cellular Retinaldehyde-Binding Protein from Pigment Epithelium MARIA A. LIVREA AND LUISA TESORIERE

348 356

363 369

D. Membrane Interactions 40. Interactions of Retinoids with Phospholipid Membranes: Optical Spectroscopy WILLIAM STILLWELL AND STEPHEN R. WASSALL

41. Interactions of Retinoids with Phospholipid Membranes: Electron Spin Resonance STEPHEN R. WASSALL AND WILLIAM STILLWELE 42. Transfer of Retinol from Retinol-Binding Protein Complex to Liposomes and across Liposomal Membranes GI3RANFEX AND GUNVOR JOHANNESSON 43. Exchange of Retinoids between Lipid Vesicles and Rod Outer Segment Membranes WILLEM J. DE GRIP AND FRANS J. M. DAEMEN 44. Intermembranous Transfer of Retinoids ROBERT R. RANDO AND FAAN WEN BANGERTER

373 383

394 402 411

45. Incorporation of/]-Carotene into Mixed Micelles LOUISE M. CANFIELD, THOMAS A. FRITZ, AND THOMAS E. TARARA

S e c t i o n III. E n z y m o l o g y

418

and Metabolism

46. Carotenoid Conversions JAMES ALLEN OLSON AND M. R ~ LAKSHMAN 47. Carotenoid Cleavage: Alternative Pathways L. E. GERBER AND K. L. SIMPSON 48. Isoenzymes of Alcohol Dehydrogenase in Retinoid Metabolism XAVIER PARISS AND PERE JULIA

425 433 436

49. Measurement of Acyl Coenzyme A-Dependent Esterification of Retinol A. CATHARINE ROSS

442

MARK D. BALL

446

PAUL N. MACDONALD AND DAVID E. ONG

450

50. Acyl Coenzyme A-Dependent Retinol Esterification 51. Assay of Lecithin-Retinol Acyltransferase

52. Bile Salt-lndependent Retinyl Ester Hydrolase Activities Associated with Membranes of Rat Tissues EARL H. HARRISON AND JOSEPH L. NAPOLI 53. Quantification and Characteristics of Retinoid Synthesis from Retinol and /3Carotene in Tissue Fractions and Established Cell Lines JOSEPH L. NAPOLI 54. Retinol 4-Hydroxylase M . A . LEO AND C. S. LIEBER 55. Assay of Liver Retinyl Ester Hydrolase DALE A. COOPER 56. Assay of the Retinoid Isomerase System of the Eye PAUL S. BERNSTEIN AND ROBERT R. RANDO

57. Assay of all-trans---~ ll-cis-Retinoid Isomerase Activity in Bovine Retinal Pigment Epithelium MARIA A. LIVREA AND LUISA TESORIERE 58. Structure-Function Analyses of Mammalian Cellular Retinol-Binding Proteins by Expression in Escherichia coli MARC S. LEVlN, ELLEN L1, AND JEFFERY 1. GORDON

459

470 483 490 494

503

506

lxviii

CONTENTS OF VOLUMES 168-174, 176-194

59. NAD+-Dependent Retinol Dehydrogenase in Liver Microsomes M. A. LEO AND C. S. LIEBER 60. Metabolism of Retinoic Acid and Retinol by Intact Cells and Cell Extracts MARY LOU GUBLER AND MICHAEL I. SHERMAN 61. Retinoic Acid Formation from Retinol and Retinal Metabolism in Epidermal Cells GEORGES SIEGENTHALER

520 525 530

V O L U M E 190 R E T I N O I D S PART B C E L L D I F F E R E N T I A T I O N AND C L I N I C A L APPLICATIONS

Section I. Cell Differentiation A. Normal Cells 1. Analysis of the Visual Cycle by Short-Term Incubation of Isolated Retinal Pigment Epithelial Cells ADRIAN M. TIMMERS AND WILLEM J. DE GRIP 2. Vitamin A-Mediated Regulation of Keratinocyte Differentiation GEORGE J. GIUDICE AND ELAINE V. FUCHS 3. Retinoids and Lipid Changes in Keratinocytes MARIA PONEC AND ARIJ WEERHEIM 4. Down-Regulation of Squamous Cell-Specific Markers by Retinoids: Transglutaminase Type I and Cholesterol Sulfotransferase ANTON M. JETTEN, MARGARET A. GEORGE, AND JAMES I. REARICK 5. Isolation, Purification, and Characterization of Liver Cell Types HENK F. J. HENDRIKS, ADRIAAN BROUWER, AND DICK L. KNOOK 6. Isolation and Cultivation of Rat Liver Stellate Cells RUNE BLOMHOFF AND TROND BERG 7. Sertoli Cells of the Testis: Preparation of Cell Cultures and Effects of Retinoids ALICE F. KARL AND MICHAEL D. GRISWOLD 8. Measurement of Retinoid Effects on Epidermal Renewal LEONARD M. MILSTONE 9. Retinoids and Cell Adhesion LUIGI M. DE LUCA, SERGIO ADAMO, AND SHIGEMI KATO

3 18 30

42 49 58 71 76 81

10. Retinoids and Control of Epithelial Differentiation and Keratin Biosynthesis in Hamster Trachea LUIGI M. DE LUCA, FREESIA L. HUANG, AND DENNIS R. RooP

91

B. Cell Lines 11. Inhibition of Tumor Cell Growth by Retinoids REUBEN LOTAN, DAFNA LOTAN, AND PETER G. SACKS 12. Maintenance and Use of F9 Teratocarcinoma Cells ANDREW L. DARROW, RICHARD J. RICKLES, AND SIDNEY STRICKLAND 13. Growth and Differentiation of Human Myeloid Leukemia Cell Line HL60 THEODORE R. BRE1TMAN

100 110 118

CONTENTS OF VOLUMES 168-174, 176-194 14. Assays for Expression of Genes Regulated by Retinoic Acid in Murine Teratocarcinoma Cell Lines LORRAINE J. GUDAS 15. Retinoid-Binding Proteins in Retinoblastoma Cells SHAO-LING FONG AND C. D. B. BRIDGES 16. Retinoid-Binding Proteins in Embryonal Carcinoma Cells JOSEPH F. GRIPPO AND MICHAEL I. SHERMAN

lxix 131 141 148

C. Tissues and Organ Culture 17. Acyl-CoA:Retinol Acyltransferase and Lecithin:Retinol Acyltransferase Activities of Bovine Retinal Pigment Epithelial Microsomes JOHN C. SAARI AND D. LUC1LLE BREDBERG 18. High-Performance Liquid Chromatography of Natural and Synthetic Retinoids in Human Skin Samples ANDERS VAHLQUIST, HANS TORM~,, OLA ROLLMAN, AND EVA ANDERSSON 19. Retinoids and Rheumatoid Arthritis: Modulation of Extracellular Matrix by Controlling Expression of Collagenase CONSTANCE E. BRINCKERHOFF

156

163 175

D. Retinoids as Morphogens and Teratogens 20. Regenerating Limbs DAVID L. STOCUM AND MALCOLM MADEN 21. Targeted Slow-Release of Retinoids into Chick Embryos SARA WEDDEN, CHRISTINA THALLER, AND GREGOR EICHELE 22. Biosynthesis of 3,4-Didehydroretinol and Fatty Acyl Esters of Retinol and 3,4Didehydroretinol by Organ-Cultured Human Skin HANS T6RM~ AND ANDERS VAHLQUIST

189 201

210

E. Cell Synopsis 23. Retinoid-Sensitive Cells and Cell Lines

BRAD AMOS AND REUBEN LOTAN

217

Section II. Nutrition, Tissue and Immune Status, and Antioxidant Action A. Nutrition 24. Preparation of Vitamin A-Deficient Rats and Mice JOHN EDGAR SMITH 25. Use of Food Composition Tables for Retinol and Provitamin A Carotenoid Content KENNETH L. SIMPSON

229 237

B. Tissue and Immune Status 26. Biochemical and Histological Methodologies for Assessing Vitamin A Status in Human Populations BARaARA A. UNDERWOOD 27. Characterization of Immunomodulatory Activity of Retinoids DENISE A. FAHERTY AND ADRIANNE BENDICH 28. Immunotrophic Methodology KATHEEEN M. NAUSS, A. CATHARINE ROSS, AND SALLY S. TWINING

242 252 259

lxx

CONTENTS OF VOLUMES 168-174, 176-194 C. A n t i o x i d a n t A c t i o n

29. Antioxidant Activity of Retinoids MIDORIHIRAMATSUAND LESTERPACKER 30. Inhibition of Microsomal Lipid Peroxidation by 13-cis-Retinoic Acid

273

VICTOR M. SAMOKYSZYNAND LAWRENCE J. MARNETT

281

S e c t i o n III. P h a r m a c o k i n e t i c s , P h a r m a c o l o g y , a n d T o x i c o l o g y

A, P h a r m a c o k i n e t i c s 31. Retinoids: An Overview of Pharmacokinetics and Therapeutic Value H. GOLENICK, R. EHLERT, G. RINCK, AND C. E. ORFANOS 32. Experimental and Kinetic Methods for Studying Vitamin A. Dynamics in Vivo

291

MICHAEL H. GREEN AND JOANNE BALMER GREEN

304

33. Quantification of Embryonic Retinoic Acid Derived from Maternally Administered Retinol DEVENDRA M. KOCHHAR

317

B. P h a r m a c o l o g y 34. Effect of Retinoids on Sebaceous Glands STANLEY S. SHAPIRO AND JAMES HURLEY

326

35. Retinoid Effects on Sebocyte Proliferation THOMAS I. DORAN AND STANLEY S. SHAPIRO

334

36. Effects of Retinoids on Human Sebaceous Glands Isolated by Shearing TERENCE KEALEY

338

GERARD J. GENDIMENICO, ROBERT J. CAPETOLA, MARVIN E. ROSENTHALE, JOHN L. McGUIRE, AND JAMES A. MEZICK

346

37. Retinoid Modulation of Phorbol Ester Effects in Skin

38. Retinoid Effects on Photodamaged Skin GRAEME F. BRYCE AND STANLEY S. SHAPIRO

39. Effects of Topical Retinoids on Photoaged Skin as Measured by Optical Profilometry GARY L. GROVEAND MARYJo GROVE 40. Retinoid Effects on Photodamaged Skin LORRAINEH. KLIGMAN 41. Testing of Retinoids for Systemic and Topical Use in Human Psoriasis and Other Disorders of Keratinization: Mouse Papilloma Test KAMPE TEELMANN 42. Cancer Chemoprevention by Retinoids: Animal Models RICHARD C. MOON AND RAJENDRA G. MEHTA

352

360 372 382 395

C. T o x i c o l o g y 43. Utility of Disposition Data in Evaluating Retinoid Developmental Toxicity CALVIN C. WILLHITEAND STEVENA. BOOK 44. Teratogenicity of Retinoids: Mechanistic Studies

406

ANDREAS KISTLER AND W. BRIAN HOWARD

418

45. Testing of Retinoids for Teratogenicity in Vitro: Use of Micromass Limb Bud Cell Culture ANDREASKISTLERAND W. BRIANHOWARD 46. Testing of Retinoids for Teratogenicity in Vivo

427

ANDREAS KISTLER AND W. BRIAN HOWARD

433

47. Correlation of Transplacental and Maternal Pharmacokinetics of Retinoids during Organogenesis with Teratogenicity HEINZ NAU

437

CONTENTS OF VOLUMES 168-174, 176-194

lxxi

V O L U M E 191 BIOMEMBRANES PART V C E L L U L A R AND SUBCELLULAR TRANSPORT: EPITHELIAL CELLS 1. Epithelial Transport: An Introduction

KARl. JULIUS ULLRICH

2. Determination of Paracellular Shunt Conductance in Epithelia G. KOTTRA AND E. FROMTER

Section I. Kidney A. Intact Organ 3. Isolated Perfused and Nonfiltering Kidney PATRICIO SILVA 4. Multiple Indicator Dilution and the Kidney: Kinetics, Permeation. and Transport in Vivo CHARLES J. LUMSDEN AND MELVIN SILVERMAN 5. Micropuncture Techniques in Renal Research IHAB M. SHAFIK AND GARY A. QUAMME 6. Kidney: Microperfusion-Double-Perfused Tubule in Situ KARL JULIUS ULLRICH AND GERHARD RUMRICH

31 34 72 98

B. Glomerular Filtration 7. Transcapillary Fluid Transport in the Glomerulus AIDA YARED AND IEKUNI 1CHIKAWA 8. Preparation and Study of Isolated Glomeruli DETLEF SCHLONDORFF 9. Isolation and Study of Glomerular Cells D . A . TROVER AND J. I. KREISBERG 10. Isolation and Culture of Juxtaglomerular and Renomedullary Interstitial Cells E. ERIC MUIRHEAD, WILTON m. RIGHTSEL, JAMES A. PlTCOCK. AND TADASHI INAGAMI

107 131) 141

152

C. Isolated Tubules 11. Comparative Kidney Tubule Sources, Isolation, Perfusion, and Function KLAUS W. BEYENBACH AND WILLIAM H. DANTZLER 12. Microdissection of Kidney Tubule Segments P. A. WRIGHT, M. B. BURG, AND M. A. KNEPPER 13. Measurement of Transmural Water Flow in Isolated Perfused Tubule Segments JAMES C. WILLIAMS, JR., AND JAMES A. SCHAFER 14. Identification and Study of Specific Cell Types in Isolated Nephron Segments Using Fluorescent Dyes GEORGEJ. SCHWARTZ AND QAIS AL-AWQATI 15, Functional Morphology of Kidney Tubules and Cells in Situ BRIGITTE KAISSLING AND MICHEL LE HIR 16, An Electrophysiological Approach to the Study of Isolated Perfused Tubules RAINER GREGER 17. Hormonal Receptors in the Isolated Tubule FRAN(~OIS MOREL AND DANIEL BUTLEN 18, Metabolism of Isolated Kidney Tubule Segments GABRIELE WIRTHENSOHN AND WALTER G. GUDER 19. Endocytosis and Lysosomal Hydrolysis of Proteins in Proximal Tubules THOMAS MAACK AND C. HYUNG PARK

167 226 232 253 265 289 303 325 340

lxxii

CONTENTS OF VOLUMES 168-174, 176-194

20. Flux Measurements in Isolated Perfused Tubules JAMES A. SCHAFERAND JAMESC. WILLIAMS,JR. 21. Measurements of Volume and Shape Changes in Isolated Tubules WILLIAM B. GUGGINO,DIANE MARKAK1S,AND L. MARIOAMZEL

354 371

D. Isolated Renal Cells 22. Transport in Isolated Cells from Defined Nephron Segments ROLF K. H, KINNE 23. Primary Culture of Isolated Tubule Cells of Defined Segmental Origin

380

MICHAEL F, HORSTER AND MASAYOSHI SONE

409

24. Tissue Culture of Established Renal Cell Lines N . L . SIMMONS 25. Giant MDCK Cells: A Novel Expression System H. OBERLEITHNER, A. SCHWAB,H.-J. WESTPHALE,B. SCHURICHT, B. POSCHEL, AND a . KOEPSELL

426

437

E. Isolation of Membrane Vesicles 26. Isolation of Lumenal and Contralumenal Plasma Membrane Vesicles from Kidney EVAMARIAKINNE-SAFFgANAND ROLE K. H. KINNE 27. Transport Studies by Optical Methods G. SACHS, E. RABON, AND S. J. D. KARLISH 28. Stoichiometry of Coupled Transport Systems in Vesicles R. JAMESTURNER 29. Phosphate Transport in Established Renal Epithelial Cell Lines J. BIBER, K. MALMSTROM, S. RESHKIN, AND n . MURER 30. ATP-Driven Proton Transport in Vesicles from the Kidney Cortex

469 479

IVAN SABOLI(~AND GERHARD BURCKHARDT

505

450

494

F. Hormonal Modulation 31. Aldosterone 32. The Cellular Action of Antidiuretic Hormone

DIANA MARVER

520

DENNIS BROWN, ALAN VERKMAN, KARL SKORECKI, AND DENNIS AUSIELLO

551

G. R e c o n s t i t u t i o n o f T r a n s p o r t P r o t e i n s 33. Purification and Reconstitution of Epithelial Chloride Channels DONALD W. LANDRY, MYLES A. AKABAS, CHRISTOPHER REDHEAD, AND QAIS AL-AWQATI

572

34. Reconstitution and Fractionation of Renal Brush Border Transport Proteins HERMANN KOEPSELL AND STEFAN SEIBICKE

583

S e c t i o n II. S t i m u l u s S e c r e t i o n C o u p l i n g in E p i t h e l i a 35. Receptor Identification RAKESH VINAYEKAND JERRYD. GARDNER 36. cAMP Technologies, Functional Correlates in Gastric Parietal Cells

609

CATHERINE S. CHEW

640

37. Stimulus-Secretion Coupling: General Models and Specific Aspects in Epithelial Cells HOWARD RASMUSSEN

661

CONTENTS OF VOLUMES 168-174, 176-194

lxxiii

38. Metabolism and Function of Phosphatidylinositol-Derived Arachidonic Acid LOWELL E. HOKIN 39. Measurement of Intracellular Free Calcium to Investigate Receptor-Mediated Calcium Signaling CARL A. HANSEN, JONATHAN R. MONCK, AND JOHN R. WILLIAMSON 40. Two-Stage Analysis of Radiolabeled Inositol Phosphate Isomers K. A. WREGGETT, D. J. LANDER, AND R. F. IRVINE

S e c t i o n III.

676

691 707

P h a r m a c o l o g i c a l A g e n t s in E p i t h e l i a l T r a n s p o r t

41. Pharmacological Agents of Gastric Acid Secretion: Receptor Antagonists and Pump lnhibitors BJORN WALLMARKAND JAN FRYKLUND 42. Cation Transport Probes: The Amiloride Series THOMAS R. KLEYMAN AND EDWARD J. CRAGOE, JR. 43. Photoaffinity-Labeling Analogs of Phlorizin and Phloretin: Synthesis and Effects on Cell Membranes DONALD F. DIEDRICH 44. Diuretic Compounds Structurally Related to Furosemide SCOTT M. O'GRADY, MARK W. MUSCH, AND MICHAEL FIELD 45. Chloride Channel Blockers RAINER GRE6ER

721 739 755 781 793

Section IV. Targeting and Intracellular Trafficking in Epithelial Cells 46. In Vitro Recovery of Exocytic Transport Vesicles from Polarized MDCK Cells MARK K. BENNETT, ANGELA WANDINGER-NEss, ANDRI~ W. BR,/.NDLI, AND KAI SIMONS 47. Biogenesis of the Rat Hepatocyte Plasma Membrane JAMES R. BARTLES AND ANN L. HUBBARD 48. Oligomerization and Protein Transport ROBERT W. DOMS

813 825 841

VOLUME 192 BIOMEMBRANES PART W C E L L U L A R AND SUBCELLULAR TRANSPORTZ EPITHELIAL CELLS

Section I. Gastrointestinal System A. Salivary Gland 1. Salivary Secretion: Studies on Intact Glands in Vivo and in Vitro I. NOVAK 2. Determination of Short-Circuit Current in Small Tubular Structures via Cable Analysis K. HAAG AND H. KNAUF 3. Dispersed Salivary Gland Acinar Cell Preparations for Use in Studies of Neuroreceptor-Coupled Secretory Events BRUCE J. BAUM, INDUS. AMBUDKAR, JOSEPH HELMAN, VALERIE J. HORN. JAMES E. MELVIN, LAWRENCE M. MERTZ, AND R. JAMES TURNER

3 15

26

lxxiv

CONTENTS OF VOLUMES

168-174, 176-194

B. Stomach 4. Intraceilular Ion Activities and Membrane Transport in Parietal Cells Measured with Fluorescent Dyes PAUL A. NEGULESCU AND TERRY E. MACHEN 5. Electrophysiological Techniques in the Analysis of Ion Transport across Gastric Mucosa S. CURCI AND E. FROMTER 6. Gastric Glands and Cells: Preparation and in Vitro Methods THOMAS BERGLINDH 7. Permeabilizing Parietal Ceils DANUTA H. MALINOWSKA 8. Pepsinogen Secretion in Vitro STEPHEN HERSEY, LAURA TANG, JAN POHL, AND MELISSA MILLER 9. HCOi Secretion: Stomach, Duodenum GUNNAR FLEMSTROM AND HENRIK FORSSELL 10. Isolation of H+,K+-ATPase-Containing Membranes from the Gastric Oxyntic Cell W . W . REENSTRA AND J. G. FORTE 11. Peptic Granules from the Stomach BRIAN E. PEERCE 12. Isolation and Primary Culture of Endocrine Cells from Canine Gastric Mucosa TADATAKA YAMADA

38 82 93 108 124 139 151 165 176

C. Pancreas 13. Isolation of Pancreatic Islets and Primary Culture of the Intact Microorgans or of Dispersed Islet Cells CLAES B. WOLLHEIM, PAOLO MEDA, AND PHILIPPE A. HALBAN 14. Establishment and Culture of Insulin-Secreting//Cell Lines CLAES B. WOLLHEIM, PAOLO MEDA, AND PHILIPPE A. HALBAN 15. Membrane Potential Measurements in Pancreatic//Cells with Intracellular Microelectrodes HANS PETER MEiSSNER 16. Stimulation of Secretion by Secretagogues STEPHEN A. WANK, ROBERT T. JENSEN, AND JERRY D. GARDNER 17. Pancreatic Secretion: In Vivo, Peffused Gland, and Isolated Duct Studies R. M. CASE AND B. E. ARGENT 18. Dispersed Pancreatic Acinar Cells and Pancreatic Acini DELLA MENOZZI, ROBERT T. JENSEN, AND JERRY D. GARDNER 19. Permeabilizing Cells: Some Methods and Applications for the Study of Intracellular Processes IRENE SCHULZ 20. Electrophysiology of Pancreatic Acinar Cells O. H. PETERSEN, M. WAKUI, Y. OSlPCHUK, D. YULE, AND D. V. GALLACHER

188 223 235 247 256 271 280 300

D. Intestine 21. Characterization of a Membrane Potassium Ion Conductance in Intestinal Secretory Cells Using Whole Cell Patch-Clamp and Calcium Ion-Sensitive Dye Techniques MICHAEL E. DUFFEY, DANIEL C. DEVOR, ZAHUR AHMED, AND STEVEN M. SIMASKO 22. Isolation of Intestinal Epithelial Cells and Evaluation of Transport Functions GEORGE A. KIMMICH 23. Isolation of Enterocyte Membranes AUSTIN K. MIRCHEFF AND EMILE J. J. M. VAN CORVEN

309 324 341

CONTENTS OF VOLUMES 168--174, 176--194 24. Established Intestinal Cell Lines as Model Systems for Electrolyte Transport Studies KIERTISIN DHARMSATHAPHORN AND JAMES L. MADARA 25. Sodium Chloride Transport Pathways in Intestinal Membrane Vesicles ULRICH HOPFER 26. Advantages and Limitations of Vesicles for the Characterization and the Kinetic Analysis of Transport Systems A. BERTELOOT AND G. SEMENZA 27. Isolation and Reconstitution of the Sodium-Dependent Glucose Transporter PARAMESWARAMALATHI AND MARK TAKAHASHI 28. Calcium Transport by Intestinal Epithelial Cell Basolateral Membrane JULIAN R. F. WALTERS AND MILTON M. WEISER 29. Electrical Measurements in Large Intestine (Including Caecum, Colon, Rectum) ULRICH HEGEL AND MICHAEL FROMM

IXXV

354 389 409 438 448 459

E. Liver 30. The Use of Isolated Perfused Liver in Studies of Biological Transport Processes MURAD OOKHTENS AND NEIL KAPLOWITZ 31. Measurement of Unidirectional Calcium Ion Fluxes in Liver LAURENT COMBETTES, CATHERINE DARGEMONT, JEAN-PIERRE MAUGER, AND MICHEL CLARET 32. Preparation and Specific Applications of Isolated Hepatocyte Couplets J. L. BOYER, J. M. PHILLIPS, AND J. GRAF 33. Characterizing Mechanisms of Hepatic Bile Acid Transport Utilizing Isolated Membrane Vesicles JAMES L. BOYER AND PETER J. MEIER 34. Preparation of Basolateral (Sinusoidal) and Canalicular Plasma Membrane Vesicles for the Study of Hepatic Transport Processes PETER J. MEIER AND JAMES L. BOYER

485

495 501 517

534

Section II. Other Epithelia A. Trachea 35. Electrogenic and Electroneutral Ion Transporters and Their Regulation in Tracheal Epithelium CAROLE M. LIEDTKE 36. Transformation of Airway Epithelial Cells with Persistence of Cystic Fibrosis or Normal Ion Transport Phenotypes JAMES R. YANKASKASAND RICHARD C. BOUCHER

549

565

B. Cornea and Eye 37. Cell Culture of Bovine Corneal Endothelial Cells and Its Application to Transport Studies MICHAEL WIEDERHOLT AND THOMAS J. JENTSCH

571

C. Sweat Glands 38. Methods for Studying Eccrine Sweat Gland Function in Vivo and in Vitro KENZO SATO AND FUSAKO SATO

583

lxxvi

CONTENTS OF VOLUMES 168-174, 176--194

D. Insect Epithelia 39. Isolation, Voltage Clamping, and Flux Measurements in Lepidopteran Midgut WILLIAM R. HARVEY, DWIGHT N. CRAWFORD, AND DANIEL D. SPAETH 40. Isolation of Goblet Cell Apical Membrane from Tobacco Hornworm Midgut and Purification of Its Vacuolar-Type ATPase HELMUT WIECZOREK, MOIRA CIOFFI, ULLA KLEIN, WILLIAM R. HARVEY, HELMUT SCHWEIKL, AND MICHAEL G. WOLFERSBERGER 41. Methods for the Study of Fluid and Solute Transport and Their Control in Insect Malpighian Tubules S . H . P . MADDRELL AND J. A. OVERTON

599

608 617

E. Urinary Bladder 42. Physiological Approaches for Studying Mammalian Urinary Bladder Epithelium SIMON A. LEWIS AND JOHN W. HANRAHAN

632

F. Gallbladder 43. Electrophysiological Methods for Studying Ion and Water Transport in Necturus Gall Bladder Epithelium G. ALTENRERG, J. COPELLO, C. COTTON, K. DAWSON, Y. SEGAL, F. WEHNER, AND L. REUSS

650

G. Amphibian and Reptilian Epithelia 44. Toad Urinary Bladder as a Model for Studying Transepithelial Sodium Transport MORTIMER M. CIVAN AND HAIM GARTY 45. Methods to Detect, Quantify, and Minimize Edge Leaks in Ussing Chambers L. G. M. GORDON, G. KOTTRA, AND E. FROMTER 46. Amphibian Nephron: Isolated Kidney and Cell Fusion H. OBERLEITHNER, B. GASSNER, P. DIETL, AND W. WANG 47. Turtle Colon: Keeping Track of Transporters in the Apical and Basolateral Membranes DAVID C. DAWSON AND DEAN CHANG

683 697 710 734

H. Fish Epithelia 48. Ion Transport of Marine Teleost Intestine MARK W. MUSCH, SCOTT M. O'GRADY, AND MICHAEL FIELD 49. Shark Rectal Gland PATRICIO SILVA, RICHARD J. SOLOMON, AND FRANKLIN H, EPSTEIN

746 754

VOLUME 193 MASS SPECTROMETRY

S e c t i o n I. G e n e r a l T e c h n i q u e s 1. Methods of Ionization 2. Mass Analyzers

ALEX G. HARRISON AND ROBERT J. COTTER KEITH R. JENNINGS AND GREGORY G. DOLNIKOWSKI

3 37

CONTENTS OF VOLUMES 168--174, 176--194 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. 14. 15. 16. 17.

Ixxvii

Detectors SYD EVANS Selected-Ion Measurements J. THROCK WATSON Liquid Chromatography-Mass Spectrometry MARVIN L. VESTAL Tandem Mass Spectrometry: Multisector Magnetic Instruments MICHAEL L. GROSS Tandem Mass Spectrometry: Quadrupole and Hybrid Instruments RICHARD A. YOST AND ROBERT K. BOYD Matrix Selection for Liquid Secondary Ion and Fast Atom Bombardment Mass Spectrometry EDWIN DE PAUW Continuous-Flow Fast Atom Bombardment Mass Spectrometry RICHARD M. CAPRIOLI AND WILLIAM Z. MOORE Collision-Induced Dissociation ROGERN. HAYES AND MICHAEL L. GROSS Principles of Californium-252 Plasma Desorption Mass Spectrometry Applied to Protein Analysis RONALD D. MACFARLANE Mass Spectrometry of Peptides and Proteins by Matrix-Assisted Ultraviolet Laser Desorption/Ionization FRANZ HILLENKAMP AND MICHAEL KARAS Utility of Exact Mass Measurements KLAUS BIEMANN Selection and Use of Mass Reference Standards RICHARD M. MILBERG Chemical Derivatization for Mass Spectrometry DANIEL R. KNAPP Introduction of Deuterium by Exchange for Measurement by Mass Spectrometry JAMES A. McCLOSKEY Preparation of Labeled Molecules by Exchange with Oxygen-18 Water ROBERT C. MURPHY AND KEITH L. CLAY

61 86 107 131 154 201 214 237 263 280 295 305 314 329 338

S e c t i o n II. P e p t i d e s a n d P r o t e i n s 18. Peptides and Proteins: Overview and Strategy KLAUS BIEMANN 19. Enzymatic and Chemical Digestion of Proteins for Mass Spectrometry TERRY D. LEE AND JOHN E. SHIVELY 20. Strategies for Locating Disulfide Bonds in Proteins DAVID L. SMITH AND ZHONGRUI ZHOU 21. Reversed-Phase High-Performance Liquid Chromatography for Fractionation of Enzymatic Digests and Chemical Cleavage Products of Proteins KATHRYN L. STONE, JAMES I. ELLIOTT, GLENN PETERSON, WALTER McMURRAY, AND KENNETH R. WILLIAMS 22. Electrospray Ionization Mass Spectrometry CHARLES G. EDMONDS AND RICHARD D. SMITH 23. Sample Preparation for Plasma Desorption Mass Spectrometry PETER ROEPSTOREF 24. Molecular Weight Analysis of Proteins IAN JARDINE 25. Sequencing of Peptides by Tandem Mass Spectrometry and High-Energy Collision-Induced Dissociation KLAUS BIEMANN 26. Liquid Secondary Ion Mass Spectrometry of Phosphorylated and Sulfated Peptides and Proteins BRADFORD W. GIBSON AND PHILIP COHEN 27. Identification of Attachment Sites and Structural Classes of Asparagine-Linked Carbohydrates in Glycoproteins STEVEN A. CARR, JOHN R. BARR, GERALD D. ROBERTS, KALYAN R. ANUMULA, AND PAUL B. TAYLOR 28. Characterization of Recombinant Proteins HUBERT A. SCOBLE AND STEPHEN A. MARTIN

351 361 374

389 412 432 441 455 480

50[ 519

lxxviii

CONTENTS OF VOLUMES 168-174, 176-194

Section III. Glycoconjugates 29. 30. 31. 32. 33.

34. 35. 36. 37.

38. 39. 40.

Glycoconjugates: Overview and Strategy ROGER A. LAINE Linkage Analysis Using Lindberg Method CARL G. HELLERQVIST Linkage Analysis Using Reductive Cleavage Method GARY R. GRAY Linkage Positions in Glycoconjugates by Periodate Oxidation and Fast Atom Bombardment Mass Spectrometry ANNE-SOPHIEANGEL AND BO NILSSON Tandem Mass Spectrometry in Structural Characterization of Oligosaccharide Residues in Glycoconjugates WILHELM J. RICHTER, DIETER R. M()LLER, AND BRUNO DOMON Mass Spectrometry of Mixtures of Intact Glycosphingolipids Bo E. SAMUELSSON,WESTON PIMLOTT, AND KARL-ANDERSKARLSSON Preparation of Desorption Mass Spectrometry of Permethyl and Peracetyl Derivatives of Oligosaccharides ANNE DELL Desorption Mass Spectrometry of Oligosaccharides Coupled with Hydrophobic Chromophores LINDA POULTER AND A. L. BURLINGAME Oligosaccharide Characterization with High-Energy Collision-Induced Dissociation Mass Spectrometry BETH L. GILLECE-CASTROAND A. L. BURLINGAME Desorption Mass Spectrometry of Glycosphingolipids JASNA PETER-KATALINIt~AND HEINZ EGGE High-Mass Gas Chromatography-Mass Spectrometry of Permethylated Oligosaccharides GUNNAR C. HANSSONAND HASSE KARLSSON Tandem Mass Spectrometry of Glycolipids CATHERINE E. COSTELLOAND JAMES E. VATH

539 554 573 587

607 623 647 661

689 713 733 738

Section IV. Nucleic Acid Constituents 41. Constituents of Nucleic Acids: Overview and Strategy JAMESA. MCCLOSKEY 42. Preparation and Enzymatic Hydrolysis of DNA and RNA for Mass Spectrometry PAMELA F. CRAIN 43. Preparation of Trimethylsilyl Derivatives of Nucleic Acid Components for Analysis by Mass Spectrometry KARL H. SCHRAM 44. Analysis of RNA Hydrolyzates by Liquid Chromatography-Mass Spectrometry STEVEN C. POMERANTZAND JAMES A. McCLOSKEY 45. Electron Ionization Mass Spectra of Trimethylsilyl Derivatives of Nucleosides JAMES A. McCLOSKEY 46. Gas Chromatography-Mass Spectrometry of Free Radical-Induced Products of Pyrimidines and Purines in DNA MIRAL DIZDAROGLU 47. Analysis of 5-Methylcytosine in DNA by Isotope Dilution Gas ChromatographyMass Spectrometry PAMELA F. CRAIN

771 782 791 796 825 842 857

Appendixes 1. 2. 3. 4. 5. 6.

Mass and Abundance Values JAMES A. McCLOSKEY Reference Ions for Exact Mass Measurements RICHARD M. MILBERG Mass Spectra of Matrix Materials C . E . COSTELLO Calculation of Isotopic Abundance Distributions JAMES A. McCLOSKEY Nomenclature for Peptide Fragment Ions (Positive Ions) KLAUSBIEMANN Mass Values for Amino Acid Residues in Peptides KLAUS BIEMANN

869 870 875 882 886 888

CONTENTS OF VOLUMES 168-174, 176-194

lxxix

V O L U M E 194 GUIDE TO YEAST GENETICS AND MOLECULAR BIOLOGY

S e c t i o n I. B a s ic M e t h o d s of Y e a s t G e n e t i c s 1. 2. 3. 4.

Getting Started with Yeast FRED SHERMAN Micromanipulation and Dissection of Asci FRED SHERMANAND JAMES HICKS Mapping Yeast Genes FRED SHERMAN AND PAUL WAKEM Positional Mapping of Genes by Chromosome Blotting and Chromosome Fragmentation SANDRAL. GERRING, CARLA CONNELLY, AND PHILIP HIETER 5. Assay of Yeast Mating Reaction GEORGE F. SPRAGUE, JR. 6. Monitoring Meiosis and Sporulation in Saccharomyces cerevisiae YONA KASSIR AND GIORA SIMCHEN

3 21 38 57

77 94

7. Identifying Sporulation Genes, Visualizing Synaptonemal Complexes, and Large-Scale Spore and Spore Wall Purification ROCHELLE EASTON ESPOSITO, MICHAEL DRESSER, AND MICHAEL BREITENBACH

110

8. Putting the HO Gene to Work: Practical Uses for Mating-Type Switching IRA HERSKOWITZ AND ROBERT E. JENSEN

132

BETH ROCKMILL, ERIC J. LAMBIE, AND G. SHIRLEEN ROEDER

146

9. Spore Enrichment 10. Analysis and Manipulation of Yeast Mitochondrial Genes THOMAS D. FOX, LINDA S. FOLLEY, JULIO J. MULERO, THOMAS W. McMULLIN, PETER E. THORSNESS, LARS O. HEDIN, AND MARIA C. COSTANZO

149

S e c t i o n II. C l o n i n g a n d R e c o m b i n a n t D N A 11. DNA of Saccharomyces cereuisiae PETER PHILIPPSEN, AGATHE STOTZ, AND CHRISTINE SCHERF

169

12. High-Efficiency Transformation of Yeast by Electroporation DANIEL M. DECKER AND LEONARD GUARENTE

13. Transmission of Plasmid DNA to Yeast by Conjugation with Bacteria JACK A. HEINEMANN AND GEORGE F. SPRAGUE, JR. 14. Cloning Genes by Complementation in Yeast MARK D. ROSE AND JAMESR. BROACH 15. Gene Isolation with hgtl 1 System

182 187

195

RICHARD A. YOUNG AND RONALD W. DAVIS

230

16. Gene Overexpression in Studies of Saccharomyces cerevisiae JASPERRINE 17. Preparation of Clone Libraries in Yeast Artificial-Chromosome Vectors DAVID T. BURKEAND MAYNARDV. OLSON

239 251

S e c t i o n III. M a k i n g M u t a n t s 18. Classical Mutagenesis Techniques CHRISTOPHERW. LAWRENCE 19. Targeting, Disruption, Replacement, and Allele Rescue: Integrative DNA Transformation in Yeast RODNEY ROTHSTEIN

273 281

IXXX

CONTENTS OF VOLUMES 168-174, 176-194

20. In Vitro Mutagenesis and Plasmid Shuffling: From Cloned Gene to Mutant Yeast ROBERT S. SIKORSKIAND JEF D. BOEKE 21. Recovery of Plasmids from Yeast into Escherichia coli: Shuttle Vectors JEFFREY N. STRATHERNAND DAVIDR. HIGGINS 22. Shuttle Mutagenesis: Bacterial Transposons for Genetic Manipulations in Yeast

302 319

MERL W. HOEKSTRA, n . STEVEN SEIFERT, JAC NICKOLOFF, AND

FRED HEEFRON

329

DAVID J. GARFINKEL AND JEFFREY N. STRATHERN

342

23. Ty Mutagenesis in Saccharomyces cerevisiae 24. Transformation of Yeast Directly with Synthetic Oligonucleotides RICHARD P. MOERSCHELL, GOUTAM DAS, AND FRED SHERMAN

362

S e c t i o n IV. B i o c h e m i s t r y of G e n e E x p r e s s i o n 25, Vectors for Expression of Cloned Genes in Yeast: Regulation, Overproduction, and Underproduction JANE C. SCHNEIDER AND LEONARD GUARENTE 26. Vectors for Constitutive and Inducible Gene Expression in Yeast

373

MARK SCHENA, DIDIER PICARD, AND KEITH R. YAMAMOTO

389

27. Preparation of High Molecular Weight RNA KARL KOHRER AND HORST DOMDEY

398

28. Preparation and Analysis of Low Molecular Weight RNAs and Small Ribonucleoproteins Jo ANN WISE 29. Measurement of mRNA Decay Rates in Saccharomyces cerevisiae

405

ROY PARKER, DAVID HERRICK, STUART W. PELTZ, AND ALLAN JACOBSON

415

30. Labeling of RNA and Phosphoproteins in Saccharomyces cerevisiae JONATHAN R. WARNER

423

31. Tackling the Protease Problem in Saccharomyces cerevisiae ELIZABETH W. JONES 32. Structural and Functional Analysis of Yeast Ribosomal Proteins

428

H. A. RAUI~, W. H. MAGER, AND R. J. PLANTA

453

33. High-Expression Vectors with Multiple Cloning Sites for Construction of trpE Fusion Genes: pATH Vectors T. J. KOERNER, JOHN E. HILL, ALAN M. MYERS, AND ALEXANDER TZAGOLOFF

477

34. Production of Proteins by Secretion from Yeast DONALD T. MOIR AND LANCE S. DAVIDOW

491

35. Epitope Tagging and Protein Surveillance PETER A. KOLODZIFAAND RICHARD A. YOUNG

36. Reverse Biochemistry: Methods and Applications for Synthesizing Yeast Proteins in Vitro KEVIN STRUHL 37. In Vitro Protein Synthesis MICHAEL J. LEIBOWITZ, FRANCIS P. BARBONE, AND

508

520

DENISE E. GEORGOPOULOS

536

NEAL F. LUE, PETER M. FLANAGAN, RAYMOND J. KELLEHER III, ALED M, EDWARDS, AND ROGER D. KORNBERG

54.5

38. RNA Polymerase II Transcription in Vitro

39. Direct Sequence and Footprint Analysis of Yeast DNA by Primer Extension JON M. HUIBREGTSE AND DAVID R. ENGELKE

550

CONTENTS OF VOLUMES 168-174, 176-194

lxxxi

Section V. Cell Biology 40. Immunofluorescence Methods for Yeast JOHN R. PRINGLE, ALISON E. M. ADAMS, DAVID G. DRUB1N, AND BRIAN K. HAARER 41. Preparation of Yeast Cells for Thin-Section Electron Microscopy BRECK DYERS AND LORETTA GOETSCH 42. Immunogold Labeling of Yeast Ultrathin Sections MICHAEL W. CLARK 43. Analysis of Mitochondrial Function and Assembly MICHAEL P. YAFFE 44. Methods for Studying the Yeast Vacuole CHRISTOPHER J. ROBERTS, CHRISTOPHER K. RAYMOND, CARL T. YAMASHIRO, AND TOM H. STEVENS 45. Analysis of Polypeptide Transit through Yeast Secretory Pathway ALEX FRANZUSOFF, JONATHAN ROTHBLATT, AND RANDY SCHEKMAN 46. In Vitro Protein Translocation across Microsomal Membranes of Saccharomyces cere~isiae PABLO D. GARCIA, WILLIAM HANSEN, AND PETER WALTER 47. Analysis of Glycoproteins from Saccharomyces cerevisiae P. ORLEAN, M. J. KURANDA, AND C. F. ALBRIGHT 48. Yeast Endocytosis Assays VJEKOSLAV DULIC, MARK EGERTON, IBRAHIM ELGUIND1, SUSAN RATHS. BIRGIT SINGER, AND HOWARD RIEZMAN 49. Inducing and Assaying Heat-Shock Response in Saccharomyces cerevisiae CHARLES M. NICOLET AND ELIZABETH A. CRAIG 50. Nucleolar-Specific Positive Stains for Optical and Electron Microscopy MICHAEl_ W, CLARK

565 602 608 627

644 662 675 682

697 710 717

51. Staining of Actin with Fluorochrome-Conjugated Phalloidin ALISON E. M. ADAMS AND JOHN R. PRINGLE 52. Staining of Bud Scars and Other Cell Wall Chitin with Calcofluor JOHN R. PRINGLE 53. Isolation of Yeast Nuclei JOHN P. ARIS AND G1ANTER BLOBEL 54. Analysis of Chromosome Segregation in Saccharomyces cerevisiae JAMES H. SHERO, MICHAEL KOVAL, FORREST SPENCER, ROBERT E. PALMER, PHILIP HIETER, AND DOUGLAS KOSHLAND 55. Genetic Screens and Selections for Cell and Nuclear Fusion Mutants VIVIAN BERLIN, JUL1E A. DRILL, JOSHUA TRUEHEART, JEF D. BOEKE, AND GERALD R. FINK

729 732 735

749

774

S e c t i o n VI. Fission Y e a s t 56. Molecular Genetic Analysis of Fission Yeast Schizosaccharomyces pombe SERGIO MORENO, AMAR KLAR, AND PAUL NURSE

795

Section VII. Appendix 57. Genetic and Physical Maps of Saccharomyces cerevisiae ROBERT K. MORTIMER, DAVID SCHILD, C. REBECCA CONTOPOULOU, AND JONATHAN A. KANS

827

METHODS IN ENZYMOLOGY VOLUME I. Preparation and Assay of Enzymes

Edited by SIDNEY P. COLOWICK AND NATHAN O. KAPLAN VOLUME II. Preparation and Assay of Enzymes Edited by SIDNEY P. COLOWICK AND NATHAN O. KAPLAN VOLUME III. Preparation and Assay of Substrates Edited by SIDNEY P. COLOWICK AND NATHAN O. KAPLAN VOLUME IV. Special Techniques for the Enzymologist Edited by SIDNEY P. COLOWICK AND NATHAN O. KAPLAN VOLUME V. Preparation and Assay of Enzymes Edited by SIDNEY P. COLOWICK AND NATHAN O. KAPLAN VOLUME VI. Preparation and Assay of Enzymes (Continued) Preparation and Assay of Substrates Special Techniques

Edited by SIDNEY P. COLOWICK AND NATHAN O. KAPLAN VOLUME VII. Cumulative Subject Index Edited by SIDNEY P. COLOWICK AND NATHAN O. KAPEAN VOLUME VIII, Complex Carbohydrates Edited by ELIZABETH F. NEUFELD AND VICTOR GINSBURG VOLUME IX.

Carbohydrate Metabolism

Edited by WILLIS A. WOOD VOLUME X. Oxidation and Phosphorylation Edited by RONALD W. ESTABROOKAND MAYNARD E. PULLMAN VOLUME XI.

EnzymeStructure

Edited by C. H. W. HIRS VOLUME XII. Nucleic Acids (Parts A and B) Edited by LAWRENCE GROSSMAN AND KIVIE MOLDAVE VOLUME XIII. Citric Acid Cycle Edited by J. M. LOWENSTEIN VOLUME XIV. Lipids Edited by J. M. LOWENSTEIN

VOLUME XV. Steroids and Terpenoids Edited by RAYMOND B. CLAYTON VOLUME XVI. Fast Reactions

Edited by KENNETH KUSTIN VOLUME XVII. Metabolism of Amino Acids and Amines (Parts A and B) Edited by HERBERT TABOR AND CELIA WHITE TABOR lxxxiii

lxxxiv

METHODS IN ENZYMOLOGY

VOLUME XVIII. Vitamins and Coenzymes (Parts A, B, and C)

Edited by DONALD B. MCCORMICKAND LEMUEL D. WRIGHT VOLUME XIX. Proteolytic Enzymes

Edited by GERTRUDE E. PERLMANNAND LASZLOLORAND VOLUME XX. Nucleic Acids and Protein Synthesis (Part C)

Edited by KIVIE MOLDAVEAND LAWRENCEGROSSMAN VOLUME XXI. Nucleic Acids (Part D)

Edited by LAWRENCEGROSSMANAND KIVIE MOLDAVE VOLUME XXII. Enzyme Purification and Related Techniques

Edited by WILLIAMB. JAKOBY VOLUME XXIII. Photosynthesis (Part A)

Edited by ANTHONY SAN PIETRO VOLUME XXIV. Photosynthesis and Nitrogen Fixation (Part B)

Edited by ANTHONY SAN PIETRO VOLUME XXV. Enzyme Structure (Part B)

Edited by C. H. W. HIRS AND SERGE N. TIMASHEEF VOLUME XXVI. Enzyme Structure (Part C)

Edited by C. H. W. HIRS AND SERGE N. TIMASHEFF VOLUME XXVII. Enzyme Structure (Part D)

Edited by C. H. W. HIRS AND SERGE N. TIMASHEFF VOLUME XXVIII. Complex Carbohydrates (Part B)

Edited by VICTORGINSBURG VOLUME XXIX. Nucleic Acids and Protein Synthesis (Part E)

Edited by LAWRENCEGROSSMANAND KIVIE MOLDAVE VOLUME XXX. Nucleic Acids and Protein Synthesis (Part F)

Edited by KIVlE MOLDAVEAND LAWRENCEGROSSMAN VOLUME XXXI. Biomembranes (Part A)

Edited by SIDNEY FLEISCHERAND LESTER PACKER VOLUME XXXII. Biomembranes (Part B)

Edited by SIDNEY FLEISCHERAND LESTER PACKER VOLUME XXXIII. Cumulative Subject Index Volumes I-XXX

Edited by MARTHA G. DENNIS AND EDWARDA. DENNIS VOLUME XXXIV. Affinity Techniques (Enzyme Purification: Part B)

Edited by WILLIAM B. JAKOBYAND MEIR WILCHEK VOLUME XXXV. Lipids (Part B)

Edited by JOHN M. LOWENSTEIN VOLUME XXXVI. Hormone Action (Part A: Steroid Hormones)

Edited by BERT W. O'MALLEY AND JOEL G. HARDMAN

METHODS IN ENZYMOLOGY

IXXXV

VOLUME XXXVII. Hormone Action (Part B: Peptide Hormones)

Edited by BERT W. O'MALLEY AND JOEL G. HARDMAN VOLUME XXXVIII. Hormone Action (Part C: Cyclic Nucleotides)

Edited by JOEL G. HARDMAN AND BERT W. O'MALLEY VOLUME XXXIX. Hormone Action (Part D: Isolated Cells, Tissues, and Organ Systems) Edited by JOEL G. HARDMAN AND BERT W. O'MALLEY VOLUME XL. Hormone Action (Part E: Nuclear Structure and Function) Edited by BERT W. O'MALLEY AND JOEL G. HARDMAN VOLUME XLI. Carbohydrate Metabolism (Part B)

Edited by W. A. WOOD VOLUME XLII. Carbohydrate Metabolism (Part C)

Edited by W. A. WOOD VOLUME XLIII. Antibiotics

Edited by JOHN H. HASH VOLUME XLIV. Immobilized Enzymes

Edited by KLAUS MOSBACH VOLUME XLV. Proteolytic Enzymes (Part B)

Edited by LASZLO LORAND VOLUME XLVI. Affinity Labeling

Edited by WILLIAM B. JAKOBY AND MEIR WILCHEK VOLUME XLVII. Enzyme Structure (Part E)

Edited by C. H. W. HIRS AND SERGE N. TIMASHEFE VOLUME XLVIII. Enzyme Structure (Part F)

Edited by C. H. W. HIRS AND SERGE N. TIMASHEFF VOLUME XLIX. Enzyme Structure (Part G)

Edited by C. H. W. HIRS AND SERGE N. TIMASHEFF VOLUME L. Complex Carbohydrates (Part C)

Edited by VICTOR GINSBURG VOLUME LI. Purine and Pyrimidine Nucleotide Metabolism

Edited by PATRICIA A. HOFFEE AND MARY ELLEN JONES VOLUME LII. Biomembranes (Part C: Biological Oxidations)

Edited by SIDNEY FLE1SCHER AND LESTER PACKER VOLUME LIII. Biomembranes (Part D: Biological Oxidations)

Edited by SIDNEY FLEISCHER AND LESTER PACKER VOLUME LIV. Biomembranes (Part E: Biological Oxidations)

Edited by SIDNEY FLEISCHER AND LESTER PACKER VOLUME LV. Biomembranes (Part F: Bioenergetics)

Edited by SIDNEY FLEISCHER AND LESTER PACKER

lxxxvi

METHODS IN ENZYMOLOGY

VOLUME LVI. Biomembranes (Part G: Bioenergetics)

Edited by SIDNEY FLEISCHERAND LESTER PACKER VOLUME LVII. Bioluminescence and Chemiluminescence

Edited by MARLENE A. DELUCA VOLUME LVIII. Cell Culture

Edited by WILLIAM B. JAKOBYAND IRA PASTAN VOLUME LIX. Nucleic Acids and Protein Synthesis (Part G)

Edited by KIVIE MOLDAVE AND LAWRENCE GROSSMAN VOLUME LX. Nucleic Acids and Protein Synthesis (Part H)

Edited by KIVlE MOLDAVE AND LAWRENCE GROSSMAN VOLUME 61. Enzyme Structure (Part H)

Edited by C. H. W. HIRS AND SERGE N. TIMASHEFF VOLUME 62. Vitamins and Coenzymes (Part D)

Edited by DONALD B o McCORMICK AND LEMUEL D. WRIGHT VOLUME 63. Enzyme Kinetics and Mechanism (Part A: Initial Rate and Inhibitor Methods) Edited by DANIEL L. PURICH VOLUME 64. Enzyme Kinetics and Mechanism (Part B: Isotopic Probes and Complex Enzyme Systems)

Edited by DANIEL L. PURICH VOLUME 65. Nucleic Acids (Part I)

Edited by LAWRENCE GROSSMAN AND KIVIE MOLDAVE VOLUME 66. Vitamins and Coenzymes (Part E)

Edited by DONALD B. McCORMICK AND LEMUEL D. WRIGHT VOLUME 67. Vitamins and Coenzymes (Part F)

Edited by DONALD B. McCORMICK AND LEMUEL D. WRIGHT VOLUME 68. Recombinant D N A

Edited by RAY Wo VOLUME 69. Photosynthesis and Nitrogen Fixation (Part C)

Edited by ANTHONY SAN PIETRO VOLUME 70. Immunochemical Techniques (Part A)

Edited by HELEN VAN VUNAKIS AND JOHN J. LANGONE VOLUME 71. Lipids (Part C)

Edited by JOHN i . LOWENSTEIN VOLUME 72. Lipids (Part D)

Edited by JOHN m. LOWENSTEIN VOLUME 73. Immunochemical Techniques (Part B)

Edited by JOHN J. LANGONE AND HELEN VAN VUNAKIS

M E T H O D S IN E N Z Y M O L O G Y

lxxxvii

VOLUME 74. Immunochemical Techniques (Part C) Edited by JOHN J. LANGONEAND HELEN VAN VUNAKIS VOLUME 75. Cumulative Subject Index Volumes XXXI, XXXII, XXXIV-LX Edited by EDWARD A. DENNIS AND MARTHAG. DENNIS VOLUME 76. Hemoglobins

Edited by ERALDOANTONINI, LUIGI ROSSI-BERNARDI,AND EMIL1ACHIANCONE VOLUME 77. Detoxication and Drug Metabolism

Edited by WILLIAMB. JAKOBY VOLUME 78. Interferons (Part A)

Edited by SIDNEY PESTKA VOLUME 79. Interferons (Part B)

Edited by SIDNEY PESTKA VOLUME 80. Proteolytic Enzymes (Part C) Edited by LASZLOLORAND VOLUME 81. Biomembranes (Part H: Visual Pigments and Purple Membranes, I) Edited by LESTER PACKER VOLUME 82. Structural and Contractile Proteins (Part A: Extracellular Matrix)

Edited by LEON W. CUNNINGHAMAND DIXIE W. FREDERIKSEN VOLUME 83. Complex Carbohydrates (Part D)

Edited by VICTORGINSBURG VOLUME 84. Immunochemical Techniques (Part D: Selected Immunoassays)

Edited by JOHN J. LANGONEAND HELEN VAN VUNAKIS VOLUME 85. Structural and Contractile Proteins (Part B: The Contractile Apparatus and the Cytoskeleton) Edited by DIXIE W. FREDERIKSENAND LEON W. CUNNINGHAM VOLUME 86. Prostaglandins and Arachidonate Metabolites Edited by WILLIAME. M. LANDS AND WILLIAML. SMITH VOLUME 87. Enzyme Kinetics and Mechanism (Part C: Intermediates, Stereochemistry, and Rate Studies) Edited by DANIEL L. PURICH VOLUME 88. Biomembranes (Part I: Visual Pigments and Purple Membranes, II) Edited by LESTER PACKER VOLUME 89. Carbohydrate Metabolism (Part D) Edited by WILLIS A. WOOD VOLUME 90. Carbohydrate Metabolism (Part E) Edited by WILLISA. WOOD

lxxxviii

M E T H O D S IN E N Z Y M O L O G Y

VOLUME 91. Enzyme Structure (Part I)

Edited by C. H. W. HIRS AND SERGE N. TIMASHEFF VOLUME 92. Immunochemical Techniques (Part E: Monoclonal Antibodies and General Immunoassay Methods) Edited by JOHN J. LANGONEAND HELEN VAN VUNAKIS VOLUME 93. Immunochemical Techniques (Part F: Conventional Antibodies, Fc Receptors, and Cytotoxicity) Edited by JOHN J. LANGONEAND HELEN VAN VUNAKIS VOLUME 94. Polyamines

Edited by HERBERTTABORAND CELIA WHITE TABOR VOLUME 95. Cumulative Subject Index Volumes 61-74, 76-80

Edited by EDWARD A. DENNIS AND MARTHAG. DENNIS VOLUME 96. Biomembranes [Part J: Membrane Biogenesis: Assembly and Targeting (General Methods; Eukaryotes)] Edited by SIDNEY FLEISCHERAND BECCA FLEISCHER VOLUME 97. Biomembranes [Part K: Membrane Biogenesis: Assembly and Targeting (Prokaryotes, Mitochondria, and Chloroplasts)] Edited by SIDNEY FLEISCHERAND BECCA FLEISCHER VOLUME 98. Biomembranes (Part L: Membrane Biogenesis: Processing and Recycling) Edited by SIDNEY FLEISCHERAND BECCA FLEISCHER VOLUME 99. Hormone Action (Part F: Protein Kinases)

Edited by JACKIE D. CORBIN AND JOEL G. HARDMAN VOLUME 100. Recombinant DNA (Part B)

Edited by RAY Wu, LAWRENCEGROSSMAN, AND KIVIE MOLDAVE VOLUME 101. Recombinant DNA (Part C)

Edited by RAY Wu, LAWRENCEGROSSMAN,AND KIVIE MOLDAVE VOLUME 102. Hormone Action (Part G: Calmodulin and Calcium-Binding Proteins) Edited by ANTHONYR. MEANS AND BERT W. O'MALLEY VOLUME 103. Hormone Action (Part H: Neuroendocrine Peptides)

Edited by P. MICHAELCONN VOLUME 104. Enzyme Purification and Related Techniques (Part C)

Edited by WILLIAMB. JAKOBY VOLUME 105. Oxygen Radicals in Biological Systems

Edited by LESTER PACKER VOLUME 106. Posttranslational Modifications (Part A)

Edited by FINN WOLD AND KIVIE MOLDAVE VOLUME 107. Posttranslational Modifications (Part B)

Edited by FINN WOLD AND KIVIE MOLDAVE

METHODS IN ENZYMOLOGY

lxxxix

VOLUME 108. Immunochemical Techniques (Part G: Separation and Characterization of Lymphoid Cells) Edited by GIOVANNIDI SABATO,JOHN J. LANGONE, AND HELEN VAN VUNAKIS

VOLUME 109. Hormone Action (Part I: Peptide Hormones)

Edited by LUTZ BIRNBAUMERAND BERT W. O'MALLEY VOLUME 110. Steroids and Isoprenoids (Part A)

Edited by JOHN H. LAW AND HANS C. RILLING VOLUME 111. Steroids and Isoprenoids (Part B) Edited by JOHN H. LAW AND HANS C. RILLING VOLUME 112. Drug and Enzyme Targeting (Part A)

Edited by KENNETH J. WIDDER AND RALPH GREEN VOLUME ll3. Glutamate, Glutamine, Glutathione, and Related Compounds

Edited by ALTON MEISTER VOLUME 114. Diffraction Methods for Biological Macromolecules (Part A)

Edited by HAROLD W. WYCKOFF, C. H. W. HIRS, AND SERGE N. TIMASHEEF VOLUME 115. Diffraction Methods for Biological Macromolecules (Part B)

Edited by HAROLD W. WYCKOFF, C. H. W. HIRS, AND SERGE N. TIMASHEFF VOLUME 116. Immunochemical Techniques (Part H: Effectors and Mediators of Lymphoid Cell Functions) Edited by GIOVANNIDI SABATO, JOHN J. LANGONE, AND HELEN VAN VUNAKIS

VOLUME 117. Enzyme Structure (Part J)

Edited by C. H. W. HIRS AND SERGE N. TIMASHEFF VOLUME 118. Plant Molecular Biology

Edited by ARTHURWEISSBACHAND HERBERT WEISSBACH VOLUME 119. Interferons (Part C)

Edited by SIDNEY PESTKA VOLUME 120. Cumulative Subject Index Volumes 81-94, 96-101 VOLUME 121. Immunochemical Techniques (Part I: Hybridoma Technology and Monoclonal Antibodies) Edited by JOHN J. LANGONEAND HELEN VAN VUNAKIS VOLUME 122. Vitamins and Coenzymes (Part G)

Edited by FRANK CHYTIL AND DONALD B. McCoRMICK VOLUME 123. Vitamins and Coenzymes (Part H)

Edited by FRANK CHYTIL AND DONALD B, McCoRMICK VOLUME 124. Hormone Action (Part J: Neuroendocrine Peptides)

Edited by P. MICHAELCONN

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METHODS IN ENZYMOLOGY

VOLUME 125. Biomembranes (Part M: Transport in Bacteria, Mitochondria, and Chloroplasts: General Approaches and Transport Systems) Edited by SIDNEY FLEISCHERAND BECCA FLEISCHER VOLUME 126. Biomembranes (Part N: Transport in Bacteria, Mitochondria, and Chloroplasts: Protonmotive Force) Edited by SIDNEY FLEISCHERAND BECCA FLEISCHER VOLUME 127. Biomembranes (Part O: Protons and Water: Structure and Translocation) Edited by LESTERPACKER VOLUME 128. Plasma Lipoproteins (Part A: Preparation, Structure, and Molecular Biology) Edited by JERE P. SEGRESTAND JOHN J. ALBERS VOLUME 129. Plasma Lipoproteins (Part B: Characterization, Cell Biology, and Metabolism) Edited by JOHN J. ALBERSAND JERE P. SEGREST VOLUME 130. Enzyme Structure (Part K)

Edited by C. H. W. HIRS AND SERGE N. TIMASHEFF VOLUME 131. Enzyme Structure (Part L)

Edited by C. H. W. HIRS AND SERGE N. TIMASHEFF VOLUME 132. Immunochemical Techniques (Part J: Phagocytosis and CellMediated Cytotoxicity) Edited by GIOVANNIDI SABATO AND JOHANNES EVERSE VOLUME 133. Bioluminescence and Chemiluminescence (Part B)

Edited by MARLENEDELUCA AND WILLIAMD. MCELROY VOLUME 134. Structural and Contractile Proteins (Part C: The Contractile Apparatus and the Cytoskeleton)

Edited by

RICHARD B. VALLEE

VOLUME 135. Immobilized Enzymes and Cells (Part B)

Edited by KLAUS MOSBACH VOLUME 136. Immobilized Enzymes and Cells (Part C) Edited by KLAUS MOSBACH VOLUME 137. Immobilized Enzymes and Cells (Part D)

Edited by KLAUS MOSBACH VOLUME 138. Complex Carbohydrates (Part E)

Edited by VICTOR GINSBURG VOLUME 139. Cellular Regulators (Part A: Calcium- and Calmodulin-Binding Proteins) Edited by ANTHONY R. MEANS AND P. MICHAELCONN VOLUME 140. Cumulative Subject Index Volumes 102-119, 121-134 VOLUME 141. Cellular Regulators (Part B: Calcium and Lipids)

Edited by P.

MICHAEL CONN AND ANTHONY R. MEANS

METHODS IN ENZYMOLOGY

xci

VOLUME 142. Metabolism of Aromatic Amino Acids and Amines

Edited by SEYMOURKAUFMAN VOLUME 143. Sulfur and Sulfur Amino Acids

Edited by WILLIAMB. JAKOBYAND OWEN GRIFFITH VOLUME 144. Structural and Contractile Proteins (Part D: Extracellular Matrix)

Edited by LEON W. CUNNINGHAM VOLUME 145. Structural and Contractile Proteins (Part E: ExtraceUular Matrix)

Edited by LEON W. CUNNINGHAM VOLUME 146. Peptide Growth Factors (Part A)

Edited by DAVID BARNESAND DAVID A. SIRBASKU VOLUME 147. Peptide Growth Factors (Part B)

Edited by DAVID BARNESAND DAVID A. SIRBASKU VOLUME 148. Plant Cell Membranes

Edited by LESTER PACKER AND ROLAND DOUCE VOLUME 149. Drug and Enzyme Targeting (Part B)

Edited by RALPH GREEN AND KENNETH J. WIDDER VOLUME 150. Immunochemical Techniques (Part K: In Vitro Models of B and T Cell Functions and Lymphoid Cell Receptors) Edited by GIOVANNIDI SABATO VOLUME 151. Molecular Genetics of Mammalian Cells

Edited by MICHAEL M. GOTTESMAN VOLUME 152. Guide to Molecular Cloning Techniques

Edited by SHELBY L. BERGER AND ALAN R. KIMMEL VOLUME 153. Recombinant DNA (Part D)

Edited by RAY Wu AND LAWRENCEGROSSMAN VOLUME 154. Recombinant DNA (Part E)

Edited by RAY WU AND LAWRENCE GROSSMAN VOLUME 155. Recombinant DNA (Part F)

Edited by RAY Wu VOLUME 156. Biomembranes (Part P: ATP-Driven Pumps and Related Transport: The Na,K-Pump) Edited by SIDNEY FLEISCHER AND BECCA FEEISCHER VOLUME 157. Biomembranes (Part Q: ATP-Driven Pumps and Related Transport: Calcium, Proton, and Potassium Pumps)

Edited by SIDNEY FLEISCHER AND BECCA FLEISCHER VOLUME 158. Metalloproteins (Part A)

Edited by JAMES F. RIORDAN AND BERT L. VALLEE VOLUME 159. Initiation and Termination of Cyclic Nucleotide Action

Edited by JACKIE D. CORBIN AND ROGER A. JOHNSON

xcii

METHODS IN ENZYMOLOGY

VOLUME 160. Biomass (Part A: Cellulose and Hemicellulose)

Edited by WILLISA. WOOD AND SCOTT T. KELLOGG VOLUME 161. Biomass (Part B: Lignin, Pectin, and Chitin)

Edited by WILLISA. WOOD AND SCOTT T. KELLOGG VOLUME 162. Immunochemical Techniques (Part L: Chemotaxis and Inflammation) Edited by GIOVANNIDI SABATO VOLUME 163. Immunochemical Techniques (Part M: Chemotaxis and Inflammation) Edited by GIOVANNIDI SABATO VOLUME 164. Ribosomes

Edited by HARRY F. NOLLER, JR., AND KIVIE MOLDAVE VOLUME 165. Microbial Toxins: Tools for Enzymology

Edited by SIDNEY HARSHMAN VOLUME 166. Branched-Chain Amino Acids

Edited by ROBERT HARRIS AND JOHN R. SOKATCH VOLUME 167. Cyanobacteria

Edited by LESTER PACKERAND ALEXANDERN. GLAZER VOLUME 168. Hormone Action (Part K: Neuroendocrine Peptides)

Edited by P. MICHAELCONN VOLUME 169. Platelets: Receptors, Adhesion, Secretion (Part A)

Edited by JACEK HAWIGER VOLUME 170. Nucleosomes

Edited by PAUL M. WASSARMANAND ROGER D. KORNBERG VOLUME 171. Biomembranes (Part R: Transport Theory: Cells and Model Membranes) Edited by SIDNEY FLEISCHERAND BECCA FLEISCHER VOLUME 172. Biomembranes (Part S: Transport: Membrane Isolation and Characterization) Edited by SIDNEY FLEISCHERAND BECCA FLEISCHER VOLUME 173. Biomembranes [Part T: Cellular and Subcellular Transport: Eukaryotic (Nonepithelial) Cells] Edited by SIDNEY FLEISCHERAND BECCA FLEISCHER VOLUME 174. Biomembranes [Part U: Cellular and Subcellular Transport: Eukaryotic (Nonepithelial) Cells] Edited by SIDNEY FLEISCHERAND BECCA FLEISCHER VOLUME 175. Cumulative Subject Index Volumes 135-139, 141-167 VOLUME 176. Nuclear Magnetic Resonance (Part A: Spectral Techniques and Dynamics) Edited by NORMANJ. OPPENHEIMERAND THOMAS L. JAMES

. ° .

METHODS IN ENZYMOLOGY

XCII1

VOLUME 177. Nuclear Magnetic Resonance (Part B: Structure and Mechanism) Edited by NORMAN J. OPPENHEIMER AND THOMAS L. JAMES VOLUME 178. Antibodies, Antigens, and Molecular Mimicry

Edited by JOHN J. LANGONE VOLUME 179. Complex Carbohydrates (Part F) Edited by VICTOR GINSBURG VOLUME 180. RNA Processing (Part A: General Methods)

Edited by JAMES E. DAHLBERG AND JOHN N. ABELSON VOLUME 181. RNA Processing (Part B: Specific Methods)

Edited by JAMES E. DAHLBERG AND JOHN N. ABELSON VOLUME 182. Guide to Protein Purification

Edited by MURRAY P. DEUTSCHER VOLUME 183. Molecular Evolution: Computer Analysis of Protein and Nucleic Acid Sequences Edited by RUSSELL F. DOOLITTLE VOLUME 184. Avidin-Biotin Technology

Edited by MEIR WILCHEK AND EDWARD A. BAYER VOLUME 185. Gene Expression Technology

Edited by DAVID V. GOEDDEL VOLUME 186. Oxygen Radicals in Biological Systems (Part B: Oxygen Radicals and Antioxidants) Edited by LESTER PACKER AND ALEXANDER N. GLAZER VOLUME 187. Arachidonate Related Lipid Mediators

Edited by ROBERT C. MURPHY AND FRANK A. FITZPATRICK VOLUME 188. Hydrocarbons and Methylotrophy

Edited by MARY E. LIDSTROM VOLUME 189. Retinoids (Part A: Molecular and Metabolic Aspects)

Edited by LESTER PACKER VOLUME 190. Retinoids (Part B: Cell Differentiation and Clinical Applications)

Edited by LESTER PACKER VOLUME 191. Biomembranes (Part V: Cellular and Subcellular Transport: Epithelial Cells) Edited by SIDNEY FLEISCHER AND BECCA FLEISCHER VOLUME 192. Biomembranes (Part W: Cellular and Subcellular Transport: Epithelial Cells) Edited by SIDNEY FLEISCHER AND BECCA FLEISCHER VOLUME 193. Mass Spectrometry

Edited by JAMES A. McCLOSKEY VOLUME 194. Guide to Yeast Genetics and Molecular Biology

Edited by CHRISTINE GUTHRIE AND GERALD R. FINK

xciv

METHODS IN ENZYMOLOGY

VOLUME 195. Adenylyl Cyclase, G Proteins, and Guanylyl Cyclase

Edited by ROGER A. JOHNSONAND JACKIE D. CORBIN VOLUME 196. Molecular Motors and the Cytoskeleton

Edited by RICHARDB. VALLEE VOLUME 197. Phospholipases

Edited by EDWARD A. DENNIS VOLUME 198. Peptide Growth Factors (Part C)

Edited by DAVID BARNES, J. P. MATHER, AND GORDON H. SATO VOLUME 199. Cumulative Subject Index Volumes 168-174, 176-194 VOLUME 200. Protein Phosphorylation (Part A: Protein Kinases: Assays, Purification, Antibodies, Functional Analysis, Cloning, and Expression) Edited by TONY HUNTER AND BARTHOLOMEWi . SEFTON VOLUME 201. Protein Phosphorylation (Part B: Analysis of Protein Phosphorylation, Protein Kinase Inhibitors, and Protein Phosphatases) Edited by TONY HUNTER AND BARTHOLOMEWi . SEFTON VOLUME 202. Molecular Design and Modeling: Concepts and Applications (Part A: Proteins, Peptides, and Enzymes) Edited by JOHN J. LANGONE VOLUME 203. Molecular Design and Modeling: Concepts and Applications (Part B: Antibodies and Antigens, Nucleic Acids, Polysaccharides, and Drugs) Edited by JOHN J. LANGONE VOLUME 204. Bacterial Genetic Systems

Edited by JEFFREY H. MILLER VOLUME 205. Metallobiochemistry (Part B: Metallothionein and Related Molecules) Edited by JAMES F. RIORDANAND BERT L. VALLEE VOLUME 206. Cytochrome P450

Edited by MICHAELR. WATERMANAND ERIC F. JOHNSON VOLUME 207. Ion Channels

Edited by BERNARDORUDY AND LINDA E. IVERSON VOLUME 208. Protein-DNA Interactions

Edited by ROBERT T. SAUER VOLUME 209. Phospholipid Biosynthesis

Edited by EDWARD A. DENNIS AND DENNIS E. VANCE VOLUME 210. Numerical Computer Methods

Edited by LUDWIG BRAND AND MICHAELL. JOHNSON VOLUME 211. DNA Structures (Part A: Synthesis and Physical Analysis of DNA) Edited by DAVID M. J. LILLEY AND JAMES E. DAHLBERG

METHODS IN ENZYMOLOGY

XCV

VOLUME 212. DNA Structures (Part B: Chemical and Electrophoretic Analysis of DNA)

Edited by

DAVID M. J. LILLEY AND JAMES E. DAHLBERG

VOLUME 213. Carotenoids (Part A: Chemistry, Separation, Quantitation, and Antioxidation) Edited by LESTER PACKER VOLUME 214. Carotenoids (Part B: Metabolism, Genetics, and Biosynthesis) (in preparation) Edited by LESTER PACKER VOLUME 215. Platelets: Receptors, Adhesion, Secretion (Part B)

Edited by JACEK HAWIGER VOLUME 216. Recombinant DNA (Part G)

Edited by RAY Wu VOLUME 217. Recombinant DNA (Part H)

Edited by RAY Wu VOLUME 218. Recombinant DNA (Part I) (in preparation)

Edited by RAY Wu VOLUME 219. Reconstitution of Intracellular Transport

Edited by JAMES E. ROTHMAN VOLUME 220. Membrane Fusion Techniques (Part A) (in preparation)

Edited by NEJAT DOZG~NES VOLUME 221. Membrane Fusion Techniques (Part B) (in preparation)

Edited by NEJAT D~ZGt3NES

Subject Index Boldface numerals indicate volume number.

A A23187 behavior and effects in red cells, 173, 103 induced Ca 2+ permeabilization of ATP-depleted red cells, 173, 110 red cells, 173, 107 Absorptiometry malondialdehyde, 186, 423 Absorption salt by teleost intestine, model, 192, 752 turtle colon, model, 192, 744 Absorption spectroscopy amiloride analogs, 191, 749 choleglobin, 186, 270 ferrylhemoglobin, 186, 270 free radicals, 186, 92 bemichrome, 186, 270 hydroxyl radicals, 186, 143 methemoglobin, 186, 268 oxyhemoglobin, 186, 268 Abzymes, see Antibodies, catalytic Acetabularia

ATP-driven chloride pump, analysis, 174, 490 culture and handling, 174, 490 Acetate buffer stock solutions, 182, 32 -glycine buffers, nucleotide peptides in, gel electrophoresis, 170, 407 Acetic acid -Triton X-100-urea gels, preparation, 170, 544 -urea gels, preparation, 170, 535 Acetobacter methanolicus MB58 purification of 3-hexulose-6-phosphate synthase, 188, 402 Acetol dehydrogenase assay, 188, 31 Acetol monooxygenase assay, 188, 30

Acetolysis mannoproteins, 185, 467 peracetylated high mannose structures. 193, 653 Acetone and acid, precipitation of proteins, 182, 592 powder, preparation, 174, 55 Acetone monooxygenase assay, 188, 30 9-O-Acetyl-N-acetylneuraminic acid ~H-labeled at acetyl groups in assay of sialate 9-O-acetylesterase. 179, 409 preparation, 179, 409 Acetylation alditol acetates, 193, 560 for mass spectrometry, 193, 326, 328 peptide fragments, 193, 478 proteins, inhibition, 185,406 in structural analysis of porcine mucin, 179, 270 Acetylcholine receptors autoimmune reactivity, inhibition with idiotype-specific immunotoxins. 178, 448 nicotinic anti-idiotypic antibodies, 178, 24 characterization with amiloride. 191. 746 oligosaccharides from, MS analysis. 193. 679 Acetylcholinesterase in immunoassay of eicosanoids, 187, 24 leukotrienes C4 and E4, 187, 82 Acetylcholinesterase receptors in myasthenia gravis anti-idiotypes detection and characterization, 178. 437 preparation, 178, 434 modulation by anti-idiotypes, 178, 444

N-Acetylgalactosamine N-Acetylgalactosamine -galactose-binding protein filter assay, 179, 323 isolation from rat peritoneal macrophages, 179, 324 Acetylgalactosaminyl-O-glycosyl-glycoprotein fl- 1,3-N-acetylglucosaminyltransferase assay, 179, 386 properties, 179, 388 Acet ylgalactosaminyl-O-glycosyl-glycoprotein fl- 1,6-N-acetylglucosaminyltransferase assay, 179, 389 rat colon, properties, 179, 390 N-Acetylglucosamine -mannose-binding proteins, purification from mammalian sera, 179, 310 -mannose/L-fucose-binding protein filter assay, 179, 323 isolation from rat peritoneal macrophages, 179, 324 O-linked, in proteins, analysis with glycosyltransferases, 179, 93 pyridylaminated, in assay of glycosyltransferases, 179, 401 N-Acetyl-fl-glucosaminidase lysosomal marker, assay, 182, 215 secretion by platelets assay, 169, 436 effects of diacylglycerol and Ca 2+ ionophore, 169, 436 N-Acetylglucosaminyltransferase II, s e e a- 1,6-Mannosyl-glycoprotein fl-l,2-Nacetylglucosaminyltransferase N-Acetylglucosaminyltransferase III, s e e fl- 1,4-Mannosyl-glycoprotein fl- 1,4-Nacetylglucosaminyltransferase N-Acetylglucosaminyltransferase IV, s e e a-1,3-Mannosyl-glycoprotein fl-1,4-Nacetylglucosaminyltransferase N-Acetylglucosaminyltransferase V, s e e a- 1,6-Mannosyl-glycoprotein fl- 1,6-Nacet ylglucosaminyltransferase N-Acetylglucosaminyltransferases in N- and O-glycan synthesis HPLC assay, 179, 351 substrate preparation, 179, 360 fl-N-Acetylhexosaminidase secretion from platelet lysosomes, assay, 169, 336

2 N-Acetyllactosaminide fl- 1,3-N-acetylglucosaminyltransferase assay, 179, 393 blood group i partial purification, 179, 394 properties, 179, 394 N-Acet yl-DL-phenylalanine fl-naphthyl ester based overlay test for yeast protease B activity, 194, 437 N-Acetyl-L-valyl-L-leucine single crystal, backbone structure, solidstate NMR study, 176, 258 Acid glycosidases secretion from platelet lysosomes, assay, 169, 336 Acid hydrolases secretion by platelets, assay, 169, 196 Acidity free radicals, 186, 92 Acid phosphatase -streptavidin, in detection of antigens on Western blots, 1114, 441 Acids, s e e also s p e c i f i c a c i d s Brensted, reagents for chemical ionization, 193, 14 catalyzed ~sO exchange of amino acids, 193, 341 keto groups, 193, 343 phenols, 193, 346 loading for fluorimetric assay of Na+/H + exchange, 173, 787 mechanisms, identification in parietal cells, 192, 55 organic, in yeast suspension filtrates, estimation, 174, 602 production by parietal cells, effects of prostaglandins, 187, 512 secretion by permeabilized parietal cells, 192, 112, 121 -urea gels, in first-dimension separation of RNA fingerprinting, 180, 143 weak, and weak bases, equilibrium distribution, 172, 79 Acinar cells leakiness, assessment, 192, 290 pancreatic atrophy, 192,259 dispersed applications, 192, 273

3

Adenylate cyclase preparation, 192, 271 electrophysiology, 192, 300 fluid secretion, role of ionic currents, 192, 306 intracellular Ca 2+ levels oscillations, analysis, 192, 304 stimulant-evoked increase, 192, 301 isolation, 192, 288 permeabilization by digitonin or saponin, 192, 283, 292 high-voltage electric discharge, 192, 289 streptolysin O and a-toxin, 192, 285, 296 salivary gland neuroreceptor-coupled secretory events, 192, 26 preparation, 192, 26 protein secretion, assay, 192, 31 water transport, related ion fluxes, assay, 192, 33

Acinetobacter

purification of catechol 1,2-dioxygenase, 188, 124 HO1-N long-chain alkane oxidation, assay, 188, 10 preparation for assay of aldehyde dehydrogenase, 188, 19 primary alcohol dehydrogenases, 188, 15 NCIMB 9871, purification of cyclohexanone 1,2-monooxygenase, 188, 72 Acini pancreatic, as second messenger models in exocrine secretion, 171, 590 Acitretin pharmacokinetics, 190, 293 Acquired immune deficiency syndrome, s e e Human immunodeficiency virus Acridine orange in measurement of pH gradients in cells and vesicles, 191,474 dissipation, 192, 403 Acridines fluorescence, in determination of biomembrane surface potential, 171, 367 A. calcoaceticus,

Acrylylation glycoside, in carbohydrate immobilization on polyacrylamide gels, 179, 544 ACTH, s e e Adrenocorticotropin Actin staining with fluorochrome-conjugated phalloidin, 194, 729 Action potentials in giant algal cells, analysis, 174, 413 Acylation protein-bound fatty acids, 185, 593 protein by retinoic acid, 189, 233 Acylazides generated nitrenes, properties, 172, 638 Acyl-CoA:retinol acyltransferase, s e e Retinol fatty-acyltransferase Acyl coenzyme A dependent retinol esterification, assay, 189, 442,446 Acylglycerol lipase role in arachidonate liberation from phospholipids, 191, 677 Adenine in RNA, modification by diethyl pyrocarbonate, 180, 374 Adenine nucleotides ATP/3,yNH, caged, synthesis and properties, 172, 289 in CaZ+-tolerant atrial and ventricular myocytes, assay, 173, 673 levels in C h a r a cells, effect on ion transport, 174, 472 secretion by platelets, assay, 169, 195 Adenosine 3',5'-cyclic monophosphate, s e e Cyclic AMP Adenosine deaminase binding protein, competitive idiotypeanti-idiotype enzyme immunoassay, 178, 171 in heterologous gene selection and coamplification in mammalian cells, 185, 552 Adenosine receptors probing with biotinylated purine derivatives, 184, 668 Adenosinetriphosphatase, s e e ATPase Adenylate cyclase activation in gastric parietal cells. 191, 649

Adenylate cyclase characterization with amiloride, 191, 746 vasopressin-sensitive, target analysis studies, 191,566 Adenylate kinase bound reactants and products, equilibrium constant determination, 177, 365 31p exchanges in, 176, 303 substrate binding, quadrupolar NMR study, 177, 331 Adenylylation glutamine synthetase, 182, 795 Adenylyltransferase identity with Holzer's inactivation enzyme, 182, 798 role in adenylylation and deadenylylation of glutamine synthetase, 182, 797 Adipocytes demonstration of fructose transporter, 173,629 hexose transport, assay, 173, 623 purification of lipid-binding proteins from human and mouse, 189, 363 ADP (adenosine 5'-diphosphate) -ATP exchange in mitochondria, characterization, 171, 12 effect on citrated platelet-rich plasma, 169, 131 induced preformed platelet aggregates, adherence to endothelial cell monolayers, 169, 75 photoreactive biotinylated analog synthesis, 184, 409 in visualization of myosin ATPase site of myosin, 184, 409 ADP-ribosylation effective diphtheria toxin A chain linked to monoclonal antibody, assay, 178, 414 proteins, overview, 182, 637 Adrenal medulla detection of preproenkephalin and tyrosine monooxygenase transcripts, 168, 688 ~-Adrenergic receptors characterization with amiloride and analogs, 191, 746 fl-Adrenergic receptors anti-idiotypic antibodies, 178, 25

4 characterization with amiloride and analogs, 191, 746 probes, preparation and characterization, 184, 660 Adrenocorticotropin binding proteins assay, 184, 267 biotinylation, 184, 264 isolation, 184, 262 properties, 184, 270 Adrenocorticotropin receptors antibodies, production with complementary peptides, 168, 25 Adriamycin, s e e Doxorubicin Adsorption glycosphingolipids on plastic microwells, 179, 546 immuno-, s e e Immunoadsorption isocratic, in HPLC of retinyl esters and geometric isomers, 189, 181 plasma desorption MS sample on nitrocellulose, 193, 434 proteins on gold particles, 179, 114 hydroxyapatite, 182, 330 SV40 minichromosomes or nucleosomes on electron microscope grid, 170, 173 Adsorption chromatography histone complexes, 170, 442 RNA on hydroxylapatite, 180, 29 A e d e s a e g y p t i , s e e Mosquito Aequorin in assay of platelet [Ca2+]~, 169, 387 as intracellular Ca2+ indicator, 172, 165 introduction into cells, 169, 389 platelet integrity during and after loading with, 169, 391 preparation and handling, 169, 389 and Quin 2, signal comparison, 169, 409 Affinity chromatography ACTH-binding proteins, 184, 269 angiotensin II, 168, 550 angiotensin II antiserum, 168, 547 antibodies for immunofluorescence studies of yeast, 194, 584 avidin and biotinyl proteins, 184, 53 derivatives, 184, 171 nonglycosylated form, 184, 69

5

Agarose band 3 protein, 173, 505 with biotinylated RNAs, 181, 97 biotinyl proteins, 168, 53 cell isolation, overview, 182, 371 chloride channel, 191, 577 citrate synthase, 188, 351 covalent, overview, 182, 376 DNA-binding proteins, 182, 521 formaldehyde dehydrogenase with cofactor requirement, 188, 322 fusion proteins from Escherichia coli, 185, 136 B-galactosidase fusion proteins, 182, 110 glycolipids, 179, 30 glycoproteins from cell surface, indirect methods, 184, 305 lectin-based methods, 182, 530 gonadotropin-releasing hormone receptors, 184, 290 high-performance liquid, oligosaccharides, 179, 55 hormone receptors, 184, 636 idiotypes for analysis of anti-idiotype antibodies to hormone receptors, 178, 197 immobilized metal overview, 182,373 proteins, 182, 417 immunological probes for nucleosomes, 170, 226 insulin-like growth factor-lI receptors, 168, 314 insulin receptors, 184, 255 integral membrane proteins, 182, 510 with monovalent avidin columns, 184, 199 neurohypophyseal peptides, 168, 585 oligosaccarides on Allo A-II-agarose column, 179, 335 on Ea-phytohemagglutinin-agarose column, 179, 46 glycolipid-derived, 179, 30 high-performance methods, 179, 55 polyclonal anti-idiotypic anti-morphine antibodies, 178, 234 proteins column sterilization, 182, 369 concentration and solute removal, 182, 83

general methods, 182, 357 from hnRNP complexes, 181, 326 on immobilized reactive dyes, 182. 343 recombinant antifibrin antibody with t-PA activity, 178, 524 recombinant Fv and Fab fragments, 178. 510 replicating SV40 chromosomes, 170, 41 RNA, 180, 34 scale-up, 182, 377 site-specific antipeptide antibodies to rhinovirus receptor, 178, 681 snowdrop lectin, 179, 328 splicing complexes, 184, 321 streptavidin derivatives, 184, 171 native form, 184, 84 superoxide dismutase, 186, 255 toluene dioxygenase, 188.42 Z-DNA, 184, 328 Affinity columns agarose-E4-phytohemagglutinin, preparation, 179, 46 2-iminobiotin, preparation, 184, 169 indanyloxyacetic, synthesis. 191, 576 insulin, properties, 184, 258 Affinity cytochemistry overview, 184, 340 Affinity labeling catalytic antibodies, 178, 560 photo-, see Photoaffinity labeling Affinity labels photo-, see Photoaffinity labels for thiolation of catalytic antibodies, 178, 555 Affinity matrices amiloride-based, 191, 753 for chromatography of proteins, selection, 182, 358 African clawed frog care and maintenance, 170, 613 Agarose -Allomyrina dichotoma lectin type II affinity column, preparation, 179, 334 -avidin characteristics, 184, 196 preparation, 184, 195 synthesis, 184, 297

Agarose -E4-phytohemagglutin affinity column, preparation, 179, 46 Agarose gels conjugation of monoclonal anti-5-1ipoxygenase antibody to, 187, 341 formation and properties, 182, 462 preparation of chromosome-sized DNA in, 194, 60 threads, in NMR studies of intracellular metabolism, 177, 438 Agglutination -aggregation, platelets, tests with fixed platelets, 169, 149 assay with gold complexes, carbohydrate-specific binding, 179, 120 yeast mnn mutants, 185, 447 Agglutinins peanut, biotinylation, 184, 140 Aggregation number detergents, 182, 244 Agriculture practices, effects on retinol and provitamin A carotenoid content in food, 190, 237 AIDS, see Human immunodeficiency virus Air -liquid interface, keratinocyte culturing at, 190, 23 Alamethicins hybrid sector-quadrupole MS, 193, 193 Alanine formation in intact cells, in assay of protein degradation, 186, 497 spin system identification, 177, 135 transport across hepatocyte plasma membranes, 174, 31 role in regulation of Ala metabolism, 174, 33 Alanine aminotransferase nuclear labels, indirect detection with spin echoes, 176, 347 Albumin bound bilirubin, antioxidant activity, 186, 307 bound leukotriene A4, metabolism into leukotriene C4 by mast cells, 187, 575 gradients, with gel filtration, in isolation of human platelets, 169, 11 serum, see Serum albumin

6 Alcian blue in assay of yeast mnn mutants, 185, 450 coated carbon grids for chromatin electron microscopy, preparation and use, 170, 144 Alcohol dehydrogenase assay, 188, 29 cytosolic, differentiation from retinol dehydrogenase, 189, 478 isoenzymes in retinoid metabolism assay, 189, 437 purification from rat, 189, 438 starch gel electrophoresis, 189,438 long-chain assay, 188, 172 isolation from Candida, 188, 173 properties, 188, 174 NAD(P)+-dependent, soluble form assay, 188, 15 properties, 188, 16 propane-specific assay, 188, 21 properties, 188, 25 purification from Rhodococcus rhodochrous PNKbl, 188, 23 Alcohol oxidase assay, 188, 421 immunocytochemical detection, 188, 417 localization in Hansenula polymorpha, 188, 414 properties, 188, 424 purification from Hansenula polymorpha CBS 4732, 188, 422 Alcohol trapping in assay of leukotriene A4 release by neutrophils, 187, 572 Aldehyde dehydrogenase assay, 188, 29 Hyphornicrobium X, NAD-linked assay, 188, 328 properties, 188, 329 purification, 188, 329 long-chain assay, 188, 176 properties, 188, 177 NAD +- and NADP+-dependent assay, 188, 19 properties, 188, 20 Aldehyde dehydrogenase (acceptor) assay, 188, 323

7 properties, 188, 326 purification from Hyphomicrobium X, 188, 325 Aldehydes assay via dinitrophenylhydrazone formation, 186, 415 Alditol acetates partially methylated, identification, 193, 561 preparation, 193, 559 Aldolase, see Fructose-bisphosphate aldolase Aldosterone binding to isolated nephron segments, microassay, 191, 315 mineralocorticoid receptors, analysis, 191, 521 clearance studies, 191, 536 Algae, see also specific algae general characteristics, 174, 366 giant cells banding, 174, 427 cell compartments, control, 174, 433 cytoplasmic perfusion, 174, 437 electrical contact, 174, 403 membrane potential control, 174, 413 measurement, 174, 406 plasmalemma, permeabilization, 174, 440 vacuolar perfusion, 174, 434 transport systems, overview, 174, 366 Alizarin red S staining of rat fetal skeleton, 190, 420 Alkali metals -CI- cotransport, analysis, 173, 345 - H + exchange electroneutrality assessment, 173, 336 kinetics, 173, 337, 344 net flux studies on Amphiuma erythrocytes, 173, 331 pharmacological tests, 173, 336, 343 thermodynamics, 173, 340, 345 ion transport in Rhodotorula, 174, 645 NMR properties, 171, 317 Alkaline phosphatase biotinylation, 184, 149, 474

Amicyanin Alkaline phosphodiesterase plasma membrane marker, assay, 182, 218 Alkanes long-chain, oxidation in Acinetobacter, assay, 188, 10 Alkoxyl radicals kinetics and mechanisms, 186, 97 reactions in solution, rate constants, 186, 114 Alkoxy radicals role in cytochrome P450 peroxygenase activity, 186, 277 reductive cleavage of hydroperoxides by cytochrome P450. 186, 282 N-Alkyl p-aminobenzoates synthesis. 193, 664 Alkylation cysteine for amino acid analysis, 182. 598 proteins for mass spectrometry. 193. 373 Alkylazides properties. 172, 640 Alkylperoxyl radicals reactions in solution, rate constants, 186~ 114 Allergens Dermatophagoides pteronyssinus

antibodies immunoassay, 178, 155 preparation, 178, 153 preparation, 178, 151 Allium cepa, see Onion Allomyrina dichotoma, see Beetle Alprenolol antibodies anti-idiotypic, responses in mouse, 178, 272 rabbit, 178, 269 properties, 178, 266 idiotypic network components, 178, 274 Alum adsorbed immunoglobulins coupled to carrier protein, precipitation, 178. 46 Amicyanin assay, 188, 285 properties, 188, 288 purification from organism 4025, 188, 285

Amidation Amidation carboxy-terminal neuropeptides, FAB/MS study, 168, 79 proteins, analysis, 182, 628 Amido Black 10B in assay of proteins with high levels of lipids, 172, 393 Amiloride in characterization of enzymes, receptors, and cellular metabolism, 191, 746 ion transport systems, 191, 739 in identification of alkali metal/proton exchange, 173, 336, 343 inhibition of DNA, RNA, and protein synthesis, 191, 748 probe characteristics and types, 191,749 Aminacrine in measurement of pH gradients in cells and vesicles, 191,475 Amine oxidase (copper-containing) Arthrobacter P1 assay, 188, 227 properties, 188, 232 purification, 188, 229 Candida boidinii

assay, 188, 427 properties, 188, 433 purification, 188, 429 Amines cationic sugar, accumulation by Rhodotorula, energetics, 174, 643 3H-labeled, preparation, 191,765 Amino acid analyzers overview, 182, 589 Amino acid composition analysis, overview, 182, 587 catechol 2,3-dioxygenase, 188, 119 snowdrop lectin, 179, 328 Amino acids bacterial strains auxotrophic for, 177, 55 cellular, in protein degradation assay, 186, 498 contact residues, mimicry on anti-idiotypic antibodies, 178, 28 deletions and substitutions in monoclonal antibody modeling, restrictions, 178, 104 deuteration, 177, 34

8 -DNA base crosslinks, GC/MS analysis, 193, 850 isotopically labeled, incorporation into staphylococcal nuclease and sitedirected mutants, 177, 77 loading of lysosomes, 174, 156 metabolism, repression and inhibition, 177, 54 in mucin glycoproteins, analysis, 179, 14 -Na + symport in murine ascites tumor cells, 173, 771 t~N-labeled, availability, 177, 61 ~80 labeling by acid-catalyzed exchange with H2~80, 193, 341 oxidative modification, 186, 491 in peptides, mass values, 193, 888 phosphorylated, 31p NMR, 177, 268 physalaemin-related immunoreactive peptides, analysis, 168,454 in proteins, biosynthetic enrichment with 15N, 177, 54 -quinone adducts, o-semiquinone radicals from, characterization, 186, 339 spin systems, identification, 177, 133 total, in hydrolyzed samples, assay, 186, 470 Amino acid sequence alignment chained pairwise approach, 183, 457 computer analyses, 182, 752 consensus analysis, 183, 221 fast, algorithms for, 183, 487 flexible patterns, 183, 420 multiple sequences guidelines and accuracy assessment, 183,420 hierarchical method, 183, 456 sensitivity comparison, 183, 357 simultaneous methods, 183, 410 and phylogeny reconstruction, unified approach, 183, 626 by profile analysis, 183, 152 progressive, and phylogenetic tree construction, 183, 375 quality assessment, 183, 405 with secondary structure-dependent gap penalties, 183, 408 significance, 183, 361 three-way, with Needleman-Wunsch algorithm, 183, 367

9

Amino acid transport amino-terminal cytochrome P450 form 5, 187, 265 cytochrome P450 prostaglandin whydroxylase, 187, 265 protein purification for, 182, 602 requirements, 182, 603 strategies, 182, 603 a-amylases, saccharifying and liquefying, comparison, 183, 452 arachidonate 5-1ipoxygenase-derived peptides, 187, 492 biotinylated peptides, 184, 281 cell surface receptors, 191, 636 comparisons aggregate similarity scores, 183, 141 with FASTA and FASTP programs, 183, 63 general methods, 183, 404 pairwise, with Needleman-Wunsch algorithm, 183, 366 consensus based complementary peptide synthesis, 168, 19 definition, 183, 463 databases National Biomedical Research Foundation, 183, 31 searches, 183, 99 SWISS-PROT, 183, 26 hemoglobin, evolutionary tree construction, 183, 604 histones, 170, 503 homologous, aligned, evolutionary tree construction from, 183, 601 homology domains detection, 183, 136 reporting and display, 183, 139 in sequence similarity detection, 183, 133 interstitial retinol-binding protein bovine-derived tryptic peptides, 189, 212 human, 189, 208 multiple sequences alignment guidelines and accuracy assessment, 183, 420 methods, 183, 410 comparison methods, 183, 439 simultaneous comparison, 183, 447

patterns searches, 183, 193 statistically significant, identification, 183, 388 peptides derivation from FAB/MS fragmentation ions, 168, 101 STP-2, 193,407 tandem MS and high-energy collisioninduced dissociation methods. 193, 455 phosphopeptides and sulfopeptides, 193. 490 profile analysis, 183, 149 progressively aligned, analysis by nearest neighbor methods, 183, 659 prostaglandin endoperoxide synthases from mouse and sheep, 187. 477 proteins in prediction of B-cell determinants. 178, 586 tandem MS strategies, 193, 464 2D NMR methods, 176, 50 residue characteristic plots, 182, 772 sequence space, statistical geometry, 183, 505 similarity computer evaluation, 183.85 detection with homology domains, 183, 133 nucleotide sequence databases, 183. 111 local, identification, 183, 88 significance, 183,474 in superfamilies, profile analysis, 183, 158 superoxide dismutase, phylogeny determination, 183, 622 yeast ribosomal proteins, 194, 457 Amino acid transport by bacteria, 177.59 by Chlorella energization, 174, 396 inducibility and specificity, 174, 391 influx and efflux, assays, 174, 390, 392 by hepatocytes in suspension or monolayer culture, assay, 173, 564 pathway delineation, 172, 33 in red blood cells anion effects, 173. 146

Amino acid transport cell maturation effects, 173, 156 flux measurements, 173, 130 inhibitor studies, 173, 147 kinetics, 173, 138, 149 radioassay, 173, 133 role of ions, 173, 142 systems in, separation, 173, 125 thermodynamic effects, 173, 156 systems for abbreviated designations, proposal, 173, 580 heterogeneity, recognition and resolution, 173, 576 nomenclature and classification, 173, 123 in yeast, 174, 623 9-Aminoacridine, see Aminacrine p-Aminoazobenzene induced H202 formation, assay in vitro, 186, 625 p-Aminobenzoylbiocytin preparation, 184, 132 p-Aminobenzylphloretin synthes|s, 191, 759 p-Aminobenzylphlorizin synthesis, 191, 760 3 '-(4-Aminobenzyl)-2',4' ,6' ,4-tetrahydroxydihydrochalcone, see p-Aminobenzylphloretin 6-[N-(4-Aminobutyl-N-ethyl)-2,3-dihydrophthalazine- 1,4-dione] hemisuccinamide ester preparation and conjugation to streptavidin, 184, 487 Aminoethyl-Sepharose synthesis, 173, 502 Amino groups peptide, acetylation, 193, 328 protein, spin labeling, 177, 108 Aminohexyl-Sepharose CL-4b preparation, 184, 82 2-Amino-2-methyl-1,3-propanediol buffer stock solutions, 182, 36 Aminopeptidases Saccharomyces cerevisiae

enzymatic assay, 194, 450 overlay test, 194, 442 4-Aminophlorizin synthesis, 191,763

l0 Aminopyrene ~4C-labled, accumulation by gastric parietal cells, assay, 191, 647 p-Aminosalicylic acid staining of glycoproteins, 182, 536 Aminotransferases in selective ~N labeling of proteins by residue type, 177, 58 Ammoniacal silver solution preparation, 194, 728 Ammonium sulfate fractionation of potato tuber supernatant, 187, 271 protein bulk precipitates, 182, 293 Amnion human, isolation of phospholipase C 86kDa isoform, 187, 227 AMP (adenosine 5'-monophosphate) -RNA ligase complex, assay and characterization, 181, 497 Amperometric detection in affinity chromatography of oligosaccharides, 179, 61 and high-performance ion-exchange chromatography glycoconjugate monosaccharides, 179, 76 oligosaccharides, 179, 65 Amphipathic helix hypothesis T-cell epitope structure, 178, 616 AMPHI program T-cell epitope identification, 178, 617, 628 Ampholytes carrier, in isoelectric focusing of proteins, 182, 464 separation from protein, 182, 389, 477 Amputation limbs, 190, 189 a-Amylase liquefying and saccharifying, multiple sequence comparisons, 183, 452 secretion by pancreas, stimulation by secretagogues, 192, 248 Anaphylaxis cardiac, model of eicosanoid release and effects, 187, 619

11 Anesthesia rabbit and rat, 192, 4 for in vivo electrophysiological analysis of opioid peptides, 168, 104 Anesthetics effects on ex vivo studies of platelet adhesion to polymeric surfaces, 169, 101 Angiogenesis in skin. effect of retinoic acid, 190, 381 Angiotensin cerebral, chromatographic characterization, 168, 545 metabolites, reversed-phase HPLC, 168, 563 -renin system in juxtaglomerular interstitial cells, assay, 191, 156 tyrosine residues, phosphorylation by pp60 Src, 193, 485 Angiotensin 1 antibodies, cross-reacting material, reversed-phase HPLC, 168, 559 in cultured rat brain cells biosynthesis, 168, 555 degradation, 168, 561 Angiotensin I1 antibodies cross-reacting material, reversedphase HPLC, 168, 559 purification, 168, 546 antiserum, characterization, 168, 549 in cultured rat brain cells biosynthesis, 168, 555 degradation, 168, 561 isolation and purification, 168, 550 Angiotensin 1II degradation in cultured rat brain cells, 168, 561 Angiotensin II receptors anti-idiotypic antibodies to, 178, 25 Angiotensins dose-response testing by hippocampal brain slice recording, 168, 138 Anhydrides mixed, in preparation of leukotriene B4 immunogens, 187, 121 leukotriene D4 immunogens, 187, 120

Anions 1,5-Anhydro-D-glucitol partially methylated, GC/MS analysis. 193, 576

1,5-Anhydro-D-mannitol partially methylated. GC/MS analysis. 193, 576 Aniline cleavage of RNA modified by diethyl pyrocarbonate or hydrazine. 180. 375

l-Anilino-8-naphthalenesulfonate fluorescence, in determination of biomembrane surface potential, 171. 370 optical probe for membrane potential determination, 172, 76 2-Anilinonaphthalene-6-sulfonic acid -avidin complex titration by biotin, 184, 235 titration of biotin, 184. 239 Animal cells partitioning in aqueous polymer twophase systems. 171, 543 Animal models for peptide regulation of autonomic nervous system, 168, 434 Anions in buffers for cell permeabilization studies, selection, 192. 297 and cations, cotransport electrogenicity and electroneutrality. 173, 287 stoichiometry, 173, 283 lipophilic, in determination of yeast membrane potential, 174, 616 mass spectrometric detection, 193, 74 organic, transport across reptilian proximal tubules, mechanism, 191. 214 transport analytical methodology, 173. 401 in erythrocytes analysis, 173, 54 and ghosts, measurement, 173, 160 steady-state rate equations, generation, 171, 177 erythroid band 3-mediated, measurement in mRNA-injected Xenopus oocytes, 173. 453 in yeast, 174. 623

Anion transporter Anion transporter interaction with p-azidobenzylphloretin and p-azidobenzylphlorizin, 191, 776 modification with phenylglyoxal, 173, 466 for organic anions, isolation from hepatic plasma membrane, 174, 50 proteolytic cleavage and orientation in membrane, 173, 423 Annealing oligonucleotide primers to template, 180, 128 pre-mRNA and anchor RNA, 181, 106 ANP, s e e Atrial natriuretic factor Anterior pituitary CRF-target cells flow microfluorimetry, 168, 34 fluorescence microscopy, 168, 33 heterogeneous populations, probes, 168, 29 labeling, 168, 31 sorting, 168, 37 neuroendocrine peptides, microdialysis assay, 168, 166 rat cell aggregates cellular organization and topographical distribution, 168, 52 perifusion, 168, 57 preparation, 168, 50 cell culture primary, preparation, 168, 147 three-dimensional, preparation, 168, 48 cell type separation, 168, 62 patch clamp recording from cells identified by reverse hemolytic plaque assay, 168, 144 prolactin secretion, assay, 168,254 reverse hemolytic plaque assay, 168, 148 single cells hormone release, assay, 168, 266 hormone secretion, cell blot assay, 168, 327 intracellular calcium, assay, 168, 263 intracellular signaling, analysis, 168, 264

12 Antibiotics, s e e also s p e c i f i c a n t i b i o t i c s action, resistance, and working concentrations in E s c h e r i c h i a coli, 185, 22 Antibodies, s e e also Antisera, Immunoglobulins affinity purification, 194, 581 alprenolol, properties, 178, 266 amiloride, for characterization of transport systems, 191, 754 angiotensin I, cross-reacting material, reversed-phase HPLC, 168, 559 angiotensin II cross-reacting material, reversedphase HPLC, 168, 559 purification, 168, 546 antiallergen idiotopes on, analysis, 178, 150 immunoassay, 178, 155 preparation, 178, 151 -antigen interactions, protein footprinting analysis, 178, 746 - a n t i g e n - S t a p h y l o c o c c u s a u r e u s complexes antibody-antigen portion, solubilization, 182, 697 formation, 182, 695 for anti-Id antibody production, selection and purification, 178, 39 anti-idiotypic, s e e Anti-idiotypic antibodies anti-m6A in affinity chromatography of snRNPs U2 and U4/U6, 181, 244 purification, 181, 236 in purification of snRNP particles, 181,232 anti-m3G in affinity chromatography of nucleoplasmic snRNPs, 181, 241 in immunoprecipitation of snRNPs, 181, 83 purification, 181, 236 in purification of snRNP particles, 181, 232 anti-Sm, in immunoprecipitation of snRNPs, 181, 83 auto-, s e e Autoantibodies biotinylated, immobilization on avidincontaining column, 184, 301

13 biotinylation with biotinyl-N-hydroxysuccinamide, 184, 146, 316 via sugar moieties, 184, 162 catalytic design, 178, 551 immunoassay, 178, 542 production, 178, 531 purification, 178, 543 substrates and enzymatic assay, 178, 544 thiolated derivatization with spectroscopic probe, 178, 567 ester cleavage assays, 178, 565 fluorescence quenching binding assay, 178, 567 substrates, synthesis, 178. 564 thiolation, 178, 560 for chemiluminescent immunoassays. preparation, 184, 485 chimeric, production and properties, 178, 459 to complementary peptides, interaction with antigens, 168, 27 to DNA, immune suppression, 178, 422 engineered applications, 178, 498 expression in Escherichia coli, 178, 505 expression system requirements, 178, 501 production in yeast, 178, 486 structure, 178, 499 in evolutionary studies of nucleosomes, 170, 229 Fab fragments, engineered functional applications, 178, 498 expression in Escherichia coli, 178, 497 purification, 178, 512 structure, 178, 499 murine-human, secretion from Escherichia coli, 185, 184 production in Escherichia coli, 178, 477 yeast, 178, 486

Antibodies Fv fragments, functional expression in Escherichia coli, 178. 497 purification, 178, 512 to hepatitis B components, synthetic Tcell epitopes as immunogens for. 178, 634 to high-molecular-weight kininogen. preparation, 169, 280 to histidine-rich glycoprotein, in analysis of thrombin-stimulated releasate and plasma HRGP, 169, 273 histone, for study of nucleosomes. preparation, 170, 216, 251 and HPLC, in analysis of prohormone processing, 168, 517 idiotypic, for analysis of anti-ld antibodies to hormone receptors, 178. 196 immunoassay markers, sensitivity. 182, 705 for immunoelectron microscopy of nucleosomes application in indirect localization, 170, 190 preparation and selection, 170, 181 properties, 170, 184 for immunoprecipitation of small RNPs, preparation, 180, 471 for immunostaining, preparation, 194~ 617 insulin-like growth factor-ll receptor applications. 168, 320 characteristics, 168, 320 preparation and assay, 168, 318 JE, detection in sera, 184, 497 to leukotrienes, preparation, 187, 1 16 McPC603, variable domains, secretion. microinjection into amphibian oocytes, 170, 241 somatic cells, 170. 245 in monitoring protein folding and assembly, 191, 851 monoclonal, see Monoclonal antibodies monospecific, in analysis of cellular retinol- and retinoic acidbinding protein tissue distribution, 189, 315 vitamin A-regulated keratin expression, 190, 24

Antibodies to nonhistone proteins, preparation, 170, 218 -nucleosome complexes fixation, 170, 186 preparation, 170, 185 -nucleosome interactions, gel retardation study, 170, 230 to nucleosome nucleic acid components, preparation, 170, 219 to nucleosomes, preparation, 170, 219 -peptide interactions, size exclusion HPLC, 178, 122 to peptides preparation, 168, 7 site-specific, production, 178, 739 -peroxidase, and luminol, antigen detection, 184, 442 physalaemin, selection for studies of physalaemin-related molecules, 168, 448 to platelet PADGEM protein assay, 169, 314 preparation, 169, 312 -polyacrylamide beads, in solid-phase immunoassay of human IgA, 182, 709 poly(A) polymerase, effect on polyadenylation in vitro, 181, 67 polyclonal, s e e Polyclonal antibodies primary biotin-labeled, immunohistochemical application, 184, 361 for immunofluorescence studies in yeast, 194, 578 staining, 194, 618 probes of kgtl 1 recombinant DNA expression libraries, in isolation of yeast genes, 194, 233 prolactin, preparation and characterization, 178, 312 purified, biotinylation, 184, 146 for radioimmunoassay of 11-dehydrothromboxane B2, production, 187, 36 to receptors, production and characterization, 178, 179 with complementary peptides, 168, 25 recombinant, with antifibrin/plasminogen activator functions expression levels, 178, 526

14 production, 178, 516 purification and analysis, 178, 524 to ribonucleoproteins in mixed sera, relative concentrations, 180, 478 for RNA protection-immunoprecipitation experiments, 180, 457 secondary anti-IgG, conjugates with colloidal gold for immunostaining, 194, 619 for immunofluorescence studies in yeast, 194, 586 in selection assay for arachidonate 5lipoxygenase clone, 187, 494 site-specific, to synthetic rhinovirus receptor peptide antigens preparation, 178, 677 properties, 178, 683 purification, 178, 681 to snRNPs, effects on polyadenylation in vitro, 181, 67 streptavidin, preparation, 184, 435 synthetic peptide, in mapping sodium channel functional sites immunochemical antigen recognition, 178, 731 methods, 178, 720 overview, 178, 714 sodium current modification, assessment, 178, 736 trimethylguanosine, in RNA protection experiments, 181), 465 -tyrosine monooxygenase reactions, 168, 379 in vivo response, in assay of retinoid immunomodulatory activity, 190, 256 von Willebrand factor, titers, assay, 169, 161 Anticoagulants effects on blood and platelet preparations, 169, 147 in isolation of platelets from laboratory animals, 169, 29 Antidiuretic hormone, s e e Vasopressin Antigenic determinants, s e e Epitopes Antigenicity mycobacterial glycolipids, 179,236 peptides, potency enhancement, 178, 630 Antigens - a n t i b o d y - S t a p h y l o c o c c u s a u r e u s complexes

15 formation, 182, 695 solubilization of antigen-antibody portion, 182, 697 -avidin conjugates, construction, 184, 645 binding of idiotope, inhibition by anti-Id antibodies, 178, 86 -biotin conjugates, construction, 184, 647 biotin-labeled, precipitation, 184, 518 biotinylation, 184, 473 blotted, labeling, 184, 424 carcinoembryonic assay in serum, 184, 507 biotinylated, in epitope analysis, 184, 524 conformational changes, detection, 178, 761 HBcAg specific T cells, priming of antibody production to HBsAg, 178, 652 synthetic peptides, priming of T-helper activity in v i v o , 178, 652 HBsAg antibodies, induction with synthetic Tcell epitopes, 178, 634 pre-S(1) region, T-cell epitopes fine specificity, 178,646 identification, 178, 646 priming of T-helper activity for antibody to B epitopes, 178, 649 pre-S(2) region, T-cell recognition, fine specificity, 178, 644 human B- and T-cell, immunocytochemical detection, 184, 363 immunoassay markers, sensitivity, 182, 7O5 on immunodots, luminescent detection, 184, 442 interactions with antibodies to complementary peptides, 168, 27 protein footprinting analysis, 178, 746 size-exclusion HPLC binary complexes, 178, 108 ternary complexes, 178, 116 neo-, s e e Neoantigens nonabundant, detection in immunofluorescence studies, 194, 593

Anti-idiotypic antibodies -peptide system, high-density multiple, for antipeptide antibody preparation, 168, 7 for polyclonal antibody preparation, purification and preparation. 182. 663 radiolabeled, immunoadsorption. 182. 692 related protein segments, identification by hydrophilicity plotting, 178. 571 synthetic peptide in distinguishing HIV-1 and HIV-2 infections, 178, 709 ELISA with, 178, 705 in immunoassays, advantages and limitations, 178, 695 rhinovirus receptor-related, sitespecific antibodies, production. 178, 676 tumor, anti-idiotype antibodies to. 178, 15 on Western blots detection with streptavidin-enzyme complexes, 184, 437 luminescent detection, 184, 442 Anti-idiotypic antibodies acetylcholine receptor immunotoxin with, preparation and testing, 178, 453 preparation, 178, 452 adenosine deaminase-binding protein in Id-anti-Id immunoassay, 178, 174 production, 178, 173 /3-adrenergic catecholamine receptor, responses in mouse, 178, 272 rabbit, 178, 269 /3-adrenergic receptor, 178, 25 angiotensin II receptor, 178, 25 antiallergen antibody, preparation, 178, 154 anti-DNA antibody, conjugate with neocarzinostatin immune suppression of anti-DNA antibody production, 178, 422 preparation, 178, 424 properties, 178, 427 antireceptor antibody, preparation and characterization, 178, 184 associated networks. 178, 3

Anti-idiotypic antibodies hemitoxin- or holotoxin-modified, in treatment of B-cell neoplasms, 178, 356 -idiotope reaction, blocking by antibodies, 178, 87 and idiotype complexes, immunoelectron microscopy, 178, 130 in immunocytochemical studies of receptors, 178, 213 insulin receptor, 178, 23 -membrane receptor interactions, 178, 179 modulation of anti-AChR response, 178, 444 molecular modeling, 178, 103 morphine, production, properties, and antiopiate receptor activity, 178,222 to murine B-cell lymphoma -drug conjugates pharmacological activity, 178, 374 preparation, 178, 371 production, 178, 370 nicotinic acetylcholine receptor, 178, 24 opioid receptor, production and characterization, 178, 243 with predefined specificity, generation with cRNA, 178, 63 probes of hormone receptor structure and function, 178, 191 prolactin receptor binding to membranes, assay, 178, 315 characteristics, 178, 316 generation, 178, 313 inhibition of prolactin binding to membranes, 178, 316 purified, assay, 178, 162 reagent production and characterization, 178, 36 reovirus type 3 HA cell receptor in characterization of viral receptor, 178, 325 polyclonal antisera with, development and characterization, 178, 322 sequence determination with Ig mRNA, 178, 94 serological detection, 178, 74 for small ligands, production and characterization, 178, 49 specificity, characterization, 178, 81 in structure-function studies of protein active sites, 178, 163

16 substance P immunological characterization, 178, 283 production, 178, 276 properties, 178, 292 substance P receptor, 178, 24 thyroid-stimulating hormone receptor, 178, 26 in treatment of experimental autoimmune myasthenia gravis, 178,433 lymphoid tumors, 178, 341 vaccines in infectious diseases, 178, 7 Antioxidants assay principles, 186, 303 effects on chemiluminescence, 186, 608 arAntiplasmin platelet, radioimmunoassay, 169, 298 Antisera, s e e a l s o Antibodies for analysis of idiotopes on antiallergen antibodies, 178, 152 angiotensin II affinity chromatography, 168, 547 purified, characterization, 168, 549 anti-idiotypic polyclonal, to reovirus type 3 hemagglutinin in characterization of viral receptor on cells, 178, 325 development and characterization, 178, 322 for arachidonic acid metabolites immobilization, 187, 65 preparation, 187, 64 to cellular retinoid- and retinoic acidbinding proteins, preparation, 189, 276 hnRNP protein, preparation, 181, 320 in identification of leukotriene A4 hydrolase cDNA clones, 187, 486 for immunoassays, 182, 702 for immunoneutralization,interfering substances, 168, 512 for inhibin detection and purification, 168, 588 for isolation of yeast m n n mutants, 185, 446 polyclonal, interactions with peptides, size exclusion HPLC analysis, 1"/8, 124 for preparation of immunological probes for nucleosomes, 170, 215

17 procholecystokinin, development, 168, 3 protein reactivity, relationship to/3-turn incidence, 178, 592 for reverse hemolytic plaque assay, 168, 146 serum retinol-binding protein, preparation, 189, 271 tyrosine hydroxylase, preparation, 168, 376 whole, biotinylation, 184, 148 c~rAntitrypsin detection on Western blots, 184, 441 Antitumor drugs -anti-Id antibody conjugates for treatment of murine B-cell tumors pharmacological activity, 178, 374 preparation, 178, 370 efficacy, testing with ascites tumors, 171, 601 Aorta porcine, smooth muscle membranes, preparation, 187, 400 rabbit, subendothelium, preparation, 169.48 Aphids stylets, in collection of sieve tube sap, 174, 305 Apolar cells chloride channel blocker, description, 191, 801 Apoplast hydraulic properties, 174, 198 water storage capacity, 174, 209 Arachidonate activation of Ca 2~ channels, 191, 683 intracellular Ca -'+ mobilization, 191, 684 analysis in isolated perfused rat lung, 187, 599 compartmentalization, 191, 685 disposition on stimulation of phosphatidylinositol breakdown, 191, 688 formation from diacylglycerol, inhibition by RHC 80267, 191, 690 3H-labeled, labeling of endothelial cells, 187, 532 liberation from phospholipids, mechanisms. 191, 677 metabolism by ocular cytochrome P450, 187, 372

Arachidonic acid in parietal cells, 187, 505 role in protein kinase C activation. 191, 684 metabolites, mitogenic action, 191,682 phosphoglycerides with, separation and characterization, 187, 157 radiolabeled, transformation assay. 187. 338 role in phospholipase C activity. 191. 683 Arachidonate-CoA ligase assay, 187, 238 defective mutant cell line, production. 187, 241 separation from long-chain-fatty-acidCoA ligase, 187, 240 solubilization, 187, 238 Arachidonate 5-1ipoxygenase activation in bone marrow-derived mast cells, differentiation from exocytosis, 187, 518 expression in insect cells with baculovirus vector, 187, 496 human leukocyte isolation and characterization. 187, 312 stimulatory factors, preparation, 187, 316 molecular biology and cloning, 187, 491 monoclonal antibodies, preparation. 187. 341 peptide sequence analysis, 187, 492 polyclonal antibody preparation. 187. 493 porcine leukocyte immunoaffinity purification, 187, 338 properties, 187, 342 potato assay, 187, 297 properties, 187, 301 purification, 187, 300 potato tuber assay, 187, 269 properties, 187, 273 purification, 187, 268 recombinant characterization, 187, 500 transfer vector, construction, 187, 497 Arachidonic acid effect on citrated platelet-rich plasma. 169, 132

Arachidonic acid epoxidation, 187, 360 3H-labeled, hydrolysis, in assay of phospholipase A2, 187, 216 lipoxygenase products, HPLC, 187, 98 metabolism in cultured cells, 187, 535 endothelial cells, 187, 531 human platelets, 187, 589 isolated and cultured mTALH cells, 187, 369 lymphocyte preparations, 187, 582 platelet preparations, 187, 582 metabolites HPLC, 187, 378 immunoaffinity purification-chromatographic analysis, 187, 62 reversed-phase HPLC, 187, 370 stimulated release from mTALH cells, 187, 371 TLC, 187, 377 oxidation by cytochrome P450 enzymes, 187, 390 oxidative metabolism in bone marrowderived mast cells, 187, 514 oxygenation by prostaglandin H synthase, 186, 285 Arginine in anion-transport protein functionally essential residues, location, 173, 483 reaction with phenylglyoxal, 173, 467 exofacial modification, 173, 479, 483 intracellular modification, 173, 487 spin system identification, 177, 139 transport in intact isolated vacuoles from yeast and Neurospora, 174, 515 Arginine kinase bound reactants and products, equilibrium constant determination, 177, 362 interconversion rates, NMR analysis, 176, 297 3~p NMR, simulated and experimental spectra, comparison, 176, 299 Arginine permease yeast vacuolar, characterization, 174, 504 Arginine vasopressin receptors immunocytochemical images, 178,219 immunocytochemical studies with antiidiotypic antibodies, 178, 216

18 Aromatase granulosa cell, in bioassay for folliclestimulating hormone, 168,414 Aroxyl radicals flavonoid, formation and decay, rate constants, 186, 349 intermediates, transient spectra, 186, 351 kinetics and mechanisms, 186, 97 reaction in solution, rate constants, 186, 114 Arteries, see also specific arteries human, subendothelium preparation, 169, 47 segments, postfixation, dehydration, and Epon embedding, 169, 62 Arterioles renal blood collection, 191, 121 resistances, 191, 125 Arthritis rheumatoid, see Rheumatoid arthritis Arthrobacter A. globiformis, purification of

glucose-6-phosphate dehydrogenase, 188, 341 6-phosphogluconate dehydrogenase, 188, 343 Pl, purification of formaldehyde dehydrogenase, 188, 319 methylamine oxidase, 188, 229 transaldolase isoenzymes, 188, 407 Arylazides generated nitrenes, properties, 172, 637 Asci yeast micromanipulationand dissection, 194, 21 wall digestion with snail juice, 194, 32 Ascites monoclonal antibody-containing,electrophoresis, 178, 56 Ascites fluid murine, tumor cells from, membrane potential assay, 172, 95 purification of monoclonal antibody to biotin, 184, 117 Ascorbate autoxidation, 186, 125 -Cu 2+ system, radical damage, assay, 186, 167

19 in detection of catalytic metals in simple buffers, 186, 125 Ascorbic acid radioprotective effects in mice, 186, 686 Ascospores yeast isolation of RNA, 194, 401 pure and viable, preparation, 194, 125 separation, 194, 34 Asialofetuin -colloidal gold complexes, preparation, 179, 114 Asparagine linked carbohydrates, attachment site and structural class in glycoproteins, 193, 501 linked oligosaccharides, structural studies, 193, 672 in proteins, analysis, 182, 600 spin system identification, 177, 138 Asparagine synthetase, see Aspartateammonia ligase Aspartate /3-carboxylation, 182, 635 carboxyl methylation, 182, 632 Aspartate-ammonia ligase in heterologous gene selection and coamplification in mammalian cells, 185, 557 Aspartate transcarbamoylase -carbamoyl-phosphate synthase-dihydroorotase, in gene selection and coamplification, 185, 550 Aspartic acid spin system identification, 177, 138 Aspartic proteinases phylogenetic relationships, 183, 682 Aspergillus phoenicis

purification of exo-al--.2-mannosidase, 185, 463 Aspirin treated endothelial cell monolayers, platelet adherence, 169, 75 treatment of platelets and endothelial cells, 187, 588 Atherosclerosis role of oxidants, 186, 62 Atomic absorption spectroscopy in assay of total Ca 2+ in hepatocytes, 173, 540

ATPase Atomic nuclei NMR-insensitive, detection. 176. 134 selectively excited, spin-lattice relaxation, 176, 188 spin 3/2, undergoing exchange, multiple relaxations, 171, 292 Atomic weight selected elements, 193, 870 ATP (adenosine 5'-triphosphate) -ADP exchange in mitochondria, characterization, 171, 12 caged, synthesis and properties, 172. 289 cofactor role in enzymatic addition of guanylate to tRNA H~-',181. 459 driven CI- pump in Acetabularia electrical properties. 174, 493 electrogenic, demonstration. 174, 492 kinetic models. 174, 498 driven H ÷ transport in renal cortical vesicles, 191, 505 effects on vacuolar arginine permease. 174. 512 vesicular glutamate uptake, 174, 19 hydrolysis, dependence of light-induced hyperpolarization, 174, 487 levels in erythrocytes control, 173. 105 modification, 173, 86 permeabilization of mammalian cells to macromolecules, 171, 860 32p-labeled, labeling of crosslinking sites in DNA-protein complexes. 170. 406 31p NMR line shapes, simulated and experimental, comparison, 176. 291 synthesis by chloride pump reversal in vivo. 174, 501 ATPase assay, 192, 155 Ca 2+_ coupling rules, 171, 156 in one-step inside-out vesicles, assay. 173, 373 purification by transport-specific fractionation, 172, 34 rabbit muscle sarcoplasmic reticulum, crosslinking, 172, 606 reconstitution, 172, 35 distribution in gastric oxyntic cells, 192, 158

ATPase n

+.

CO2-induced exocytotic insertion, regulation of urinary epithelial H + transport, 172, 54 coupling rules, 171, 158 in endocytotic vesicles, assay, 191, 509, 517 proton translocation in Neurospora crassa, assay, 174, 668,674 yeast mitochondrial, assay, 194, 658 in plasma membrane, assay, 194, 658 vacuolar, assay, 194, 656 H+,K ÷blockers, effects on H ÷ transport and ATPase, 191, 734 interactions with reversible K+-site antagonists, 191, 737 membranes with gastric sources, 191, 734 isolation from gastric oxyntic cell, 192, 151 pH gradient formation, assay, 192, 161 myosin site biotinylation, 184, 412 visualization, 184, 409 Na+,K +active site phosphorylation by inorganic phosphate, assay, 173, 688 binding of cardiac glycosides, 173, 678 in bioassay of arachidonic acid metabolites, 187, 378 biochemical assays, 191, 543 characterization with amiloride and analogs, 191, 744 in cultured heart cells characterization, 173, 639 effects of ouabain and low K+ levels, 173, 651 inhibition, correlation with glycoside receptor occupation, 173, 649 number of molecules, determination, 173, 643 relationship to positive inotropic action of glycosides, 173, 655 electrolyte transport studies, 192, 381 in erythrocyte inside-out vesicles assay, 173, 372

20 stoichiometry and coupling, 173, 377 in isolated cells, assay, 173, 676 in muscle, assay, 173, 695 in muscle fibers, stoichiometric ratio, 173, 707 in parietal cells, assay, 192, 78 rate measurement with radioactive tracers, 173, 700 transport characteristics in erythrocytes, 173, 80 turnover rate, measurement, 173, 712 in turtle colon basolateral membrane, analysis, 192, 740 vacuolar-type, in goblet cell apical membrane assay, 192, 613 purification, 192, 612 ATP synthase F~F0-, proton-translocating portion F0, photochemical labeling, 172, 663 Atrial natriuretic factor binding to isolated nephron segments, microassay, 191, 315 Atrial natriuretic factor receptors characterization with amiloride, 191, 747 Autoantibodies depletion of small RNPs from extracts, 180, 480 in immunoprecipitation of RNPs, 180, 468 reactions with nonmammalian small RNPs, 180, 479 small RNPs recognized by, 180, 470 Autofluorescence correction for assay of cytosolic free Ca 2÷, 172, 247 Autographica californica nuclear polyhedrosis virus -Spodoptera frugiperda Sf9 cell system construction, 182, 119 expression of mammalian proteins, 182, 117 recombinant viral genes and gene products, detection reagents, 182, 124 Autography fibrin, in assay of oocyte tissue plasminogen activator, 168, 424

21 Autoimmune diseases ramifications of molecular mimicry concept, 178, 19 Autoimmune responses to acetylcholine receptor, inhibition with idiotype-specific immunotoxins, 178, 448 in myasthenia gravis, inhibition, 178, 450 Autoimmunity ramifications of molecular mimicry concept, 178, 19 Autolysis yeast cells, 182, 159 Autonomic nervous system animal models, 168, 434 functional assessment, 168, 432, 442 peptide regulation, 168,431 Autoradiography in analysis of UV-induced RNA-RNA crosslinks, 180, 422 in detection of protein degradation in intact cells, 186, 498 film, in quantitation of in situ hybridization, 168, 749 in situ hybridization histochemistry, 168, 809 and immunohistochemistry, in analysis of vitamin A-regulated keratin expression, 190, 25 micro-, see Microautoradiography Northern and dot blots, 168, 407 O-linked oligosaccharides from yeast glycoproteins, 194, 696 in one-dimensional electrophoresis of nucleosomes, 170, 121 quantitative, and in situ hybridization, data analysis, 168, 825, 840 retinoid-binding proteins, 189, 303 signal quantitation by image analysis, 168,410 silver stain quenching, 182, 486 in in situ hybridization histochemistry detection of neuropeptide mRNA, 168, 705 at high-resolution, 168, 768, 776 with markers of neuronal connectivity, 168, 787 tissue sections after in situ hybridization of retinoid-binding protein mRNA, 189, 294

Avidin in vitro, in binding site analysis of dif-

fuse neuroendocrine system. 168, 8O6 Autoxidation ascorbate, 186, 125 hematoxylin in assay of superoxide dismutase. 186, 22O transition metal effects, 186, 223 iron, continuous assay, 186, 462 Avidin affinity purification, 184, 53 -agarose, synthesis, 184, 297 -2-anilinonaphthalene-6-sulfonic acid titration by biotin, 184, 235 titration of biotin, 184, 239 -antigen conjugates, construction, 184. 645 attachment to biotinylated site of heavy meromyosin, 184, 413 -biotin interaction, ftuorometric assay, 184, 234 -biotin system blot staining, 184, 425 composite applications, 184, 618 dissociation constants, 184, 60 as electron microscopic probe, 184, 415 ELISA enhancement, 184, 537 in enzyme immunoassays, 18,1, 469 as gene probes, 184, 560 -horseradish peroxidase, in immunoassay of HIV core proteins, 184, 556 in immunohistochemistry, 184, 357 in isolation of biologically active compounds, 184, 243 gonadotropin-releasing hormone receptors, 184, 287 labeling of blotted antigens and receptors, 184, 424 in localization of ganglioside GMt, 184, 405 gonadotropin-releasing hormone receptors, 184, 395 lectin receptors, 184, 388 mediated immunoassay, overview. 184, 467

Avidin

22

related technology preparation, 184, 225 applications, literature survey, 184, 14 immobilization, 184, 185 overview, 184, 5 in immunocytochemical detection of in screening epitope specificities of human T- and B-cell antigens, 184, monoclonal antibodies, 184, 507 363 -biotinylated enzyme complexes, prepain isolation of ration, 184, 181 ACTH-binding proteins, 184, 265 -biotinylglycan neoglycoproteins insulin receptors, 184, 249 applications, 184, 658 labeling systems, advantages, 184, 404 preparation and properties, 184, 656 linked complement C3b oligomers with biotinyl ligand displacement kinetics, enhanced affinity for receptors, 184, 184, 64 626 chemical modification, 184, 476 monovalent cloning and expression in Escherichia affinity columns, chromatography coli, 184, 70 with, 184, 199 colorimetric enzyme assay, 184,217 -agarose column with, immobilization of biotinycharacteristics, 184, 196 lated antibodies, 184, 301 preparation, 184, 195 coupling to nonglycosylated enzymes, 184, 475 affinity chromatography, 184, 69 erythrocytes, 184, 317 crystal forms, 184, 92 derivatives, affinity chromatography, properties, 184, 70 184, 171 nonspecific binding egg-white comparison with streptavidin, 184, crystal forms, 184, 90 206 succinylation, 184, 383 prevention, 184, 201 as electron microscopic probe, 184, 413 -penicillinase conjugates, preparation, enzymatic and radioactive assays, 184, 184, 179 208 -peroxidase conjugates, and luminol, -ferritin conjugates antigen detection, 184, 442 preparation, 184, 177 -phycobiliprotein conjugate, preparain ultrastructural localization of retinal tion, 184, 188 photoreceptor proteins, 184, 370 -phycoerythrin conjugate, preparation, fluorescein-derivatized, preparation, 184, 184, 193 174 polymers, characteristics, 184, 60 general properties, 184, 56 selective precipitation of biotin-labeled -gold conjugates antigens or monoclonal antibodies, with biotinyl ligand, in visualization of 184, 518 intracellular protein trafficking, -Sepharose, in identification of comple184, 384 ment C3 acceptors, 184, 624 colloid preparation, 184, 381 -toxin conjugates, preparation, 184, 178 historical overview, 184, 3 tritiation, 184, 176 -horseradish peroxidase, in detection of Avocado sunblotch viroid biotinylated proteins, 184, 435 cDNA clone preparation, 181, 584 leukocyte surface proteins, 184, 431 RNA self-cleavage, characterization, hydrazide derivatives, preparation, 184, 181, 583 184 Azides 125I-labeled 3H-labeled, preparation, 191, 765 based sequential solid-phase assay of as membrane perturbants and photoaffinbiotin, 184, 224 ity labeling agents, 191,767

23 p-Azidobenzylphloretin apparent affinity for sugar transporter, 191, 771 3H-labeled, photolabeling of red cells, 191, 778 interaction with anion transporter, 191, 776 synthesis, 191, 759 p-Azidobenzylphlorizin apparent affinity for sugar transporter, 191, 771 effect on salt and water flux, 191, 776 interaction with anion transporter, 191. 776 synthesis, 191, 761 3'-(4-Azidobenzyl)-2',4',6',4-tetrahydroxydihydrochalcone, see p-Azidobenzylphloretin 5'-[(4-Azidophenacyl)thio]phosphoryladenosine synthesis, 180, 395 5'-[(4-Azidophenacyl)thio]phosphorylguanosine synthesis, 180, 395 4-Azidophlorizin synthesis, 191, 764 4-Azido-p-phlorizin synthesis, 191, 765 Aziridine heteroatom analog of epoxyeicosatrienoic acids, preparation, 187, 364 2,2'-Azobis(2-amidinopropane) dihydrochloride generation of water- or lipid-soluble peroxyl radicals, 186, 100 2,2'-Azobis(2,4-dimethylvaleronitrile) generation of water- or lipid-soluble peroxyl radicals, 186, 100 Azo compounds oxidation of tocopherols, 186, 199 (7,7-Azo-3a, 12c~-dihydroxy-5/3-cholan-24oyl)-2-amino[l,2,-3H]ethanesulfonic acid synthesis, 174, 26 Azurin assay, 188, 285 properties, 188, 288 purification from organism 4025, 188, 285

Bacteria B Bacillus B. circulans, purification of endo-al-~6-

mannanase, 185, 461 B. macerans, purification of protocate-

chuate 2,3-dioxygenase, 188.97 B. subtilis

culture media, optimization for heterologous protein secretion. 185. 222 expression of heterologous genes dual-plasmid repressible system, 185, 208 inducible system employing integrated lacl gene, 185, 211 lac-based inducible system, 185. 203 overview, 185, 199 plasmid pREP9-based system. 185. 205 expression systems, overview, 185. 6 plasmid instability in, 185, 200 protein secretion, overview. 185. 199 purification of ribosomal proteins BII6 and ELI6. 181, 373 RNase M5, 181, 372 regulated expression of active HIV-I reverse transcriptase, 185, 210 regulatory genes, inducible expression, 185, 223 secreted proteases, effects on protein stability, 185, 200 secretion of heterologous proteins. 185, 214 spoOA gene, placement under inducible control. 185, 225 sporulation, control by IPTG, 185, 225 synthesis of precursors for heterologous proteins, demonstration. 185, 220 transformation with integrative plasraids, 185, 225 C1, purification of 3-hexulose-6-phosphate synthase, 188. 393 methanol dehydrogenase, 188, 224 Bacteria, see also specific bacteria amino acid transport, 177, 59 associated network epitopes, 178, 12

Bacteria capsular polysaccharides, polyacrylamide gel electrophoresis, 179, 104 cloning of yeast mitochondrial DNA, 194, 159 genomic divergence through gene rearrangement, 183, 428 gram-negative, enzymatic lysis, 182, 148 gram-positive, enzymatic lysis, 182, 147 growth, suppression in NMR samples, 176, 78 isolation of pyrroloquinoline quinone, 188, 261 methylotrophic growth, 188, 351 purification of citrate synthase, 188, 351 serine pathway, preparation of crude extracts, 188, 361 priSe plasmid-bearing, T4 lysozyme production, 177, 72 purification of exocellular superoxide dismutase, 186, 249 strains auxotrophic for amino acids, 177, 55 superoxide dismutase assay in colonies, 186, 240 whole cells, 186, 237 Bacteriophages CE6 delivery of T7 RNA polymerase to cell, 185, 65 infection of Escherichia coli for expression of target DNA, 185, 83 stocks, preparation, 185, 83 DE3, in "1"7RNA polymerase-directed gene expression in Escherichia coli, 185, 63 hgt 11 in isolation of yeast genes, 194, 230 recombinants, expression of foreign DNA, 194, 230 MI3 growth in dut-ung- host, 185, 605 uracil-containingtemplates, in saturation mutagenesis, 185, 599 T4, pin gene, inhibition of proteolysis in Escherichia coli, 185, 127 T4td-, in analysis of RNA splicing in Escherichia coli, 181, 523

24 Bacteriorhodopsin in coreconstitution of CI- channels, 191, 580 interaction with diheptanoylphosphatidylcholine, 171, 272 photochemical labeling, 172, 666 reconstitution into lipid vesicles, 171, 267 Bacterium W3A1 purification of electron-transfer flavoproteins, 188, 309 methylamine dehydrogenase, 188, 243 trimethylamine dehydrogenase, 188, 253 Baculovirus in expression of arachidonate 5-1ipoxygenase in insect cells, 187, 496 transfer vector for recombinant 5-1ipoxygenase, construction, 187, 497 Barbital buffer stock solutions, 182, 35 Barium in determination of ion transport mechanisms, 171, 621 Bases, see also specific bases BrOnsted, reagents for chemical ionization, 193, 19 ions in mass spectra of trimethylsilyl derivatives of nucleosides, 193, 831 loading mechanisms, identification in parietal cells, 192, 54 long-chain, from membrane lipids HPLC, 172, 558 LC-MS, 172, 567 nucleic acid, trimethylsilyl derivatives, preparation, 193, 792 weak accumulation by parietal cells, determination, 192, 104 and weak acids, equilibrium distribution, 172, 79 Basicity free radicals, 186, 92 BATGEN program prediction of optimal and suboptimal RNA secondary structure, 183, 286, 304

25 B cells epitopes based peptides, addition of Th-cell epitopes, 178, 661 computer prediction, 178, 586 in HBsAg anti-HBs production to, T-help priming, 178, 649 pre-S(l) sequence identification, 178, 641 pre-S(2) region identification, 178, 639 human, antigens, immunocytochemical detection, 184, 363 isolated cell membranes, phosphoinositide hydrolysis, assay, 168, 338 murine, experimental tumors, treatment with anti-Id antibody-drug conjugates, 178, 369 neoplasms involving, treatment with idiotype-specific immunotoxins, 178, 356 tumors involving, treatment with antiidiotypic antibodies, 178, 341 Beads glass, see Glass beads impregnation with all-trans-retinoic acid, 190, 204 polyacrylamide, see Polyacrylamide beads retinoic acid-impregnated, implantation in embryonic tissues, 190, 205 Beauvericin ion carrier in planar bilayers, relaxation and noise analyses, 171, 274 Beetle hemolymph, lectin purification, 179, 331 Benz2 in control of red cell Ca:+ content, 173, 105 cis-Benzene dihydrodiol dehydrogenase assay, 188, 134 properties, 188, 137 purification from Pseudornonas putida NCIB 12190, 188, 136 Benzene dioxygenase detection by antibody probing, 188, 55 ferredoxin component, purification, 188, 59

Bicarbonate oxygen electrode assay, 188, 53 purification, 188, 56 reductase component, purification, 188. 58 spectrophotometric assay, 188.54 terminal dioxygenase properties, 188, 59 purification, 188, 58 subunit preparation, 188, 59 Benzoate-CoA ligase assay, 188, 154 properties, 188, 157 purification from Rhodopseudomonas palustris, 188, 156 Benzoic anhydride per-O-benzoylation of glycosphingolipids, 172, 547 p-Benzoquinones half-wave reduction potentials, substituent effects, 186, 187 HPLC with electrochemical detection, 186, 180 Benzoylation diacylglycerides, 187, 207 Benzoyl chloride per-O,N-benzoylation of glycosphingolipids, 172, 544 Benzoyl-diacylglyceride molecular species separation, 187, 210 subclass separation, 187, 209 Benzylamine oxidase assay, 188, 427 properties, 188,433 purification from Candida boidinii, 188, 429 Benzylamine/putrescine oxidase assay, 188, 427 properties, 188, 433 purification from Pichia pastoris, 188, 429 Bicarbonate -carbonate buffer stock solutions, 182, 37 -CI- exchange detection and analysis, 173, 344 driven C1- uptake by intestinal membrane vesicles, 192, 407 -CO2 solutions, pH control, 192, 42 -Fe(IV) complex, generation in aqueous solution, 186, 111

Bicarbonate secretion from duodenal mucosa, 192, 147 gastric mucosa, 192, 141 transport across nutrient membrane, mechanistic analysis, 171, 622 in pancreatic duct, analysis, 192, 269 Bicinchoninic acid in colorimetric assay of proteins, 182, 60 Bile canalicular motility, 192, 509 canalicular secretion, 192, 508 glutathione in, assay limitations, 173, 531 glutathione disulfide excretion into, as indicator of oxidative stress, 173, 531 Bile acids carrier-mediated transport, characteristics, 192, 531 hepatic transport mechanisms, 192, 517 physical properties, 192, 518 Bile pigments antioxidant activities, 186, 301 isocratic HPLC, 186, 309 Bilirubin antioxidant activities, 186, 301 isocratic HPLC, 186, 309 Bilirubin transporter isolation from hepatic plasma membrane, 174, 50 Bilitranslocase binding assay, 174, 51 isolation from hepatic plasma membrane, 174, 50 Biliverdin antioxidant activities, 186, 301 isocratic HPLC, 186, 309 Binary signals pseudo-random, in measurement of transepithelial impedance, 171, 631 Binding assay anti-idiotypic antibodies to anti-AChR antibodies, 178, 438 antiprolactin, to membrane, 178, 315 antisubstance P, to physiological receptors, 178, 292 bilitranslocase, 174, 51 cellular retinoic acid-binding proteins I and II, 189, 360

26 competition, morphine:Abl-Ab2, 178, 238 DNA, 182,527 fluorescence quenching, with fluorescein-thiolated Fab adduct, 178, 567 glucosidase I, 179, 456 glucosidase II, 179, 461 gonadotropin-releasing hormone to membranal and solubilized receptors, 184, 289 hormones and drugs to isolated nephron segments, 191, 315 inhibition studies, s e e Binding inhibition assay insulin, 184, 253 leukotriene D4 receptors, 187, 417 macromolecules to neutrophils, 186, 581 mannosidase I, 179, 464 mannosidase II, 179, 468 monoclonal antibody-mutant diphtheria toxin conjugates, 178,419 in monolayer cell cultures, devices for, design, 171, 133 neurohypophyseal peptides, tracer effects, 168, 581 opioid receptors, 178, 246 polyadenylate-binding proteins, 181, 333 prolactin binding to membrane fractions, 178, 315 with radiolabeled amiloride analogs, 191, 751 radioligand, s e e Radioligand binding assay 1 1 - c i s - r e t i n a l d e h y d e to cellular retinaldehyde-binding protein, 189, 370 retinol-binding proteins, 190, 142 solid-phase, with complementary peptides, 168, 23 steroid-receptor complexes to isolated nuclei, 191, 532 sulfidopeptide leukotriene receptors, 187, 421 in testing production of anti-idiotype antibodies to hormone receptors, 178, 201 Binding constants biotinylated ligand-immobilized streptavidin interaction, 184, 633 internal and external, paramagnetic probes, 172, 338

27 ions at membranous polypeptide sites, NMR determination, 171, 286 Binding energy role in active transport and other coupled vectorial processes, 171, 145 Binding inhibition assay anti-idiotypic antibody inhibition of anti-AChR antibody-receptor binding, 178, 439 hormone receptor binding by polyclonal idiotypes, 178, 202 idiotope binding to antigen, 178, 86 prolactin binding to membranes, 178, 316 Bioassay arachidonic acid metabolites, 187, 378 diuretic hormones, 191, 178 eicosanoid action in isolated perfused rat lung, 187, 605 follicle-stimulating hormone, 168, 414 gastrin-releasing peptides, 168, 663 inhibin in vitro, 168, 602 paf-acether, 187, 125 platelet-activating factor, 187, 130 position-dependent differences in blastema cell affinity along regenerating limb, 190, 198 pyrroloquinoline quinone, 188, 270 Biocytinamide and carboxypeptidase Y, C-terminal biotinylation of proteins, 184, 160 Biocytin hydrazide biotinylation of glycoproteins, 184, 156 preparation, 184, 130 Biocytin hydrochloride preparation, 184, 128 Biodegradation polycyclic aromatic hydrocarbons by Mycobacterium, 188, 148 Biopsy skin, free radical reduction at epidermal surface, 186, 674 Bio-19-SS-dUTP incorporation into replicating SV40 chromosomes, 170, 45 synthesis, 170, 42; 184, 585 Biotin -antigen conjugates, construction, 184, 647

Biotin -avidin interaction, fluorometric assay, 184, 234 -avidin system blot staining, 184, 425 composite applications, 184, 618 dissociation constants, 184, 60 ELISA enhancement, 184, 537 in enzyme immunoassays, 184, 469 as gene probes, 184. 560 -horseradish peroxidase, in immunoassay of HIV core proteins. 184, 556 in immunohistochemistry, 184. 357 in isolation of biologically active compounds, 184, 243 gonadotropin-releasing hormone receptors, 184, 287 labeling of blotted antigens and receptors, 184, 424 in localization of ganglioside Gut, 184, 405 gonadotropin-releasing hormone receptors, 184, 395 lectin receptors, 184, 388 mediated immunoassay, overview. 184, 467 related technology applications, literature survey, 184, 14 overview, 184, 5 in screening epitope specificities of monoclonal antibodies, 184, 507 binding proteins in egg-yolk, 184, 99 homology around Trp residues, 184, 101 nonavidin types, 184, 93 in nucleus, 184. 100 overview, 184, 49 binding to streptavidin, blot assay, 184, 85 -cellulose conjugates, synthesis, 170.49 colorimetric enzyme assay, 184, 217 coupling to ~zSI-labeled histamine, 184, 213 125I-labeled histidine, 184, 217 direct incorporation into DNA, 184, 608 endogenous, nonspecific binding of avidin, prevention. 184, 201

Biotin enzymatic assays, 184, 208 historical overview, 184, 3 hydrazide derivatives, in antibody biotinylation via sugar moieties, 184, 163 in immunocytochemical detection of human T- and B-cell antigens, 184, 363 iodinated imidazole derivatives, synthesis, 184, 211 labeled antigens and monoclonal antibodies, selective precipitation, 184, 518 labeled chemically cleavable nucleotide analogs, synthesis, 184, 584 labeled primary antibodies, immunohistochemical application, 184, 361 monoclonal antibody production and characterization, 184, III purification from ascites fluid, 184, ll7 -nucleotide analogs applications, 184, 576 incorporation into DNA, 184, 572 synthesis and characterization, 184, 562 -phycobiliprotein conjugates, preparation, 184, 188 -phycoerythrin conjugates, preparation, 184, 193 radioactive assays, 184, 208 reagents with, preparation, 184, 123 sequential solid-phase assay based on ~25I-labeledavidin, 184, 224 -streptavidin conjugate in colorimetric detection of DNAsequencing bands, 184, 614 in ELISA of herpes simplex virus, 184, 547 in isolation of splicing complexes, 184, 324 titration, 184, 239 titration of 2-anilinonaphthalene-6sulfonic acid-avidin complex, 184, 235 5-[(N-Biotin-amido)hexanoamido-ethyl- 1,3dithiopropionyl-3-aminoallyl]-2'-deoxyuridine 5'-triphosphate, see Bio-19-SSdUTP

28 Biotin aminocaproylhydrazide biotinylation of antibodies via sugar moieties, 184, 163 Biotin holocarboxylase synthetase overview, 184, 94 Biotin hydrazide interaction with DNA, 184, 608 preparation, 184, 127 Biotinidase assays, 184, 97, 104 biotin-bindingassay, 184, 97 catalytic properties, 184, 108 inhibition, 184, I l0 overview, 184, 95 physical properties, 184, 108 specificity, 184, 109 stability, 184, I 11 Biotinylalprenolol derivatives, preparation and characterization, 184, 660 Biotinyl-N-e-aminocaproic acid preparation, 184, 128 Biotinyl-N-e-aminocaproyl-N-hydroxysuccinimide ester preparation, 184, 129 Biotinyl-N-e-aminocaproyl-p-nitrophenyl ester preparation, 184, 130 Biotinylation ACTH-binding proteins, 184, 264 alkaline phosphatase, 184, 149 anchor RNA, 181, 105 antibodies anti-idiotypic, to opioid receptors, 178, 250 for chemiluminescent immunoassays, 184, 486 for immunoselective cell separation, 184, 316 after purification, 184, 146 via sugar moieties, 184, 162 antigens, 184, 473 bovine serum albumin, 184, 227 calmodulin, 184, 454 choleragen, 184, 405 DNA by direct incorporation of biotin, 184, 608 after modification, 184, 603 Z form, 184, 331

29 enzymatic C-terminal, proteins, 184, 160 enzymes, 184, 474 fluorescent microspheres, 184, 354 /3-galactosidase, 184, 154 glycan derivatives, 184, 654 glycoconjugates on blots, 184, 423 prior to blotting, 184, 420 glycoproteins, 184, 156 gonadotropin-releasing hormone, 184, 286 hemolytically active complement C3, 184, 621 horseradish peroxidase, 184, 150 immunoglobulins, 154, 536 insulin, 184, 247 leukocyte surface proteins, 184, 430 monoclonal antibodies for double fluorescence immunolabeling studies, 184, 346 in hybridoma culture supernatant, 184, 472 myosin ATPase site, 184, 412 nucleic acid hybridization probes, 184, 582 oligodeoxyribonucleotide probes for visualization of neuroendocrine mRNAs, 168, 753 parathyroid hormone, 184, 277 peptide hormones for flow cytometry, 184, 275 polyclonal antibodies, 184, 471 polysaccharides, 184, 539 pre-mRNA, 184, 322 protein A, 184, 498 proteins via amino groups, 184, 139 with biotinyl N-hydroxysuccinimide ester, 184, 140 on blots, 184, 422 before blotting, 184, 418 via carboxyl groups, 184, 154 with p-diazobenzoylbiocytin, 184, 151 on exocytic transport vesicle or plasma membrane surfaces, 191, 822 on nitrocellulose replicas, 184, 434

Blastema cells thiol-containing, by SH-specific reagents, 1114, 153 for visualization of intracellular trafficking, 184, 380 in purification and characterization of membrane proteins, 184, 632 transforming growth factor-B, 18,1, 279 and vectorial crosslinking, purified proteins, 184, 159 whole antiserum, 184, 148 Biotinylestradiol conjugates, preparation, 184, 292 in purification of estrogen receptor, 184. 292 synthesis, 184, 294 Biotinylglycan -avidin neoglycoproteins applications, 184, 658 preparation and properties, 184, 656 Biotinyl-N-hydroxysuccinimide ester preparation, 184, 126 Biotinyl-p-nitrophenyl ester preparation, 184, 126 Biotinylpropranolol derivatives, preparation and characterization, 184, 660 2',7'-Bis(carboxyethylcarboxy) fluorescein bleaching and leakage from cells, 192, 53 loading of parietal cells, 192, 50 in measurement of pH in parietal cells. 192, 49 Biscarboxyethyl-5(6)-carboxyfluorescein entrapment by mammalian mitochondrial matrix, 174, 115 Biscarboxyethyl fluorescein in measurement of pH gradients in cells and vesicles, 191, 472 Bismuth staining of nucleolus. 194, 723 Bladder toad, membrane vesicles, amilorideblocked Na + uptake assay, 172, 159 kinetics, 172, 161 urinary, s e e Urinary bladder Blastema cells affinity along regenerating limb axis, position-dependent differences, 190, 198

Bleaching Bleaching 2',7'-bis(carboxyethylcarboxy) fluorescein, 192, 53 photo-, see Photobleaching Bleomycin in assay of iron complexes for radical reactions, 186, 36 Blindness night, see Night blindness Bloch equations in validation of protein solution structures obtained from NMR data, 177, 245 Blood, see also Plasma, Serum arteriolar, renal collection, 191, 121 resistances, 191, 125 chicken, isolation of erythrocyte nuclei, 170, 434 citrated, preparation, 169, 51 collection for thrombospondin radioimmunoassay, 169, 262 flowing, platelet-vessel wall interactions in, measurement, 169, 37 flow rate, control, 169, 58 human, granulocyte and lymphocyte collection, 186, 228 laboratory animal, platelet isolation, 169, 29 nonanticoagulated, preparation, 169, 50 peptide transport from brain to, assay, 168, 652 porcine, platelet isolation, 169, 22 preparations for platelet aggregation studies in vitro, anticoagulant effects, 169, 147 rabbit circulating prostacylcin, assay, 169, 462 transfused, cyclic [3H]AMP, assay, 169, 468 reconstituted, preparation, 169, 52 retinoids, HPLC, 189, 50 vitamin A levels, static measures, 190, 244 Blood vessels wall components, interaction with platelets in flowing blood, measurement, 169, 37

30 Blots, see also specific blots in assay of biotin binding to streptavidin, 184, 85 proteins and glycoproteins on, analysis, 184, 415 staining, 184, 425 Blotting, see also specific techniques chromosomes, in positional mapping of yeast genes, 194, 57 direct, antibodies for immunodetection, 178, 60 DNA fragments to DBM paper, 170, 334 histones onto glass fiber membranes, 170, 475 slot, in assay of interphotoreceptor retinoid-binding protein, 189, 221 Bombesin -gastrin-releasing peptide receptors, in human and murine cells binding studies, 168, 485 characterization, 168, 481 glycoprotein analysis, 168,487 stimulation of phosphatidylinositol turnover, 168, 490 growth factor function, 168,492 internalization by Swiss 3T3 cells, 168, 489 radiolabeling, 168, 484 Bone fetal, organ cultures, 190, 423 Bone marrow derived mast cells activated, lipid metabolite production, 187, 517 cell culture, 187, 515 exocytosis of lipid mediators, 187, 516 phosphatidylinositol turnover, 187, 517 stimulated, 5-1ipoxygenase activation and exocytosis, differentiation, 187, 518 derived mesangial cells, isolation and culture, 191, 147 Borane reduction of glycolipids, 193, 755 Borax -boric acid buffer stock solutions, 182, 36 -NaOH buffer stock solutions, 182, 37 Boric acid -borax buffer stock solutions, 182, 36

31 Borohydride tritiated, in assay of carbonyl content of oxidatively modified proteins, 186, 467 Botrocetin preparation from snake venom, 169, 152 in testing for von Wiltebrand factor using fixed platelets assay of anti-vWF antibody triers, 169, 161 bioassay, 169, 155 screening for vWF multimer size, 169, 160 Bradford assay for proteins, overview, 182, 62 Bradykinin phosphorylated, tandem MS, 193, 494 Brain, see also specficic brain regions gerbil, ginkgolide interactions with platelet-activating factor binding sites, 187, 436, 440 human, hypothalamic tissue, perifusion in vitro, 168, 206 ischemia-reoxygenation injury, 186, 80 murine, carrier-mediated transport of peptides to blood, assay, 168, 652 porcine isolation and purification of angiotensin II, 168, 550 purification of glycolipid transfer protein, 179, 559 purification of cholecystokinin, 168, 303 rat cell culture angiotensin biosynthesis, 168, 555 angiotensin degradation, 168, 561 conditions for, 184, 406 detection of preproenkephalin mRNA transcripts, 168, 688 hypothalamohypophyseal axis functional analysis, microsurgical techniques, 168, 234 structural and anatomical features, 168, 234 localization of ganglioside GMt, 184, 405 median eminence and superior cervical ganglion, tissue preparation, 168, 378

BS 3 membrane opioid receptor binding studies, 178, 251,256 [3H]neurotensin-binding sites characteristics, 168,465 distribution, 168, 470 protein kinase C subspecies, resolution, 168, 347 supraoptic nucleus, hemorrhageinduced noradrenaline and 5-HJ release, assay, 168, 201 tissue microdissection, 168, 400 preparation for receptor immunocytochemistry, 178, 215 slices, preparation for assay of opioid peptides, 168, 116 tissue angiotensin, chromatographic characterization, 168, 545 Breast carcinogenesis, chemoprevention by retinoids, 190, 402 tumor induction, 190, 399 Brevibacterium fuscum

purification of 4-hydroxyphenylacetate 3-hydroxylase, 188, 112 protocatechuate 3,4-dioxygenase, 188, 85 Brij 58 prevention of loss of rRNA terminal maturase activity in solution, 181, 370 1-Bromo-2-oxo-4-(S-2,4-dinitrophenyl)mercaptobutane affinity labels for catalytic antibodies, 178, 558 l-Bromo-2-oxo-6-(S-2.4-dinitrophenyl)mercaptohexane affinity labels for catalytic antibodies, 178, 560 l-Bromo-2-oxo-5-(S-2,4-dinitrophenyl)mercaptopentane affinity labels for catalytic antibodies, 178, 559 Bryophytes general characteristics, 174, 379 transport systems, overview, 174, 366 BS 3 membrane-impermeant crosslinking reagent, 172. 619

Buds

32

Buds yeast, scars, staining with Calcofluor, 194, 732 Buffers broad-range, overview, 182, 30 for cell permeabilization studies, ion selection, 192,297 for chemical crosslinking of histones, 170, 553 for chromatofocusing proteins in different pH ranges, 182, 384 for glutaminylpeptide cyclase assay, 168, 361 for plant tissue homogenization, 182, 179 for preparation of extracts from higher eukaryotes, 182, 196 preparation, overview, 182, 29 for protein cleavage, 182, 620 for protein purification, additions to, 182, 11 for reversed-phase HPLC, 193, 404 selection, overview, 182, 26 simple, detection of catalytic metals, 186, 125 for solubilization of membrane proteins, 182, 257 stock solutions, 182, 31 theory, 182, 24 volatile, overview, 182, 29 for yeast extract preparation, 182, 167 Bumetanide 3H-labeled binding studies in intact cells, 191, 792 in identification and purification of Na+/K+/CI - cotransport system, 191, 786 a-Bungarotoxin sequential resonance assignments, 177, 141

tert-Butyldimethylsilylation for mass spectrometry, 193, 327

C Cable analysis in determination of short-circuit current in small tubular structures, 192, 15 Cacodylate buffer stock solutions, 182, 34

Calcium A23187-induced permeabilization to, 173, 107, 110 ct-adrenergic agonist-induced movement, 174, 119 binding proteins in platelet membrane, identification and characterization, 169, 365 buffering by Fura-2, 192, 72 Ca2+-2Na ÷ exchange, analysis, 174, 81 cytoplasmic, assay in living cells, 173, 745 in platelets with aequorin, 169, 387 fluorescent indicators, 169, 399 cytosolic, assay in atrial and ventricular myocytes, 173, 674 with quin2, 172, 230 in Swiss 3T3 and small cell lung carcinoma cells, 168, 490 effects on mammalian intramitochondrial dehydrogenases, 174, 95 extracellular, interaction with human platelet surface membrane, 169, 355 fluorescent indicators, entrapment by mitochondrial matrix, 174, 115 fluxes in hepatocytes, assay, 173, 534 induced Ca 2÷ release in mitochondria, analysis, 174, 79 intracellular in anterior pituitary cells assay by single-cell methods, 168, 263 spatial maps, 168, 278 effect on membrane current in intestinal secretory cells, 192, 316 free, fluorescence assay, 191, 692 in intact red cells, assay and control, 173, 100 mobilization, activation by arachidonate, 191, 684 pancreatic acinar cell levels oscillations, 192, 304 stimulant-evoked increase, 192, 301 in parietal cells, assay, 192, 60 role in basolateral K + conductance in turtle colon, 192, 742 in single isolated cells, assay, 192, 317

33 intramitochondrial enzyme Ca>-sensitive properties as indicators, 174, 107 hormone effects, 174, 111 role in regulation of oxidative metabolism, 174, 114 loading of chelator-containing red cells, 173, 109 messenger system and cAMP messenger system integration in cellular response, 191, 667 interrelationships, 191, 666 model of epithelial cell stimulussecretion coupling, 191, 664 metabolism in parietal cells, analysis, 192, 67 Na+/Ca + exchange in cardiac sarcolemmal vesicles, charge movements during, 172, 155 red blood cells, analysis, 173, 292 platelet surface-bound assay on stimulated platelets, 169, 365 unstimulated platelets, 169, 356 role in platelet function, 169, 369 pumping across plasma membrane and internal store refilling, 192, 69 in rapidly frozen cells, electron probe Xray microanalysis, 172, 203 regulated photoproteins, application as Ca > indicators, 172, 164 role in intracellular signaling, 192, 72 sensitive intramitochondrial enzymes, properties, 174, 95 sensitive probes, overview, 168, 271 shocked Escherichia coli, preparation and transformation, 194. 326 signaling in Fura-2-1oaded cells, regulating mechanisms, 191, 701 receptor-mediated, analysis, 191, 691 tolerant atrial and ventricular myocytes, isolation from adult rat, 173. 662 transport assay, 172, 41 in human red cells, 173, 101 in intact platelets, analysis with 45Ca exchange methods, 169, 371

Calmodulin by intestinal epithelial cell basolateral membrane, 192, 448 in mitochondria, 174, 68 across mitochondrial inner membrane, 174, 107 transported in synaptosomes, assay, 174, 3 transporting cells, Ca 2- as messenger in, 191,674 unidirectional fluxes in liver, assay. 192, 495 uptake during parietal cell reloading, 192, 70 Calcium channels activation by arachidonate, 191, 683 voltage-gated, characterization, 191. 745 Calcium ionophore and diacylglycerol, synergistic effects on platelet secretion, 169, 436 Calcium phosphate and DNA, coprecipitation, in DNA introduction into mammalian cells, 185, 528 mediated cotransfection, in cotransformarion of mammalian cells, 185, 563 Calcium pump, see ATPase, Ca > Calcofluor staining of yeast bud scars and cell wall chitin, 194, 732 CALCULATING LP program calculation of osmotic permeability coefficient for water, 172, 277 Calf thymus ribonuclease H in analysis of RNA-protecting factor interactions, 180, 465 digestion of genomic R N A - c D N A hybrids, in intron mapping, 180, 325 oligonucleotide-targeted, inactivation of U7 snRNPs, 181, 85 in site-specific degradation of RNA of snRNP particles, 181, 284 test substrates, 181, 190 Californium chemical properties, 193, 275 fission fragments, properties, 193, 276 nuclear decay properties, 193, 275 radiological aspects, 193, 277 Calmodulin binding peptides, proteolytic mapping, 184. 465

Calmodulin binding proteins expression, regional and subcellular differences, detection, 184, 462 identification, 184, 451 biotinylated derivatives applications, 184, 462 characterization, 184, 456 preparation, 184, 454 Calorimetry in analysis of RNA melting, 180, 323 Calpain types I and II assay, 169, 444 natural substrates, 169, 454 properties, 169, 449 purification from human platelets, 169, 445 Calpastatin assay, 169, 444 properties, 169,450 purification, 169, 445 Cancer chemoprevention by retinoids, animal models, 190, 395 Candida C.boidinii, purification of

amine oxidases, 188, 429 catalase, 188, 465 dihydroxyacetone synthase, 188, 441 formaldehyde dehydrogenase, 188, 456 triokinase, 188, 448 C. lipolytica

cell-free extracts, preparation, 188, 172, 177 subceUular fractionation, 188, 173, 177 C. methylica, purification of glycerone kinase, 188, 452 C. tropicalis

cell-free extracts, preparation, 188, 172, 177 subcellular fractionation, 188, 173, 177 C. utilis, purification of transaldolase, 182, 788 Canine parvovirus monoclonal anti-idiotypic antibody vaccines, 178, 375 Capacitance analytical determination from transmucosal potential difference, 171, 611

34 transepithelial, in mammalian urinary bladder, measurement, 192, 644 Capillaries peritubular network, permselectivity, multiple indicator dilution studies, 191, 55 permeability, as index of microvascular injury, 186, 736 Capillary columns for GC/MS of permethylated oligosaccharides, preparation, 193, 735 Capillary electrophoresis with continuous-flow FAB/MS, peptides and proteins, 193, 233 with electrospray ionization MS, 193, 430 Capnophorin, see Proteins, band 3 Carbachol dose-response testing by hippocampal brain slice recording, 168, 140 effect on prostaglandin E2 production by parietal cells, 187, 513 Carbacholine effect on gastric glands, 191, 728 Carbamoyl-phosphate synthase -aspartate transcarbamoylase-dihydroorotase, in gene selection and coamplification, 185, 550 Carbenes general properties, 172, 635 and precursors, properties, 172, 641 Carbocyanines in determination of membrane potential in murine ascites tumor cells, 172, 95 optical probes for, 172, 76 in yeast, 174, 615 Carbodiimide, see Cyanamide Carbohydrate receptors binding activities agglutination assay with gold complexes, 179, 120 detection by colloidal gold, 179, 111 microscopic evaluation, 179, 117 Carbohydrates asparagine-linked, attachment site and structural class in glycoproteins, 193, 501 binding by snowdrop lectin, specificity, 179, 329

35 biological, analysis, 193, 546 charged and electroneutral, accumulation by H + cotransport in Rhodotorula, 174, 629 complex, metabolic labeling with radiolabeled hexosamines, 179, 435 derivatized surfaces for cell growth, 179, 555 in glycolipids, nomenclature, 193, 741 in glycoproteins, radiolabeling, 194, 690 in glycosphingolipids, sequence information from MS analysis, 193, 632 in human tracheobronchial mucin glycoproteins, analysis, 179, 14 immobilization on polyacrylamide gels, 179, 542 immobilized in microtiter wells, cell adhesion to, 179, 547 linkage analysis by Lindberg method, 193, 554 mass calculations, 193, 515 methylated, quantitative hydrolysis, 193, 559 methylation, 193, 556 permethylated, FAB/MS, 193, 518 in protein, addition or processing in mammalian cells, inhibitors, 185, 589 specific human monoclonal antibodies, production in vitro, 179, 299 transport, catalyzing carriers, number in Rhodotorula, 174, 638 Carbon assignment in heteronuclear NMR, 176, 108 ~3C, spin-lattice relaxation, 176, 188 -carbon correlations in 2D NMR of proteins, 176, 41 films coating of electron microscope grid, 170, 167 in immunoelectron microscopy of idiotype-anti-Id complexes, 178, 133 support on grid for chromatin deposition, preparation, 180, 483 surface properties, modification, 170, 171

Carboxylic acids grid coating for electron microscopy of chromatin, 170, 143 -nitrogen correlations in 2D NMR of proteins, 176, 46 Carbonate -bicarbonate buffer stock solutions, 182. 37 -Fe(IV) complex, generation in aqueous solutions, 186, 111 Carbonate dehydratase I Mn(II)-, chemical exchange reactions, magnetization transfer NMR study. 176, 320 Carbon dioxide -bicarbonate solutions, pH control. 192. 42 dissolved in yeast, continuous estimation, 174, 602 induced exocytotic H*-ATPase insertion, in regulation of urinary epithelial H + transport, 172, 54 production by parietal cells, assay, 192. 104 Carbonyl groups in oxidized proteins, assay, 186, 464 triplet, chemiluminescent, chemistry and sources, 186, 596 Carboxy-2',7'-dichlorofluorescein diacetate in vital staining of yeast vacuoles, 194. 646 Carboxyftuorescein permeability of phospholipid membranes, effects of retinoids. 189, 379 6-Carboxyfluorescein acetate indicator for analysis of transport in isolated renal cells, 191,404 6-Carboxyfluorescein diacetate in vital staining of yeast vacuoles, 194. 646 Carboxylation vitamin K-dependent protein, role of DT-diaphorase, 186, 297 /3-Carboxylation aspartate residues, 182, 635 7-Carboxylation glutamate residues, 182, 634 Carboxylic acids labeling by exchange with H?80, 193, 342

Carboxyl oxygen exchange Carboxyl oxygen exchange in synthesis of [170] or [180]carboxylenriched homoprotocatechuate, 188, 110 Carboxymethylation glycoproteins, 193, 508 proteins, 182, 599 Carboxypeptidase S yeast biochemical analysis, 185, 385 enzymatic assay, 194, 449 genetic analysis, 185, 377 well test, 194, 439 Carboxypeptidase Y, see Serine carboxypeptidase Carcinogenesis breast, chemoprevention by retinoids, 190, 402 Carcinogens binding to genome, mapping, 170, 249 a-Carotene and other nutrients in serum and plasma, simultaneous HPLC, 189, 155 /3-Carotene cleavage, assay and biological significance, 189, 434 incorporation into mixed micelles, 189, 418 isomers, reversed-phase separation, 189, 175 and other nutrients in serum and plasma, simultaneous HPLC, 189, 155 and retinol, retinoid synthesis from, assay and characteristics, 189, 470 serum, liquid chromatographic assay, 189, 172 /3-Carotenoid 15,15'-dioxygenase assay, 189,426 properties, 189, 431 Carotenoids analysis, 190, 238 cleavage pathways, 189, 433 conversion reactions, 189, 425 excentric cleavage, 189, 431 supplying vitamin A and provitamin A, food composition tables for, assessment, 190, 237 Carrier molecules carbohydrate transport-catalyzing, number in Rhodotorula, 174, 638

36 implantation into cells by Sendai virus envelope-mediated fusion, 171, 829 inhibition kinetics, 171, 113 kinetic comparison with pores, 173, 152 mediated transport bile acids, 192, 531 ions through membranes, kinetics, 171, 62 steady-state rate equations, generation, 171, 164 simple, kinetics, 173, 149 Cartilage pattern in developed embryo, analysis, 190, 206 pieces, culture, 190, 426 Caseinolytic assay calpains I and II, 169, 444 CAT, see Chloramphenicol acetyltransferase Catalase assay, 188,463 effects on endotoxin-induced circulatory shock, 186, 661 endotoxin-induced disseminated intravascular coagulation, 186, 657 localization in Hansenula polymorpha, 188, 413 peroxisomal marker, assay, 182, 219 properties, 188, 466 purification from Candida boidinii, 188, 465 role in myocardial preservation during heart transplantation, 186, 742 Catecholamine receptors /3-adrenergic, anti-idiotypic antibodies production and characterization, 178, 272 responses in mouse, 178, 272 rabbit, 178, 269 Catecholamines plasma HPLC, 168, 439 radioenzymatic assay, 168, 435 Catechol 1,2-dioxygenase assay, 188, 123 purification from Acinetobacter calcoaceticus, 188, 124

37 yield, storage, stability, and properties, 188, 126 Catechol 2,3-dioxygenase amino acid composition, 188, 119 assay, 188, 116 induction, 188, 120 physiological role, 188, 120 properties, 188, 118 purification from Pseudomonas aeruginosa, 188, 116 Catechol estrogens o-semiquinones from, spin stabilization, 186, 341 Cations alkali metal, transport in Rhodotorula, 174, 645 and anions, cotransport electrogenicity and electroneutrality. 173, 287 stoichiometry, 173, 283 in buffers for cell permeabilization studies, selection, 192, 298 composition in yeast cells, alteration, 174, 607 divalent brush border membranes free of. isolation, 192, 353 precipitation of brush border membranes, 192, 353 fluxes in red blood cells, 173, 80 intracellular content in erythrocytes, alteration, 173, 81 monovalent, binding by yeast, 174, 610 organic, transport across reptilian proximal tubules, mechanism, 191, 218 sulfur-centered radical, chemical properties, 186, 178 tandem mass spectrometry calibration mixture, 193, 873 uptake by yeast, effect of cell pH, 174, 608 Cavitation disruption of cells, 172, 6 Cell adhesion effects of retinoids, 190, 81 Cell blot assay hormone secretion from single cells, 168, 327 Cell counting on chemotaxis membranes, 169. 241

Cell cultures Cell cracking for permeabilization to macromolecular probes, 168, 225 Cell cultures Acetabularia, 174, 490 anterior pituitary cells, 168, 146 BALB/c 3T3 cells for assays of plateletderived growth factor, 169, 214 /3 cell lines, 192, 223 bone marrow-derived mast cells, 187, 515 bovine corneal endothelial cells, 192. 571 cardiac cells from chicken embryos and rat neonates, 173, 637 cell type enrichment methods, 171, 492 dishes, coating with antibody for tissue immunodissection, 171, 585 embryonal carcinoma cells, 189, 338 epithelial, porous-bottom dishes for, in transport and differentiation studies, 171, 736 fibroblasts, 190, 176 glomerular endothelial cells, 191, 148 glomerular epithelial cells, 191, 150 glomeruli, 191, 141 HeLa cells labeling, 180, 413; 181, 321 preparation, 181, 6, 240 intracellular environment, experimental control, 171, 817 juxtoglomerular interstitial cells, 191, 152 keratinocytes air-exposed system, 190, 31 culturing at air-liquid interface, 190, 23 submerged systems, 190, 20, 31 limb bud, for tetatogenicity studies, 190, 422,427 mammalian cells optimization for recombinant protein production, 185, 567 for preparation of active nuclei, 181, 32 MDCK cells, 191, 815 media for, see Culture media mesangial cells, 191, 146 Methylobacterium extorquens AM1, 188, 374

Cell c u l t u r e s monolayer cell release by trypsin and EDTA, 171, 567 hepatocytes amino acid transport assay, 173, 564 preparation, 173, 567 solute fluxes and binding, measurement devices, 171, 133 M y c o b a c t e r i u m , 188, 149 neuronal and glial cells, 168, 558 pH and proton pumping, assay, 174, 140 pituitary, for bioassay of inhibin, 168, 602 preparation of nuclei and homogenates, 170, 320 primary airway epithelial cells, 192, 566 gastric mucosa endocrine cells, 192, 181 intact islets of Langerhans and dispersed islet cells, 192, 188 isolated tubule cells of defined segmental origin cell isolation, 191,410 growth and differentiation requirements, 191,415 mesangial cells, 187, 547 rat brain, conditions for, 184, 406 rat endothelial cells, 187, 400 rat vascular smooth muscle cells, 187, 400 renomedullary interstitial cells, 191, 162 retinoblastoma, 190, 141 sebocytes, conditions for, 190, 335 Sertoli cells, preparation and effects of retinoids, 19tl, 71 static, for prolactin secretion assay, 168, 254 subculturing, T84 cells, 192, 388 suspensions, s e e Suspension cultures three-dimensional, anterior pituitary cells, 168, 48 tracheal epithelial cells, 192, 560 transport and metabolism measurements, overview, 173,714 yeast growth, 194, 12 growth conditions after mutagen treatment, 194, 274

38 Cell disruption by cavitation, 172, 6 effect on multienzyme complex purification, 182, 541 by shearing forces, 172, 5 Cell fusion facilitated, for hybridoma production, 184, 641 MDCK cells, 191, 439 peripheral lymphocytes in vitro, 182, 677 renal epithelial cells, 192, 715 spleen cells with myeloma cells, 182, 675 Cell injury correlation with free radical formation, spin trapping studies, 186, 611 Cell lines A-431 (human epidermis), glycolipid purification, 179, 43 ascitic, preparation, 171, 593 CHO (Chinese hamster ovary), s e e CHO cells COS-1 (African green monkey kidney), transient expression of plasmid DNA, 185, 507 CV1 (African green monkey kidney) cytosol preparation, 170, 47 infection with simian virus 40, 170, 15, 46 nuclear extract preparation, 170, 46 eukaryotic, CAT assay interference, 168, 698 heavy-chain loss variant, for production of chimeric antibody, selection, 178, 522 HeLa, s e e HeLa cells intestinal availability, 192, 355 Cl- channels, analysis, 192, 377 electrolyte transport study models, 192, 354 K ÷ channels, analysis, 192, 372 Na÷/H ÷ exchange and other Na + transport pathways, analysis, 192, 381 Na+/K+/CI- cotransport pathway, analysis, 192, 371 radionuclide uptake and efflux studies, 192, 367 T~

39 growth and maintenance, 192, 357 morphological studies, 192, 359 trypsinization, subculturing, and plating, 192, 388 transepithelial electrolyte transport, analysis, 192, 361 mammalian constitutive expression of proteins. 182, 115 isolation of subcellular organelles, 182, 203 nucleus, retinoic acid-binding protein and receptor identification and analysis, 189, 239 for production of recombinant proteins, selection, 185, 567 retinoid synthesis from retinol and/3carotene, assay and characteristics, 189, 470 MDCK (Madin-Darby canine kidney) giant cell fusion, 191,439 microinjection of mRNA, 191, 443 preparation for microinjection, 191, 439 signal analysis, 191, 448 polarized, recovery of exocytic transport vesicles, 191, 813 renal bidirectional transepithelial solute fluxes, 191,436 epithelial, phosphate transport, 191. 494 mycoplasma contamination, detection, 191,430 tissue culture, 191,426 transepithelial electrical properties, measurement, 191, 435 transmembrane solute transport, measurements, 191,430 retinoid-sensitive, synopsis, 190, 217 Sf9 (Spodoptera frugiperda) culture media, growth, and transfection, 182, 121 -lytic baculovirus system for mammalian protein expression, 182, 117 Cell populations separation by free-flow electrophoresis, 171. 513

Central nervous s y s t e m Cell proliferation epidermal, retinoid effects, 190, 77 regulation by gangliosides, 179, 521 Cells retinoid-sensitive, synopsis, 190, 217 Cell sorters fluorescence-activated, see Fluorescence-activated cell sorter Cell sorting cell preparation and labeling for, 171. 572 cell viability after, enhancement, 171. 57O electrostatic, process components. 171. 557 speed, relationship to purity and percentage recovery, 171, 571 sterile techniques. 171, 569 Cell swelling Na+/H + exchange-induced, detection, 173, 779 Cellulose -biotin, for affinity chromatography, synthesis, 170, 49 Cellulose acetate in fractionation of RNA digests, 180. 137 Cell viability after cell sorting, enhancement, 171, 570 glomerular mesangial cell cultures, 187. 552 testing by plasmolysis, 174, 246 Cell volume epithelial, regulation, experimental approaches, 171, 744 in isolated renal tubules, measurement. 191, 371 measurement, in detection of Na*/H ~ exchange in lymphocytes, 173, 777 pressure probe measurement, constraints, 174. 356 Cell wall elastic modulus determination, 174, 353 Central nervous system neuropeptide gene transcription, assay, 168, 681 neurotensin binding specific sites, distribution, 168, 470 in tissue membranes and sections, assay. 168, 464

Central nervous system peptide activity in, effects on autonomic nervous system, 168, 432 repeffusion injury, role of oxygenderived free radicals, 186, 717 Centrifugation in assay of anti-antimorphine activity, 178, 228 buoyant density, in cell separation, 171, 497 in cesium chloride gradients DNA-protein complexes, 170, 393 isopycnic, core snRNPs, 181, 271 U l and U5 snRNPs in native form, 181, 267 versatility, 181, 270 density gradient bovine synaptic vesicles, 174, 14 corticoid receptors in glycerol, 191, 535 enterocyte subcellular fractionation, 192, 345, 350 gastric cell purification, 171, 451 H+,K+-ATPase-containing membrane isolation, 192, 154 leaf material, 174, 537 nonparenchymal liver cells in Percoll gradients, 190, 66 plasma membrane preparation, 172, 9 red outer medulla lumenal membrane preparation, 191,463 renal cortical basolateral plasma membrane purification, 191, 456 subcellular membrane isolation, 172, 2O two-stage, in preparative-scale transport-specific fractionation, 172, 48 differential exocytic transport vesicle isolation, 191, 817 iterative, gastric cell purification, 171, 452 plasma membrane preparation, 172, 9 renal cell lumenal plasma membrane enrichment, 191, 461 total liver cell suspension, 190, 65 in formation of transporting large proteoliposomes, 191, 595 glomeruli, 191, 133 gradient, for cell separation, 171, 462 human platelet isolation from plasma, 169, 3

40 hypaque gradient, renal basolateral membrane vesicle purification, 191, 464 in liposome particle size determination, 171, 216 micro-, s e e Microcentrifugation in orientation of synthetic and native membranes, 172, 401 PercoU isoneutral, gastric cell purification, 171, 452 in platelet washing to obtain platelet yon Willebrand factor, 169,246 in protein concentration and solute removal, 182, 83 protoplasts in silicone oil, 174, 520 sucrose gradient, data analysis methods, 172, 707 tracheobronchial mucin, 179, 15 ultra-, s e e Ultracentrifugation yeast vacuole purification, 174, 508 Centromeres yeast, linkage, and single gene segregation, 194, 41 Ceramide glycanase assay, 179,479 isolation from earthworm, 179, 482 leech, 179, 481 properties, 179, 482, 484 Ceramides in glycosphingolipids, composition, 193, 637 portion of glycolipids, nomenclature, 193, 741 positive ion tandem mass spectrometry, 193, 744 reversed-phase HPLC, 172, 556 Cesium chloride in fractionation of yeast DNA, 194, 174 in purification and characterization of U snRNPs, 181, 264 Cesium iodide ion reference masses, 193, 874 and NaI and RbI, positive ion mixture, reference masses, 193, 873 Cetavlon, s e e Cetrimonium bromide Cetrimonium bromide in depletion of uncrosslinked proteins from DNA-protein sample, 170, 393 precipitation of mannoprotein, 185,456

41

Chernotaxis

Cetylpyridinium chloride induced precipitation, in platelet heparitinase assays, 169, 343 Chaotropic agents in buffers for solubilization of membrane proteins, 182, 258 in RNA preparation, 180, 3 Chara adenine nucleotide levels, effect on ion transport, 174, 472 cytoplasmic pH, effect on ion transport, 174, 473 internodal cells, isolation by cell surgery, 174, 444 intracellular perfusion, 174, 446 ion transport studies, 174, 443 plasma membrane isotopic fluxes, 174, 450 net fluxes, 174, 459 tonoplast, ion fluxes, 174, 455 transport kinetics, 174, 475 vacuolar contents, sampling, 174, 445 Charcoal dextran-coated, in cellular retinoidbinding assays, 190, 152 immunoassay with, secretin, 182, 711 Checkerboard analysis in confirmation of chemotactic activity, 169, 241 Chelation therapy site-specific antioxidant protection, 186, 41 Chemical exchange influenced NMR line shapes, computation, 176, 282 magnetization transfer NMR studies, 176, 311 quadrupolar relaxation in absence of, 177, 319 Chemical shifts dependence on molecular conformation, 176, 368 fluorine, data interpretation, 177, 16 due to gramicidin channel interactions, 171, 324 ion resonance, in ion binding constant determination, 171, 314 31p

bond-angle effects, 177, 300 extrinsic effects, 177, 306 principles, 177, 298

stereoelectronic effects description, 177, 302 as probe of DNA and DNA-protein structures, 177, 310 theoretical calculations and empirical observations, 177, 299 phosphoproteins in 3~p NMR, 177, 268 quadrupolar nuclei in ligand-protein complexes, 177, 319 site (carbon-13), in ion binding constant determination, 171, 316 Chemiluminescence in assay of nitric oxide, 186, 610 enzyme-dependent, singlet molecular oxygen, 186, 135 generating species, chemistry and sources, 186, 596 in HPLC assay of lipid hydroperoxides. 186, 371 luciferin analogs, in assay of superoxide dismutase, 186, 227 superoxide radicals and singlet oxygen, 186, 585 visible-range, low-level, in biological systems antioxidant effects, 186, 608 detection 186, 597 emission enhancers, 186, 601 quenchers, 186, 603 relationship to lipid peroxidation parameters, 186, 609 spectral analysis, 186, 600 two-dimensional imaging, 186, 604 Chemiosmosis nonequilibrium thermodynamics, 171, 439 Chemoprevention breast carcinogenesis by retinoids, 190, 402 cancer by retinoids, animal models, 190, 395 Chemostat in preparation of yeast with high permease activity, 174, 628 Chemotaxis assays, membranes for, cell counting on. 169, 241 inflammatory cells and fibroblasts induced by platelet-derived growth factor assays, 169, 238

Chemotaxis checkerboard analysis, 169, 241 Chirality eicosanoids analysis by reaction with lipoxygenase, 187, 194 HPLC analysis, 187, 187 nomenclature, 187, 187 Chitin binding for isolation of chitinase secreted by yeast, 194, 696 in yeast cell wall, staining with Calcofluor, 194, 732 Chitinase immunoprecipitation, 194, 694 O-linked mannooligosaccharides from, analysis, 194, 691 O-mannosylated, mobility shifts, 194, 687 secreted, isolation by chitin binding, 194, 696 Chloral hydrate anesthetic for in vivo analysis of opioid peptides, 168, 104 Chloramine-T in direct iodination of proteins, 182, 723 Chloramphenicol acetyltransferase assay, 189, 269 reporter gene, assay, 168, 690 interferences in eukaryotic cell lines, 168, 698 interferences from hepatoma cell extracts abolition by EDTA and heat, 168, 695 identification, 168,693 Chlorate containing medium, cells incubated in, undersulfation of proteoglycans, 179, 429 Chlorella

amino acid and sugar transport efflux and influx measurements, 174, 392 energization, 174, 396 inducibility and specificity, 174, 391 C. kessleri, amino acid and sugar uptake systems, inducibility and specificity, 174, 391 transport mutants, isolation, 174, 398 transport protein identification, 174, 399

42 Chloride -alkali metal cotransport, analysis, 173, 345 -bicarbonate exchange, detection and analysis, 173, 344 Ca2+-dependent current, in analysis of [Ca2+]i oscillations, 192, 304 cellular leak in turtle colon, 192, 737 CI /C1- exchange in intestinal membrane vesicles, 192, 408 36C1- influx, effects of furosemide-related diuretics, 191, 790 coulometric titration, 173, 62 effects on phenylglyoxal modification of anion transport system, 173, 473 exchange, reversible inhibition by phenylglyoxal, 173, 476 flux, as function of external concentration, 173, 441 loading of erythrocytes, 173, 438 Na+/K+/C1 - cotransport in intestinal cell lines, 192, 371 system identification and purification, 191,786 radioisotopic erythrocytes with, preparation, 173, 58 flux experiments, 173, 63 role in amino acid transport in red cells, 173, 146 secretion in shark proximal tubules, 191, 184 stimulation of vesicular glutamate uptake, 174, 20 transport in avian renal tubules, 191, 226 intestinal membrane vesicles, 192, 405 kidney, effects of furosemide-related diuretics, 191, 785 nutrient membrane, mechanistic analysis, 171,622 uptake by teleost intestine, initial rates, 192, 750 voltage-sensitive fluxes, prediction and verification, 174, 501 Chloride channels blockers acting in apolar cells, 191, 801 epithelia, 191, 802 muscles, 191, 800 neurons, 191,799

43 epithelial purification and reconstitution, 191, 572 solubilization, 191, 575 general properties, 191,794 in intestinal cell lines, 192, 377 in turtle colon basolateral membrane, 192, 741 Chloride peroxidase catalyzed ethyl hydroperoxide decomposition, tO2 detection during, 186, 133 Chloride pump ATP-driven, in Acetabularia electrical properties, 174, 493 electrogenic, demonstration, 174, 492 kinetic models, 174, 498 reversal in vivo, ATP synthesis, 174, 501 Chlorocatechol 1,2-dioxygenase assay, 188, 123 purification from P s e u d o m o n a s B 13, 188, 125 yield, storage, stability, and properties, 188, 126 Chloroform and phenol, extraction of DNA-protein complexes, 170, 394 p-(Chloromercuri)benzoic acid-Sepharose synthesis, 173, 503 Chloroplasts extract preparation, 182, 186 H+-ATPase, coupling rules, 171, 158 N-Chlorosuccinimide digestion of proteins for mass spectrometry, 193, 372 CHO cells intracellular glutathione, assay, 186, 706 subcellular organelles, isolation, 182, 203 survival after heat and thiol treatment, assay, 186, 701 thermosensitivity, thiol effects, 186, 696 thermotolerance, 186, 700 thiol-promoted cystine uptake, assay, 186, 707 Cholecystokinin CCK-8, tandem MS study, 193, 499 precursor, see Procholescystokinin purification from brain, 168, 302 Cholecystokinin octapeptide mediated channel opening, demonstra-

Chromatin tion by patch-clamp methods. 171. 667 Choleglobin absorption spectra, 186, 270 Choleragen biotin labeling, 184, 405 biotinylated, in localization of ganglioside GM~,184, 405 Cholera toxin B subunit, ligand-binding domain, analysis with anti-Id antibody to GM~, 178, 165 Cholesterol and DMPC, vesicles, spontaneous insertion of membrane proteins, 171, 261 Cholesterol 5,6-epoxides chemical reduction analysis, 186, 442 formation and assay, 186, 438 IH NMR, 186, 442 HPLC, 186, 441 recovery, 186, 440 Cholesterol sulfate relative cellular content, estimation, 190, 45 Cholesterol sulfotransferase assay, 190, 48 down-regulation by retinoids. 190, 42 Chondroitin microsomal sulfation, 179, 422 properties, 179, 426 Chromanols binding to membrane suspensions, 186, 359 esterified, hydrolysis in membrane suspensions, 186, 358 incorporation into membrane hydrophobic core and cluster formation, 186. 362 topography and mobility in lipid bilayer. 186, 367 Chromatin assembly in vitro with egg nucleoplasmin, 170, 612 method selection, 170, 575 with oocyte extracts, 170,603 digestion, in analysis of nucleosome positioning, 170, 409 DNA cleavage, 170. 410

Chromatin lysine residues binding to, radiolabeling, 170, 369 Drosophila, spreading, 180, 487 electron microscopy, 170, 142 fractionation, 170, 127 fragmentation, 170, 6 functional studies with immunological probes for nucleosomes, 170, 241 histories neighbors, determination, 170, 563 oligomeric state, determination, 170, 557 pairs, purification, 182, 335 hypersensitive sites, analysis, 170, 269 immature identification, 170, 303 nucleosome structure and spacing, 170, 304 as immunogen for antihistone antibodies, preparation, 170, 255 long fragments, preparation, 170, 9 maturation, nuclease analysis, 170, 303 Miller spreading method, 180, 482 native state preservation, criteria, 170, 7 nonenzymatic cleavage, 170, 359 nuclease specificity on, 170, 292 organization at replication forks, visualization, 170, 290 poly[d(A-T)], nucleosome spacing periodicity for, analysis, 170, 600 preparation enzymatic method, 170, 127 from eukaryotic cells, 170, 3 from HeLa and Xenopus kidney cell cultures, 170, 348 psoralen crosslinking, 170, 155 purification, 170, 6 reconstitution from purified components, 170, 585 related structures, complete reconstitution, 170, 598 replicating, nuclease analysis, 170, 303 restriction endonuclease cutting and ligation, 170, 12 simian virus 40, structure at replication forks, 170, 291 soluble attachment to grids, 170, 145 DNA-protein crosslinking, 170, 389 preparation, 170, 143, 146

44 spreads, gene identification, 180, 508 structure analysis by nuclease digestion, 170, 339 relationship to DNA sequence at replication forks, 170, 312 templates allowing transcription, reconstitution, 170, 581 RNA polymerase IlI transcription, analysis in vitro, 170, 347 transcriptionally active, nuclease digestion, 170, 317 yeast plasmid, purification biochemical method, 170, 29 protein-nucleic acid affinity method, 170, 35 Chromatofocusing high-performance, comment, 182, 391 proteins, 182, 380 in denaturing agents, 182, 390 experimental design, 182, 386 mechanism, 182, 381 reagents and equipment, 182, 381 resolution, influencing factors, 182, 383 Chromatography, see also specific techniques N-acetylglucosaminyltransferase II, 179, 473 acyl-CoA synthetases, 187, 240 alcohol dehydrogenase isoenzymes in retinoid metabolism, 189, 438 alcohol oxidase, 188, 423 aldehyde dehydrogenase, 188, 329 aldehyde dehydrogenase (acceptor), 188, 323 amine oxidases, 188, 429 in analysis of peptide processing in vivo, 168, 521 anion-exchange protein from erythrocyte membrane, 173, 415 arachidonate 5-1ipoxygenase, 187, 271, 300 in assay of /3-carotene incorporated in micelles, 189, 421 cellular retinoid binding, 190, 152 lecithin-retinol acyltransferase, 189, 455

45 bands 2.1 and 4.1 from erythrocyte membranes, 173, 385 band 3 protein, 173, 501,503 cis-benzene dihydrodiol dehydrogenase, 188, 136 benzene dioxygenase components, 188, 57 benzoate-CoA ligase, 188, 157 benzoyl-diacylglyceride subclasses, 187, 209 blue copper proteins, 188, 285 branch points from nuclear RNAs, 180, 182 calpains I and II, 169,445 calpastatin, 169, 445 cap structures, 180, 168 catalase, 188,465 catechol 2,3-dioxygenase, 188, 116 cellular retinoic acid-binding proteins from human placenta, 189, 334 from neonatal rat skin, 189, 310 types I and II, 189, 357 in characterization of angiotensin in brain tissue, 168, 545 covalently bound pyrroloquinoline quinone, 188, 277 cyclohexanone 1,2-monooxygenase, 188, 74 cyclopentanone 1,2-monooxygenase, 188, 79 cytochrome P450 prostaglandin ra-hydroxylase, 187, 255 cytochromes crt and CL, 188, 305 for detergent removal from membrane proteins, 182, 279 purified cytochrome P450 prostaglandin o~-hydroxylase, 187, 259 diacylglycerides, 187, 207 dichloromethane dehalogenase, 188, 357 5(S),12(S)-dihydroxy-6,10-trans-8,14-ciseicosatetraenoic acid, 187, 305 DT-diaphorase, 186,290 electron-transfer flavoproteins, 188, 309 endo-/3-galactosidase releasing Gal(cd---~3)Gal, 179, 498 endoglycoceramidase, 179, 490 endo-cd---,6-mannanase, 185, 462 exo-cd--*2-mannosidase, 185,463 extracellular superoxide dismutase, 186, 263

Chromatography follicle-stimulating hormone-releasing protein, 168,617 formaldehyde dehydrogenase, 188, 456 formaldehyde dehydrogenase (glutathione), 188, 316 formaldehyde transketolase, 188, 441 formate dehydrogenase from Methylosinus trichosporium OB3b 188, 332 methylotrophic yeasts, 188, 460 gentisate 1,2-dioxygenase, 188, 104 glucose-6-phosphate dehydrogenase from Arthrobacter globiforrnis, 188, 341 Pseudomonas W6, 188, 348 glucose transporter from human erythrocytes, 174, 42 glucosidase I, 179, 457 glucosidase II, 179,462 glycerone kinase, 188, 452 glycolipid transfer protein, 179, 562 glycopeptidolipids from Mycobacterium aoium, 179, 221 3-hexulose-6-phosphate synthase from Acetobacter methanolicus, 188,403 Bacillus C1,188, 394 Mycobacterium gastri MB19, 188, 399 histone H1 variants, 170, 528 histones, 170, 624 hydrocarbon monooxygenase, 188, 7 5(S)-hydroperoxyeicosatetraenoic acid, 187, 304 hydroxybenzoate hydroxylase, 188, 141 hydroxypyruvate reductase, 188, 374 immunoglobulin from serum, 182, 669 inhibin, 168, 617 inositol phosphates, 173, 536 lectins from Allomyrina dichotorna, 179, 332 leukotriene A4 hydrolase, 187, 329 leukotriene C4 synthase, 187, 310 lignin peroxidase, 188, 166 lysosomal C~-D-mannosidase,179, 501 macrophage phospholipase A~., 187, 223 malyl-CoA lyase, 188, 382 c~-mannosidase, 185,465 mannosidase I, 179, 465 mannosidase IA, 179, 447 mannosidase II, 179,465 mannosyl-glycoprotein endo-/3-N-acetylglucosaminidase, 179. 509

Chromatography membrane skeletal proteins from Triton shells, 173, 389 mercaptan-specific antioxidant protein, 186, 482 methane monooxygenase, 188, 197 methanol dehydrogenase from Bacillus C1,188, 224 Hyphornicrobium X, 188, 206 Methylobacterium extorquens AM1, 188, 213 methanol dehydrogenase protein M, 188, 218 methylamine dehydrogenase from Methylobacillus flagellatum, 188, 248 methylotrophic bacteria, 188, 242 Thiobacillus uersutus, 188, 236 methylamine oxidase, 188, 229 mRNA 3'-end-processing-related factors, 181, 71 muconate cycloisomerase, 188, 128 muconolactone isomerase, 188, 132 naphthalene dioxygenase, 188, 49 neutral lipids, 191, 687 nuclear cap binding proteins, 181, 212 nuclear extracts with histone-specific RNA 3' processing activity, 181, 87 nucleoplasmin, 170, 617, 621 oligosaccharides constituents of trehalose-containing mycobacterial lipooligosaccharides, 179, 225 haptenic, from Mycobacterium avium, 179, 221 N-linked, from yeast mnn mutants, 185, 457 pepsinogen, 192, 133 peptide N4-(N-acetyl-fl-glucosaminyl) asparagine amidase, 179, 509 phenolic glycolipid I from Mycobacterium leprae-infected armadillo tissue, 179, 227 6-phosphogluconate, 188, 343 phospholipids in balance sheet analysis of phosphatidylinositol, 191, 687 derivatized molecular species, 187, 205 major classes, 187, 198 phosphomannosyl receptors, 179, 306 Pi/H + symporter, 173, 734

46 phthalate dioxygenase, 188, 63 phthalate dioxygenase reductase, 188, 65 physalaemin-related immunoreactive peptides, 168, 451 platelet-activating factor acetylhydrolase, 187, 352 platelet-derived growth factor, 169, 216, 220 platelet heparitinase, 169, 346 polyadenylate-binding protein, 181, 336 polyadenylate polymerases, 181, 163 in polyadenylate-specific fractionation on oligo(dT)-cellulose, 180, 414 polyadenylylated RNAs, 180, 79; 181, 57 propane-specific alcohol dehydrogenase, 188, 23 prostaglandin H2, 187, 245, 248 protein kinase C subspecies, 168, 348 proteins on hydroxyapatite columns, 182, 329 protocatechuate 2,3-dioxygenase, 188, 98 protocatechuate 4,5-dioxygenase, 188, 92 pyrroloquinoline quinone, 188, 269 quinohemoprotein alcohol dehydrogenase, 188, 37 in recovery of DNA-protein complexes, 170, 394 retinal, 189, 429 retinoid metabolites, 189, 533 retinol-binding proteins from plasma, 189, 195 RNA on cellulose, 180, 430 rapid column method, 180, 376 RNase III, 181, 194 RNase BN, 181, 432 RNase D, 181, 424 RNase T, 181, 428 RNA 3'-terminal phosphate cyclase, 181, 504 serine hydroxymethyltransferases, 188, 368 sialate 9-O-acetylesterase, 179, 412 small nuclear ribonucleoproteins particles active in RNA processing, 181, 220 U1 and U5, 181, 269 solubilized protein aggregates from Escherichia coli, 182, 273 soluble cytochromes c, 188, 291

47 soluble cytochromes cu and CL, 188, 301 soluble methane monooxygenase, 188, 183 spectrin, 173, 384 splicing activities from HeLa nuclear extracts, 181, 11 tracheobronchial mucin, 179, 11 transaldolase isoenzymes, 188, 407 transforming growth factor-/3, 184, 280 trehalose-containing mycobacterial lipooligosaccharides, 179, 225 trimethylamine dehydrogenase, 188, 256 triokinase, 188, 448 tRNA nucleotidyltransferase, 181, 436 tRNA splicing endonuclease, 181, 475, 516 vaccinia virus mRNA capping enzyme, 181, 177 vasopressin-converting aminopeptidase, 168, 387, 390 vegetative tissue lectins, 179, 344 wheat germ RNA ligase, 181, 485 yeast splicing extracts, 181, 95 yeast tRNA ligase, 181,468 Chromogenic substrate assay Z-Lys-SBzl, oocyte tissue plasminogen activator, 168,425 Chromophores hydrophobic, oligosaccharides coupled to, desorption MS, 193, 661 Chromosomes components, immunocytochemical studies, 170, 234 mini-, s e e Minichromosomes polytene immunocytochemical studies, 170, 236 immunofluorescence studies, 170, 241 simian virus 40 extraction, 170, 300 replicating affinity isolation, 17tl, 41 enzymatic digestion, 170, 297 incorporation of Bio-19-SS-dUTP, 170, 45 preparation and radiolabeling, 170, 299 yeast allele recovery, 194, 298

Circulatory shock blotting, in positional mapping of genes, 194, 57 footprinting by primer extension, 194. 557 fragmentation in gene mapping, 194, 51 in positional mapping of genes, 194. 57 fragments nomenclature, 194, 76 strains with, availability, 194, 76 genetic and physical maps, 194, 827 hybridization to, in gene mapping, 194, 47 localization of Ty-induced mutations. 194, 356 segregation analysis in meiosis with yeast artificial chromosomes, 194, 765 mitotic analysis with chromosome fragments, 194, 756 structural analysis with H O endonuclease, 194, 143 yeast artificial, s e e Yeast artificial chromosomes Chylomicrons vitamin A-labeled, preparation, 190, 306 Chymotrypsin cleavage of anion transporter, 173, 425 digestion of proteins for MS analysis, 193, 367 Cigarette smoke affected human lung, preparation and ESR analysis, 186,-665 Ciliary body rabbit, ginkgolide interactions with platelet-activating factor binding sites, 187, 437. 440 Cimetidine effects on gastric gland preparations, 191, 729 isolated gastric mucosa, 191. 724 Circular dichroism in determination of membrane protein secondary structure and aggregation state, 172, 575 solubilized band 3 protein. 173, 511 Circulatory shock endotoxin-induced, role of oxygen radicals, 186, 660

Cisplatin Cisplatin -anti-idiotypic conjugate, in treatment of murine B-cell tumors pharmacological activity, 178, 374 preparation, 178, 373 Citrate buffer stock solutions, 182, 31 -phosphate buffer stock solutions, 182, 32 Citrate synthase assay, 188, 350 properties, 188, 352 purification from methylotrophs, 188, 351 ultramicroassay, 191, 548 Clastogenic factors assay, 186, 559 preparation, 186, 556 Cloning arachidonate 5-1ipoxygenase cDNA, 187, 491 avidin, 184, 71 DNA into pET vectors, 185, 78 genes in yeast by complementation, 194, 195 immunoglobulin genes, 178, 460 leukotriene A4 hydrolase cDNA, 187, 486 mammalian cells for recombinant protein production, 185, 568 molecular in analysis of nucleosome positioning, 170, 408 interstitial retinol-bindingprotein, 189, 207 retinoid receptors, 189, 224 nuclear retinoic acid receptors, 189, 256 prostaglandin endoperoxide synthases from mouse and sheep, 187, 469 protein gene from purified protein, 182, 738 rearranged Ig gene for production of chimeric antibody with antifibrin/tPA functions, 178, 517 renal cell lines, 191, 428 Spo- genes, 194, 123 sub-, see Subcloning Ty-induced mutations in yeast, 194, 357 in yeast artificial chromosome vectors, 194, 251

48 yeast genes by gap repair, 194, 815 yeast mtDNA in bacterial vectors, 194, 159 Clostridium histolyticum collagenase in dissociation of gastric mucosa into individual cells, 171, 448 Clostridium perfringens purification of endo-fl-galactosidase releasing Gal(al---~3)Gal, 179, 498 Cloud point detergents, 182, 244 Coagulant assay high-molecular-weight kininogen in plasma, 169, 278 Coagulation disseminated intravascular, role of oxygen radicals, 186, 656 CODATA Task Group on biological macromolecules, 183, 58 Cofactors nonprotein, analysis, 182. 626 reducing, oxidation by prostaglandin H synthase, 186, 286 Colicins bilayer spanning domains, prediction, 172, 439 hydrophobic moment, 172, 457 secretion from Escherichia coli, 185, 175 Collagen effect on citrated platelet-rich plasma, 169, 131 induced preformed platelet aggregates, adherence to endothelial cell monolayers, 169, 75 preparation for analysis of plateletsurface interactions, 169, 50 rat tail, preparation, 192, 386 solid-state 2H NMR, 176, 385 type IV differentiation marker for embryonal carcinoma cells, 189, 345 fibroblast attachment, enhancement by retinoic acid, 190, 87 Collagenase digestion of pancreas, 192, 190 expression control by retinoids, modulation of extracellular matrix, 190, 175 in hepatocyte preparation, 173, 566 in isolation of nonparenchymal rat liver cells, 190, 53

49 rat liver cells, 190, 50, 53 perfusion of kidney, 191, 227 liver, 190, 62 Collision-induced dissociation high-energy overview, 193, 240 spectra computer-aided interpretation, 193, 472 Leu/Ile and Lys/Gln differentiation, 193,468 and tandem MS, in peptide sequencing, 193,455 low-energy methods, 193, 251 multiply charged molecular ions, in analysis of oligopeptides and proteins, 193, 428 Colon cecum, electrical measurements, 192, 459 cell membranes, electrophysiology, 192, 482 electrical measurements, 192, 459 rat, short-circuit current determination in vivo. 192, 20 turtle apical ion channels, 192, 737 basolateral transporters, 192, 739 salt absorption, model, 192, 744 transepithelial transport, 192, 735 Colony screening for preparation of clone libraries in YAC vectors, 194, 265 yeast cell and nuclear fusion mutants, 194, 774 yeast secreting heterologous proteins, 194, 497 Colorimetric assay biotin-labeled probes after hybridization, 184, 596 dichloromethane dehalogenase, 188, 355 DNA-sequencing bands, 184, 612 enzymatic avidin and biotin, 184, 217 biotinylated DNA, 184, 611 3-hexulose-6-phosphate synthase, 188, 392 lipid content of glucose transporter preparations, 174, 43

Computer applications platelet lysosomal acid glycosidase secretion, 169, 338 proteins, 182, 57 Column switching in HPLC analysis of retinoids, 189, 146 Comamonas testosteroni

purification of quinohemoprotein alcohol dehydrogenase, 188, 37 Commelina communis

guard cell protoplasts, preparation, 174. 319 Complement C3 acceptors, identification, 184, 624 hemolytically active, biotinylation, 184, 621 C3b, avidin-linked oligomers with enhanced affinity for receptors, 184, 626 effect on endotoxin-induced circulatory shock, 186, 662 Complementation in cloning of genes in yeast, 194, 195 in genetic analysis of yeast, 194, 18 in identification of trans-acting components in histone RNA 3' processing, 181, 81 in test for fusion ability in yeast cell and nuclear mutants, 194, 780 Computer applications B-cell determinant prediction, 178, 586 distance geometry, 1"/7, 165 hydrophilicity analysis of protein antigenic segments, 178, 578 membrane fluorescence anisotropy data analysis, 172, 465 molecular dynamics simulation, 177, 204 molecular phylogenetic trees, 183, 599 NMR-related programs availability, summaries, and critiques, 177, 455 overview, 176, 477 for NMR resonance assignments, 177, 150 nucleic acid sequence alignment, 183, 331 divergence estimation, 183, 503 pattern detection and analysis, 183. 161 in patch-clamp recording of single-

Computer applications channel currents from human red cells, 173, 115 permeability coefficient calculation, 172, 274 peptide high-energy CID spectra interpretation, 193, 472 protein sequence alignment, 183, 331 sequence divergence estimation, 183, 503 solution structure determination from NMR data, 177, 218 structural analysis, 182, 751 RNA folding programs, 180, 267 RNA structure prediction pseudoknots, 180, 300; 183, 306 related considerations, 180, 262 secondary structure, 183, 279 from sequence, 180, 238 tertiary structure, 183, 318 sequence database searching, 183, 61 simultaneous sequence alignment and phylogeny reconstruction, 183, 637 solute movement modeling, 172, 280 steady-state rate equation generation for enzyme and carrier transport, 171, 164 T-cell epitope identification, 178, 617, 628 transport kinetics analysis, 172, 262 vesicle size distribution from photoelectron spectroscopy, 172, 330 volume change modeling, 172, 280 Conductance in analysis of ionic and nonelectrolyte permeation, 171, 705 in giant algal cells, analysis, 174, 417 K + in intestinal secretory cell membranes characterization, 192, 309 pharmacological properties, 192, 316 in Nitella membranes, determination, 174, 482 paracellular shunt in epithelia, determination, 191, 4 planar membranes, 171, 361 Conductometry in analysis of proton extrusion in yeast, 174, 601

50 Congenital abnormalities in platelet aggregation, 169, 133 Conglutinin purification from bovine serum, 179, 319 Conjugation yeast with bacteria, in transmission of plasmid DNA, 194, 187 Connectivities degenerate protons in heteronuclear NMR, 176, 110 dipolar, s e e Dipolar connectivities tracing along aliphatic side chains, 176, 162 Continuous flow tube methods in assay of anion, nonelectrolyte, and water transport, 173, 161 Coomassie Blue in colorimetric assay of proteins, 182, 62 properties, 182, 479 •staining of proteins in electrophoretic gels, 182, 478 Coomassie Blue R-250 in protein quantitation, 182, 487 Coomassie Brilliant Blue R-250 staining of proteins in SDS-PAGE gels, 182, 437 Copper alkaline, in colorimetric assay of proteins, 182, 57 -ascorbate system, radical damage, assay, 186, 167 chelatable by phenanthroline, assay, 186, 40 -histidine complexes, preparation, 168, 711 induction of yeast metallothionein promoter, levels for, selection, 185, 324 intracellular quenching of quin2, 172, 251 metabolism in humans, overview, 186, 38 -polyaminocarboxylate complexes, reactions with free radicals, 186, 119 staining of proteins in SDS-PAGE gels, 182, 437 Coprecipitation DNA and calcium phosphate, in DNA introduction into mammalian cells, 185, 528

51 Cornea bovine, endothelial cell culture and transport studies, 192, 571 Correlation times in NOE studies of ligand conformation and ligand-enzyme interactions. 177, 337 in quadrupolar ion NMR relaxation estimation, 171, 298 interpretation, 171, 306 Corticotropin-releasing factor cytotoxic analog, in study of target cells in mixed populations, 168, 41 fluorescent analog cell labeling, 168, 31 synthesis, 168, 31 Corticotropin-releasing factor receptors immunocytochemical images, 178, 219 immunocytochemical studies with anti-Id antibodies, 178, 216 Cortisol salivary, competitive-type immunoassays, 184, 492 Cost matrices in protein sequence comparison, 183, 135 COSY, s e e Homonuclear correlated spectroscopy Cotransfection CaPO4-mediated, in cotransformation of mammalian cells, 185, 563 Sf9 cells, 187,498 Coulometry titration of chloride, 173, 62 Counterstaining in in situ hybridization histochemistry with RNA probes, 168, 776 Coupling constants for phosphorylated amino acids in 3tp NMR study, 177, 270 Coupling rules for active transport and other coupled vectorial processes, utilization, 171, 145 Crambin adjacent peptide planes, relative orientations, 176,268 peptide backbone structure, solid-state NMR study. 176, 244

Crosslinking Crassulacean acid metabolism, associated coupling of water storage with active process, 174, 2O7 Creatine kinase bound reactants and products, equilibrium constant determination, 177, 364 interconversion rates, NMR analysis. 176, 297 isoenzymes, double-antibody immunoprecipitation assay, 182, 712 nuclear labels, indirect detection with spin echoes, 176, 351 3~p NMR, simulated and experimental spectra, comparison, 176, 299 CRF, s e e Corticotropin-releasing factor Critical micelle concentration detergents, 172, 528; 182, 244 Critical micelle temperature detergents, 182, 244 CRNA program prediction of optimal and suboptimal RNA secondary structure, 183, 282 Crosslinking anion-exchange channel in intact erythrocytes, 172, 625 band 3 protein oligomers, 173, 512 in characterization of bombesin-gastrin-releasingpeptide, 168,486 protein oligomeric structure, 191, 847 chromatin, psoralen method, 170, 155 covalent photoreactive biotinylated ADP analog to myosin ATPase site, 184, 412 in receptor identification, 191,634 DNA-protein identification in nucleohistone. 186. 540 in nuclei, 170. 390 on nucleosomes and soluble chromatin. 170, 389 to partly depurinated DNA, 170, 388 by ultraviolet light, 170, 391 histones. 170, 549 hormone-receptor coupling, 172. 605 internal erythrocyte plasmalemmal proteins, 184, 158

Crosslinking intrasubunit, in analysis of membrane proteins, 172, 614 macromolecules on intact erythrocyte membranes, 184, 158 for membrane protein analysis, 172, 584 photoaffinity, in analysis of RNAprotein interactions, 180, 383 principles, 172, 611 proteins as evidence of multienzyme complexes, 182,547 by N~-(y-glutamic)lysine, 182, 636 in quaternary structure studies, 172, 612 rabbit muscle aldolase, 172, 623 sarcoplasmic reticulum membrane Ca2+ATPase, 172, 606 sterically enforced reactions, 172, 605 topographical studies, 172, 608 ultraviolet-induced proteins to precursor RNA, 181, 68 RNA to proteins in vivo, 180, 410 RNA to RNA, analysis, 180, 418 vectorial, and biotinylation, purified proteins, 184, 159 Crosslinking reagents membrane impermeant, for analysis of membrane proteins, 172, 609 for protein-nucleic acid reactions, 172, 604 for proteins and protein complexes, 172, 586 Cryofracturing frozen hydrated bulk plant specimens for X-ray microanalysis, 174, 270 Cryoultramicrotomy specimen preparation for electron probe X-ray microanalysis, 172, 208 Crystallization cellular retinol-bindingprotein, 189, 519 nucleosome core particles, 170, 420 protein-DNA complexes, 182, 658 proteins, 182, 646 retinoids, 189, 18 retinol-bindingprotein, 189, 283 transaldolase from Candida utilis, 182, 788 Culture media for Bacillus subtilis inducibly expressing regulatory genes, 185, 227

52 secreting heterologous proteins, 185, 222 chlorate-containing, cells incubated in, undersulfation of proteoglycans, 179, 429 for Escherichia coli strains BL21 and HMS174, 185, 77 for islet cells, optimal glucose levels, 192, 216 for mammalian cells producing recombinant proteins, selection, 185, 575 for Neurospora in analysis of K+-H + transport, 174, 658 for Saccharomyces cerevisiae sporulation, 194, 94 for Schizosaccharomyces pombe, 194, 800 serum-free, for mammalian cells producing recombinant proteins, 185, 576 for Spodoptera frugiperda Sf9 cells, 182, 121 for submerged keratinocyte culture, 190, 21 sulfate-depleted, cells incubated in, undersulfation of proteoglycans, 179, 428 for three-dimensional pituitary cell cultures, 168, 49 for yeast growth, 182, 155; 194, 12 for yeast sporulation, 194, 17, 94 Cumyl hydroperoxide demethylation of N-methylaniline, assay, 186, 274 hydroxylation of toluene, assay, 186, 275 Cuvettes glass, see Glass cuvettes Cyanamide in preparation of leukotriene B4 immunogens, 187, 123 leukotriene C4 immunogens, 187, 118 Cyanoborohydride in assay of carbonyl content in oxidatively modified proteins, 186, 476 Cyanogen bromide activation of Sepharose, 184, 185 cleavage of proteins for N-terminal sequence analysis, 182, 610 coupling of band 3 protein, 173, 507 digestion of proteins for MS analysis, 193, 371

53 Cyclic AMP accumulation in gastric parietal cells, assay, 191,649 3H-labeled assay in blood from transfused rabbit, 169, 468 labeled platelets, 169,460 in detection and assay of circulating prostaglandin 12, 169, 462 in platelet studies in vivo, 169, 466 messenger system and Ca2÷ messenger system integration in cellular response, 191, 667 interrelationships, 191, 666 model of epithelial cell stimulussecretion coupling, 191, 662 metabolism, correlation with gastric parietal cell function, 191,659 in nephron segments, radioimmunoassay, 191, 310 platelet, assay by prelabeling techniques, 169, 455 trimethylsilyl derivatives, preparation, 193,795 Cyclic GMP in nephron segments, radioimmunoassay, 191, 310 Cyclic nucleotides in nephron segments, radioimmunoassay, 191, 310 related measurements, in gastric parietal cells, 191, 649 Cyclohexanone 1,2-monooxygenase assay, 188, 71 properties, 188, 74 purification from A c i n e b a c t e r NCIMB 9871, 188, 72 l-Cyclohexyl-3-(2-morpholinoethyl)carbodiimide metho-p-toluene sulfonate in structural probing of native snRNA, 180, 215,221 Cyclooxygenase in assay of apoprostaglandin endoperoxide synthase, 187, 479 initiation assay for hydroperoxides, 186, 431 Cyclopentanone 1,2-monooxygenase assay, 188, 77

Cytochrome c properties, 188, 80 purification from P s e u d o m o n a s NCIMB 9872, 188, 78 8,5'-Cyclopurine-2'-deoxynucleosides in DNA, analysis, 186, 538 Cysteine alkylation for amino acid analysis, 182, 598 cellular production of sulfate from, 179, 434 oxidation by performic acid, 182, 598 in proteins, spin labeling, 177, 116 reduction for amino acid analysis, 182, 598 spin system identification, 177, 138 thiyl radicals, generation by peroxidases, 186, 319 Cystic fibrosis phenotype, airway epithelial cells with persistence in, transfolmation, 192, 565 Cystine exodus from lysosomes, analysis, 174, 154 oxidation by performic acid, 182, 598 thiol promoted uptake, assay, 186, 707 Cystinosis related cystine exodus from lysosomes, analysis, 174, 154 Cytidine Y-monophosphate, see Y-Cytidylic acid Cytidine 5'-monophosphate trimethylsilyl derivatives, preparation for MS analysis, 193,795 Cytidine 5'-monophospho-N-acetylneuraminic acid preparation, 179, 275 3'-Cytidylic acid -ribonuclease A complex, 3tp NMR studies, 177, 313 Cytochalasin B binding by glucose transporter, 174, 44 Cytochemistry affinity, see Affinity cytochemistry Cytochrome b bilayer spanning domains, prediction, 172, 439 Cytochrome c reduction in assay of

Cytochrome c HL60 cell differentiation, 190, 126 phthalate dioxygenase reductase, 188, 62 reductase component of naphthalene dioxygenase, 188, 48 toluene dioxygenase, 188, 41 dye-mediated, 186, 638 soluble assay, 188, 290 identification in methanotrophs, 188, 296 properties, 188, 295 purification from Methylomonas A4, 188, 291 superfamily alignment, 183, 468 from various species, phylogenetic relationships, 183, 685 Cytochrome cH Hyphomicrobium X assay, 188, 304 properties, 188, 307 purification, 188, 305 Methylobacterium extorquens AM1 assay, 188, 299 properties, 188, 303 purification, 188, 301 Cytochrome CL Hyphomicrobium X assay, 188, 304 properties, 188, 307 purification, 188, 305 Methylobacterium extorquens AM1 assay, 188, 299 properties, 188, 302 purification, 188, 301 Cytochrome-c oxidase mitochondrial marker, assay, 182, 214 reconstituted, H ÷ and charge stoichiometry, 172, 154 Cytochrome f hydropathy plot, effect of averaging window, 172, 448 Cytochrome oxidase, see Cytochrome-c oxidase Cytochrome P450 form 5, N-terminal sequence analysis, 187, 265 ocular assay, 187, 375 metabolism of arachidonate, 187, 372

54 peroxygenase activity assay, 186, 273 role of alkoxy and oxygen radicals, 186, 277 reductive cleavage of hydroperoxides, 186, 278 Cytochrome P450LTa characteristics, 187, 319 Cytochrome P450 arachidonate oxygenase microsomal assay, 187, 386 biochemical characterization, 187, 385 reconstitution, 187, 392 Cytochrome P450 prostaglandin to-hydroxylase rabbit lung, pregnancy-inducible assay, 187, 260 N-terminal sequence analysis, 187, 265 purification and properties, 187, 253 purified, detergent removal, 187, 258 Cytoplasm Ca2÷ assay in living cells, 173, 745 dye loading, 173, 763 giant algal cell, perfusion, 174, 437 pH in Chara cells, effect on ion transport, 174, 473 in lymphocytes, measurement, in detection of Na÷/H + exchange, 173, 777 and proton pumping, assay in cell culture, 174, 146 preparation with ribonucleoside-vanadyl complexes, 180, 8 RNA isolation, 180, 6 Cytoplasmic polyhedrosis virus transcription, in synthesis of capped mRNA in vitro, 180, 167 Cytosine in DNA, modification, 184, 600 in RNA, modification by hydrazine, 180, 374 Cytosine 1-fl-D-arabinofuranoside -anti-idiotypic conjugate, in treatment of murine B-cell tumors pharmacological activity, 178, 374 preparation, 178, 372 Cytoskeleton erythrocyte, shell preparation and coating with lipids, 173, 394

55

11-Dehydrothromboxane B2

phospholipid membrane-coated characterization, 173, 395 preparation by erythrocyte reconstitution, 173, 392 protein thiols, quinone-induced depletion, role of glutathione, 186, 635 yeast, structural elements, detection, 194, 600 Cytosol atrial and ventricular myocytes, assay of Ca 2~, 173, 674 components, resealable membranes depleted of, preparation, 173, 346 CVI cell, preparation, 170, 47 embryonal carcinoma cell, preparation, 190, 150 free Ca 2+ levels assay with quin2, 172, 230 hormone effects, 173, 541 hepatic, purification of epoxide and leukotriene A4 hydrolases, 187, 328 marker enzyme lactate dehydrogenase, assay, 182, 220 marker RNA, assay, 182, 221 pH measurement, 172, 85 porcine leukocyte, fraction preparation. 187, 340 Cytotoxicity anti-Id antibody-neocarzinostatin conjugates to anti-DNA Id-positive cells, 178,428 monoclonal antibody-mutant diphtheria toxin immunotoxins, assay, 178,415

D Dark adaptation in analysis of retinoid binding by interstitial retinol-binding protein, 189, 326 Data analysis curve-fitting for platelet Ca pool 45Ca content, 169, 373 for in situ hybridization, graphical and statistical methods, 168, 822 Databases, s e e also s p e c i f i c d a t a b a s e s biological macromolecules and components, 183, 54 eukaryotic promoters, 183, 26

evolutionary trends, 183, 51 NMR, 176, 62 nucleic acid sequences protein similarity detection with, 183. 111 searches, 183, 99 sources, 182, 775 profile analysis, 183, 146 protein sequences searches, 183, 99 sources, 182, 775 rate constants of oxygen radicals in solution, 186, 113 restriction enzymes, 183, 26 retrieval capacity, 183, 50 Daunomycin, s e e Daunorubicin Daunorubicin -anti-idiotypic conjugate, in treatment of murine B-cell tumors pharmacological activity, 178. 374 preparation, 178, 371 Deadenylylation glutamine synthetase, 182, 799 DEAE-cellulose, s e e Diethylaminoethylcellulose DEAE-dextran, s e e Detaxtran DEAE-Sephadex, s e e Polidexide Deamido-thyrotropin-releasing hormone radioimmunoassay, 168. 572 Decoupling composite pulse schemes for, mixing based on, 176, 156 Degassing freeze-thaw, in NMR sample preparation, 176, 88 Deglycosylation glycoproteins, 182, 537 Dehydration artery segments, 169, 62 in orientation of native and synthetic membranes, 172,401 -rehydration, multilamellar vesicles. 171, 199 Dehydrogenases mammalian intramitochondrial effects of Ca ~, 174, 95 properties, 174, 95 1l-Dehydrothromboxane B2 chemical properties, 187, 35 GC/MS assay, 187, 42

11-Dehydrothromboxane B2 in plasma and urine, assay, 187, 39 radioimmunoassay, 187, 34 Dejellying X e n o p u s eggs, 170, 615 Delayed luminescence anisotropy in measurement of rotational diffusion in membranes, 172, 476 Demethylation N-methylaniline by cumyl hydroperoxide, assay, 186, 274 Denaturation heat- and pH-dependent, in enzyme purification, 182, 298 Denaturing agents protein chromatofocusing in, 182, 390 Density based separation of islets from exocrine tissue, 192, 195 cell, aspects relevant to gradient centrifugation, 171, 464 cell suspension, effect on transport parameters in yeast, 174, 584 electric current, extracellular, in epithelia, electrophysiological scanning, 171, 793 erythrocyte ghosts, in barrier assay of hemolytic hole size, 173, 365 gradients in cell separation after elutriation, 171, 490 for velocity sedimentation, 171, 468 Nycodenz solutions, determination from refractive index, 171, 510 Deoxycholate and trichloroacetic acid, precipitation of proteins, 182, 593 2'-Deoxycytidine photoproducts, isolation and characterization, 186, 508 sensitized photooxidation, mechanism, 186, 510 Deoxyribonuclease I digestion of nuclei, 170, 326 nucleosomes, 170, 264 footprinting, in analysis of reconstituted nucleosome core particle authenticity, 170, 580 hypersensitive sites in chromatin, analysis, 170, 269

56 sensitivity of nucleosome core particles, internal structure, 170, 107 Deoxyribonuclease II digestion of nucleosomes, 170, 264 Deoxyribonucleoproteins isolation from wheat germ, 170, 440 wheat leaf mesophyll cells, 170, 441 yeast, 170, 439 Deoxyribose in assay of free hydroxyl radicals and site-specific .OH production, 186, 26 Dephosphorylation inositol phosphate, 168, 343 -phosphorylation ratio, peptides, estimation, 193, 489 Deplasmolysis definition, 174, 233 Depolymerization cytoskeletal material, in isolation of brush border membranes, 192, 395 Deposition platelet, definition, 169, 40 thrombi on canine ex oivo polymeric shunt surfaces, 169, 103 transient, platelets, 169, 99 Derivatization chemical, for mass spectrometry, 193, 314 with vapor-phase reagents, 193, 323 Desalting photoproteins, 172, 170 purified nucleoplasmin, 170, 622 Desferal radioprotective effects in mouse, 186, 686 in spin trapping of free radicals, 186, 131 Desferrioxamine and MnO2, derived superoxide dismutase mimic preparation, 186, 243 properties, 186, 244 Desmosine assay in skin, 190, 354 Desorption field glycosphingolipids, 193, 722 mass reference standards, 193, 310 laser, see Laser desorption-ionization

57 Desquamation retinoid effects, 190, 81 Detaxtran mediated transfection of COS-I cells, 185, 507 mammalian cells, 185, 530 Detergents, s e e also s p e c i f i c d e t e r g e n t s additives for isoelectric focusing of proteins, 182, 466 aggregation principles, 182, 240 critical micelle concentration, 172, 528 in isolation of sea urchin embryo nuclei, 170, 437 -membrane protein complexes, size and shape, hydrodynamic studies, 172, 696 nonionic, prevention of rRNA terminal maturase loss in solution, 181, 370 permeabilization of mammalian cells to macromolecules, 171, 866 in preparation of fusogenic reconstituted Sendal virus envelope or hybrid vesicles, 171, 842 nuclei, 180, 11 properties, 182, 244 protein-bound, assay, 172, 526 for protein purification overview, 182, 239 selection, 182, 251 removal from membrane proteins, 182, 277 purified cytochrome P450 prostaglandin o~-hydroxylase, 187, 258 in removal of plasma membrane, 171, 820 solubilization of band 3 protein, 173, 499 lipids in preparation of unilamellar vesicles, 171, 205 membrane proteins, 182, 253 pulmonary microsomes, 187, 254 solutions, associated critical phenomena, 172, 530 structure, 182, 239 Deuteration proteins for ~H NMR, 177, 23 retinoids, 189, 95 in solvent suppression for NMR, 176, 66

Dialysis staphylococcus nuclease for residue-type assignment of NOE cross-peaks, 177, 284 Deuterium content, calculation, 193, 336 exchange methods for electron and chemical ionization MS, 193, 332 for FAB/MS and liquid secondary ion MS, 193, 334 Dextran coated charcoal, in cellular retinoidbinding assays, 190, 152 fluorescein isothiocyanate-labeled in assay of proton pumping in lysosomes and related organelles, 174, 131 as pH indicator, 174, 137 in tracer studies of endocytosis. 191, 559 fluorescent, exocytosis, 172, 56 -polyethylene glycol-water system for partition of ceils and organelles, 171, 532 Dextran sulfate permeabilization of mammalian cells to macromolecules, 171,867 Diacylglycerides benzoylation, 187, 207 purification, 187, 207 Diacylglycerol and Caz* ionophore, synergistic effects on platelet secretion, 169, 436 conversion to arachidonate, stearate, and glycerol, inhibition, 191, 690 disposition on stimulation of phosphatidylinositol breakdown, 191, 688 Diacylglycerol lipase, see Lipoprotein lipase Dialysis in detergent removal from membrane proteins, 182, 279 flow, in determination of lipophilic ion distribution, 172, 67 micro-, see Microdialysis in preparative-scale transport-specific fractionation, 172, 47 in protein concentration and solute removal, 182, 72

Dialysis salt gradient, in reconstitution of nucleosome cores, 170, 576 4',6'-Diamidino-2-phenylindoledihydrochloride in staining of yeast nuclei, 194, 821 4,6-Diamino-5-N-formamidopyrimidine detection, 186, 517 Diazirines general properties, 172, 643 Diazobenzoylbiocytin biotinylation of proteins, 184, 151 interaction with DNA, 184, 610 Diazonium coupling reaction for sulfinic acid, 186, 144 in synthesis of [170] or [1sO]hydroxylenriched 4-hydroxyphenylacetate, 188, 108 1-Diazo-2-oxo-4-(S-2,4-dinitrophenyl)mercaptobutane affinity labels for catalytic antibodies, 178,558 l-Diazo-2-oxo-6-(S-2,4-dinitrophenyl)mercaptohexane affinity labels for catalytic antibodies, 178, 560 1-Diazo-2-oxo-5-(S-2,4-dinitrophenyl)mercaptopentane affinity labels for catalytic antibodies, 178, 559 Dichloromethane dehalogenase assay, 188, 355 properties, 188, 360 purification from H y p h o m i c r o b i u m DM2, 188, 357 N,N'-Dicyclohexylcarbodiimide effect on Rhodotorula plasma mem-

brane, 174, 647 3,4-Didehydroretinol, see Vitamin A2 3,4-Didehydroretinyl esters radioactive, analysis, 190, 213 synthesis by organ-cultured human skin, 190, 210 DIDIT membrane-impermeant crosslinking reagent, 172, 618 Dienes conjugation, in assay of lipid peroxides in human body fluids, 186, 66

58 Dienols and methyl linoleate hydroperoxides, HPLC, 186, 381 Diet for preparation of vitamin A-deficient rats and mice, 190, 229 Diethylaminoethyl-cellulose benzoylated, in ion-exchange chromatography of RNA, 180, 28 in ion-exchange chromatography of RNA, 180, 26 Diethyl oxocarbodicyanine in measurement of potential in cells and vesicles, 191, 477 Diethyl pyrocarbonate, see Pyrocarbonic acid diethyl ester Differentiation defined renal tubule cells in vitro, requirements, 191, 415 embryonal carcinoma cells assessment, 189, 341 induction, 189, 340 markers, 189, 344 mRNA analyses, 189, 346 epithelial analysis with porous-bottom culture dishes, 171, 736 in hamster trachea, role of retinoids, 190, 91 as measure of vitamin A status in human populations, 190, 247 role of retinoids, 190, 19 HL60 assays, 190, 122 induction by gangliosides, 179, 538 retinoic acid-induced, analysis, 190, 120 keratinocytes, vitamin A-mediated regulation, 190, 18 teratocarcinoma F9 cells, 190, 112 Diffusion bulk, D20 across membranes, 173, 199 electro-, see Electrodiffusion rotational, in membranes, fluorescence and phosphorescence measurements, 172, 472 simple, as model of membrane transport, 171, 33 translational

59 in membranes, fluorescence and phosphorescence measurements, 172, 495 proteins, pulse-gradient spin-echo NMR measurement, 176, 418 water across red cell membranes, 173, 192 Diffusion coefficient membrane protein-detergent complexes, 172, 698 Diffusion potential driven CI- uptake by intestinal membrane vesicles, 192, 407 electrical, in selective amplification of isotope uptake by membrane vesicles, 172, 155 Diffusivity tissue, absolute value and physiological meaning, 174, 193 Digalacturonic acid enzymatic digestion, 179, 567 isolation, 179, 568 proteinase inhibitor inducing activities, 179, 569 A4~-Digalacturonic acid enzymatic digestion, 179, 567 isolation, 179, 568 proteinase inhibitor inducing activities, 179, 569 Digital image processor in measurement of fluorescence, 192, 43 Digitonin permeabilization of cells for determination of subcellular isotope distribution, 173, 553 pancreatic acinar cells, 192,283,292 parietal cells, 192, 108 Diglycosylceramides perbenzoylation, 172, 544 Diheptanoylphosphatidylcholine interaction with bacteriorhodopsin and lipid bilayer, 171, 272 Dihydrofolate reductase expression plasmid, CaPO4 cotransfection of mammalian cells, 185, 563 gene amplification, methotrexate resistance selection for, 185, 544 Dihydrolipoamide dehydrogenase catalyzed reactions, role of thiyl radicals, 186, 328

Dimerization Dihydroorotase -carbamoyl-phosphate synthase-aspartate transcarbamoylase, in gene selection and coamplification. 185, 550 7,8-Dihydro-8-oxoadenine detection, 186, 517 Dihydroxyacetone synthase, s e e Formaldehyde transketolase Dihydroxy aromatic compounds 170- or 1sO-enriched, synthesis. 188, 107 purification from B r e v i b a c t e r i u m f u s c u r e , 188, 112 Dihydroxyboryl columns in affinity chromatography of RNA, 180, 38 cis- 1,2-Dihydroxycyclohexa-3,5-diene (NAD) oxidoreductase s e e c/s-Benzene dihydrodiol dehydrogenase 5,6-Dihydroxyeicosatetraenoic acid formation from leukotriene A4, catalyzing enzymes, purification and properties, 187, 324 5(S), 12(S)-Dihydroxy-6,10-trans-8,14-ciseicosatetraenoic acid preparation, 187, 305 12S-20-Dihydroxyeicosatetraenoic acid production by neutrophils, assay, 187, 594 Dihydroxyeicosatrienoic acids tsO2-1abeled, preparation, 187, 184 production by hepatic microsomes, assay, 187, 182 stable isotope-dilution MS assay, 187. 175 Diisopropyl fluorophosphate, s e e Isoflurophate Dilution effect on purification of multienzyme complexes, 182, 540 isotope, s e e Isotope dilution multiple indicator, s e e Multiple indicator dilution Dilution potential in analysis of transepithelial ion fluxes in teleost intestine, 192, 747 Dimerization antibody McPC603 variable domains, 182, 138

7,12-Dimethylbenz[a]anthracene 7,12-Dimethylbenz[a]anthracene, s e e 9,10Dimethyl-1,2-benzanthracene 9,10-Dimethyl-l,2-benzanthracene induction of breast cancer, 190, 400 Dimethyl bisdithiopropionimidate in crosslinking of histones, 170, 550 2,2-Dimethyloxirane hydrolysis by epoxide hydrolase, mechanistic studies, 177, 387 Dimethyl suberimidate in crosslinking of histones, 170, 550 Dimethyl sulfate in structural probing of native snRNA, 180, 214, 221 Dimethyl sulfoxide in detection and assay of hydroxyl radicals, 186, 137 Dimyristoylphosphatidylcholine and cholesterol, vesicles, spontaneous insertion of membrane proteins, 171, 261 and myristate, vesicles, spontaneous insertion of membrane proteins, 171, 258 purification, 171, 255 vesicles, preparation, 171, 256 2,4-Dinitrobenzoic acid 7-hydroxycoumarin ester substrate for catalytic antibody, synthesis, 178, 564 (N-2,4-Dinitrophenyl)-2-aminoethyl 2-(1,3dioxolanyl)ethyl disulfide affinity labels for catalytic antibodies, 178, 555 (N-2,4-Dinitrophenyl)-2-aminoethyl 3-(1,3dioxolanyl)propyl disulfide affinity labels for catalytic antibodies, 178, 557 (N-2,4-Dinitrophenyl)-2-aminoethyl 4oxobutyl disulfide affinity labels for catalytic antibodies, 178, 557 (N-2,4-Dinitrophenyl)-2-aminoethyl 3oxopropyl disulfide affinity labels for catalytic antibodies, 178, 556 (N-2,4-Dinitrophenyl)-3-aminopropanoic acid 7-hydroxycoumarin ester substrate for catalytic antibody, synthesis, 178,564

60 Dinitrophenylation monoclonal antibodies, 184, 348 N-Dinitrophenyl-S-carboxymethylaminothiols HPLC, 186, 707 2,4-Dinitrophenylhydrazine in assay of carbonyl content of oxidatively modified proteins, 186, 471 Dinitrophenylhydrazone formation, in assay of aldehydes, 186, 415 4-(S-2,4-Dinitrophenyl)mercaptobutanoic acid affinity labels for catalytic antibodies, 178, 559 5-(S-2,4-Dinitrophenyl)mercaptopentanoic acid affinity labels for catalytic antibodies, 178, 559 3-(S-2,4-Dinitrophenyl)mercaptopropanoic acid affinity labels for catalytic antibodies, 178, 557 2,3-Dinor-thromboxane B2 GC/MS assay, 187, 42 Diode array detectors for HPLC of proteins, 182, 398 Dipeptidyl aminopeptidase B yeast vacuole, assay, 194, 656 Diphenylpicrylhydrazyl oxidation of tocopherols, 186, 199 Diphenyl-1-pyrenylphosphine in assay of hydroperoxides, 186, 157 preparation, 186, 158 Diphtheria toxin based idiotype-specific immunotoxins for B-cell neoplasms, preparation, 178, 358 characteristics, 178, 407 mutant derived immunotoxins ADP-ribosylation assay, 178, 414 cytotoxicity assay, 178, 415 inactivation of protein synthesis, kinetics, 178, 418 synthesis and purification, 178, 411 preparation, 178, 407 Dipolar connectivities intramolecular, determination with

61 selective relaxation method, 176, 197 Dipolar interactions intramolecular, determination with selective relaxation method, 176, 198 3,3'-Dipropyloxadicarbocyanine in assay of membrane potential of mufine ascites tumor cells, 172, 95 3,3'-Dipropylthiacarbocyanine in determination of yeast membrane potential, 174, 615 3,3 '-Dipropylthiadicarbocyanine in determination of membrane potential in murine ascites tumor cells, 172, 95 yeast, 174, 615 3,3'-Dipropylthiadicarbocyanine iodide in measurement of potential in cells and vesicles, 191, 477 Disaccharides ions, preparation for tandem MS analysis. 193, 615 Discrete Fourier transform as NMR spectral estimate, 176, 217 Disodium 5'-GMP self-assembled structures, 3tp NMR inversion transfer study, 176, 322 Dispersion media for cell separation by elutriation, 171, 485 Dissection immuno-, s e e Immunodissection in isolation of Ca2+-tolerant atrial and ventricular myocytes, 173,665 Malpighian tubules, 192,620 micro-, s e e Microdissection yeast asci, 194, 21 Dissipation function for analysis of biological transport, 171, 400 Dissociability pharmacologic specificity of ligand and elicited anti-ld antibody, 178, 165 Dissociation collision-induced, s e e Collision-induced dissociation Distance geometry atom representations, 177, 165 bounds, 177, 174

Diuretics computational requirements and performance, 177, 188 conformational searches, general features, 177, 160 constraints, 177, 161 data input file generation, 177. 169 generated structures accuracy, 177, 196 analysis, 177, 192 and molecular dynamics simulation, in structural analysis, 177.211 optimization, 177, 180 parameter input file, 177, 169 refinement, 177, 190 structural data reporting, 177, 200 structure generation from NMR studies general features, 177, 162 initial generation, 177. 178 parameters, 177, 187 Distillation glycosphingolipid mixtures, 193, 632 Disulfide bonds peptide, reduction, 193, 328 in proteins assignment, 168, 96 localization, 193, 374 in recombinant human erythropoietin. analysis, 193, 535 Disulfide radicals anions, chemical properties, 186. 175 Disulfides glutathione-protein mixed, quinoneinduced formation, 186, 630 and thiols, interconversion, 193. 375 Dithiobis(succinimidyl propionate) in crosslinking of histones, 170, 550 Dithioerythritol -dithiothreitol mixture, mass spectra. 193, 878 Dithiothreitol -dithioerythritol mixture, mass spectra. 193, 878 Diuretics furosemide-related chemistry, 191,781 evaluation in v i t r o , 191, 790 mechanism, 191,784 metabolism, 191, 789 structure-activity relationships. 191, 783

DNA DNA -amino acid crosslinks, GC/MS, 193, 850 bases free, GC/MS, 193, 846 modified, identification, 193, 853 binding assay, 182, 527 binding proteins affinity chromatography, 182, 521 OBF1, purification, 182, 524 reverse biochemical analysis, 194, 529 biotinylated addition to solid matrix, 184, 337 enzyme-mediated colorimetric detection, 184, 611 biotinylation, 184, 331 tert-butyldimethylsilyl derivatives, preparation, 193, 861 cellular, flow cytometric assay, 171, 561 in chromatin cleavage, 170, 410 lysine residues binding, radiolabeling, 170, 369 chromatin assembly on, 170, 608 chromosome-sized, preparation in agarose, 194, 60 circular, single-site labeling, 170, 604 cloning into pET vectors, 185, 78 complementary arachidonate 5-1ipoxygenase, clone isolation, 187, 492 avocado sunblotch viroid, clone preparation, 181, 584 end-labeled, in nuclease protection assays, 168, 639 genes expression vectors, 185, 495 glucocorticoid induction, 185, 499 glucose-H + cotransporter-encoding clone, identification, 174, 401 to lg mRNA, gel electrophoresis, 178, 98 leukotriene A4 hydrolase, cloning, 187, 486 lucerne transient streak virus virusoid, clone preparation, 181, 584 plasmid-borne acquisition by mail, 194, 214 applications, 194, 224

62 construction de novo, 194, 216 incompleteness, 194, 229 probes for detection of neuropeptide mRNA, 168, 702 for RNA intron mapping, preparation, 180, 327 cotransfected, amplification, 185, 541 crosslinked, spreading, 170, 158 8,5'-cyclopurine-2'-deoxynucleosidesin, analysis, 186, 538 cytosine residue modification, 184, 600 denatured regions, enhancement for electron microscopy, 170, 160 denatured, uncrosslinked, depletion from crosslinked samples, 170, 394 derivatization, 186, 533 detection with biotinylated nucleotides, 184, 561 digestion by nuclease SI, 170, 287 direct incorporation of biotin, 184, 608 direct transfer into mammalian cells, 185, 495 -DNA crosslinks formation, psoralen method, 170, 156 purification, 170, 157 double-strand breaks, delivery by pGAL-HO, 194, 140 double-stranded electron microscopy, 180, 252 thermal stability, 180, 240 enhancers assembly for control of transferred gene expression, 185, 512 cellular, assembly, 185, 520 viral, assembly, 185, 514 wild-type, analysis with enhancer/ promoter chimeras, 185, 514 -Escherichia coli Err7° complex, conformational changes, 185, 40 exogenous, introduction into yeast cells, 194, 182 expression cassettes GAL4, construction, function verification, and usage, 185, 297 glyceraldehyde-3-phosphate dehydrogenase-derived for constitutive synthesis of heterologous proteins, 185, 341 plasmid construction, 185, 344

63 for heterologous gene expression in yeast, 185, 231 expression in Escherichia coli by infection with bacteriophage CE6,185, 83 IPTG induction of BL21(DE3) or HMS174(DE3), 185, 79 foreign, expression in hgtl 1 recombinants, 194, 230 fragments blotting to DBM paper, 170, 334 gel analysis, 170, 332 histone transfer to, in nueleosome and chromatin assembly in vitro, 170, 578 hybridization, 170, 335 relative affinity of histones for, measurement, 170, 642 separation, 170, 289 G A P D H terminator, fragment preparation, 185, 343 genetically active, synthesis, 182,786 genomic divergence through gene rearrangement, 183, 428 expression unit integration, 185,425 human, high-molecular-weight isolation, 194, 255 partial digestion, 194, 257 hydrolysates, derivatization for GC/MS, 193, 845 hydrolysis enzymatic, for mass spectrometry, 193, 782 for GC/MS, 193, 843 for isotope dilution GC/MS, 193, 860 hydroperoxides in, functional determination, 186, 519 8-hydroxy-2'-deoxyguanosine in, HPLC with electrochemical detection, 186, 521 introduction into mammalian cells, 185, 527 labeling with biotinylated nucleotides, 184, 561 at 5' end, 1711, 633 lac operator, in expression of heterologous genes in Bacillus subtilis, 185, 203

DNA length, effect on nucleosome reconstitution, 170,636 libraries computer searches, 183, 91 genomic, plating with amplification. 18:l, 746 MA Ta2 operator clones with, screening, 185, 361 insertion into promoters procedure, 185, 360 site identification, 185, 357 preparation, 185, 358 methylated bases, quantitative analysis, 193, 778 5-methylcytosine in, isotope dilution GC/MS, 193, 857 mitochondrial sequence branching, in analysis of human evolution, 183, 563 yeast, intron description, 181, 540 mononucleosomal, cloning and sequencing to determine nucleosome positions, 170, 414 nascent, 3' and 5' ends, distance to first nucleosome on arms of replication fork, 170, 308 nuclear flow cytometric assay, 171,561 and homogenates, assay, 170, 322 movement during mitosis in yeast. digital imaging microscopy, 194, 750 nuclease specificity, 170, 292 nucleosides, GC/MS, 193, 848 nucleosome alignment, 170, 594 nucleosome core particle assembly on, 170, 493 ovine vesicular gland hgtl0 libraries preparation, 187, 471 screening, 187, 476 oxidative damage photodynamic studies, 186, 502 selected-ion MS assays, 186, 530 in vivo, 186, 25 plasmid, see Plasmids prenucleosomal identification, 170, 307 nucleosome structure and spacing in absence of, 170, 310

DNA preparation for mass spectrometry, 193, 782 for 5-methylcytosine determination, 193, 858 primers for primer extension analysis of RNA, selection and preparation oligonucleotides, 180, 352 restriction fragments, 180, 354 strand separation, 180, 359 promoters ADH2 effects of ADRI overproduction, 185, 315 in expression of heterologous proteins in Saccharomyces cerevisiae, 185, 308 plasmids with, construction, 185, 3O9 yeast expression plasmids with, construction, 185, 311 bacteriophage T7, linked pre-tRNA gene, construction, 180, 64 constitutive, for expression of cDNA genes in mammalian cells, 185, 496 for control of expression of transferred genes, assembly, 185, 512 -enhancer chimeras, in analysis of wild-type enhancers and promoters, 185, 514 Escherichia coli trp in direct expression of proteins, 185, 54 induction, 185, 57 eukaryotic, database, 183, 26 GAL in assay of yeast mRNA decay rates, 194, 421 high-copy, in yeast, regulated GAL4 expression cassettes for, 185, 297 in regulatable expression of cloned genes in yeast, 194, 379 GAPDH length and orientation, effects on expression of heterologous proteins, 185, 345 preparation, 185, 343 heat shock, for expression of cDNA

64 genes in mammalian cells, 185, 498 for heterologous gene expression in yeast, 185, 425 -heterologous gene junctions in yeast, 185, 427 inducible, for expression of cDNA genes in mammalian cells, 185, 497 interferon-B, for expression of cDNA genes in mammalian cells, 185, 497 lac, in vectors for transcription optimization in Escherichia coli, 182, 96 metallothionein, for expression of cDNA genes in mammalian cells, 185,498 PGK, high-efficiency yeast expression vector based on, 185, 329 recognition by Escherichia coli Eo"7° regulatable, in yeast vectors for expression of cloned genes, 194, 373 spac-1, induction ratio, 185, 226 strong genetic fusion to, 182, 93 inducible, control of staphylococcal nuclease gene, 177, 76 tac, in vectors for transcription optimization in Escherichia coli, 182, 96 temperature-regulated applications, 185, 365 construction, 185, 357 testing, 185, 362 wild-type, analysis with enhancer/ promoter chimeras, 185, 514 yeast copper metallothionein, for heterologous gene expression, 185, 319 for protein production by secretion, 194, 492 superimposition of temperature regulation, 185, 351 -protein complexes analysis, 170, 395 crystallization, 182, 658 electrophoresis, 170, 116 footprinting with y-rays, 186, 545

65 recovery, 170, 392 structure, 3ip NMR analysis, 177, 310 -protein crosslinking in nuclei, 170, 390 on nucleosomes and soluble chromatin, 170, 389 to partly depurinated DNA, 170, 388 polypeptide domains, mapping and identification, 170, 403 sites labeling with [32P]ATP, 170, 406 mapping on DNA fragment, 170, 401 by ultraviolet light, 170, 391 -protein crosslinks identification in nucleohistone, 186, 540 in nucleoprotein, GC/MS-SIM, 193, 855 purification from nuclear lysate, 170, 286 and redigestion by restriction enzymes, 170, 330 purified, restriction endonuclease digestion, 170, 288 purines and pyrimidines, free radicalinduced products, GC/MS, 193, 842 radical-damaged, hydrolysis, 186, 512 radical-mediated damage in presence of oxygen, 186, 511 recombinant, yeast clone selection, 194, 241 for gene isolation with hgtl I system, 194, 232 for reconstitution of nucleosome cores, length requirements, 170, 577 regulatory sites, consensus patterns, 183, 211 replication forks chromatin organization at, visualization, 170, 290 DNA sequence at, relationship to chromatin structure, 170, 312 first nucleosome on, distance to ends of nascent DNA, 170, 308 Okazaki fragment release, 170, 311 sequence relationship to chromatin structure, 170, 312

DNA simian virus 40, chromatin structure at, 170, 291 -retinoic acid receptor interactions in vitro, 189, 263 ribosomal, transcription initiation sites, mapping, 180, 103 - R N A crosslinks formation, psoralen method, 170, 157 purification, 170, 157 - R N A hybridization for primer extension analysis of RNA, 180, 361 principles and reaction conditions. 180, 242 in pulse labeling of hnRNA, 180, 92 for RNA characterization by nuclease S~ analysis, 180, 340 for RNA mapping studies, 180, 247 second-dimensional electrophoretic resolution, 170, 129 sequencing bands, colorimetric detection, 184, 612 single-stranded hybridization probes, 168,402 - R N A heteroduplexes, structural interpretation, 180, 259 uncrosslinked, depletion from crosslinked samples, 170, 395 uracil-substituted, isolation from phage, 185,606 specific sequences, detection, 170, 129 specific structures and modified bases, immunodetection, 170, 248 staining for flow cytometry, 171, 579 structure, ~zp NMR analysis, 177, 310 SV40 chromosomal, replication in t~itro, 170, 48 synthesis assay, retinoid modulation of phorbol ester effects in skin, 190, 348 inhibition by amiloride and analogs, 191, 748 synthetic, in heterologous gene expression, 185, 6 target sequences immobilized on nitrocellulose, detection, 184, 604 templates for large-scale RNA production, 181. 560 small RNA synthesis, 180, 52

DNA transfected, amplification strategies, 185, 558 transfer to mammalian cells, 185, 559 unwinding, fluorometric analysis: strand breaks and repair in mammalian cells, 186, 550 yeast alteration by transformation with synthetic oligonucleotides, 194, 362 chromosomal bacterial transformation, 194, 323 genetic and physical maps, 194, 827 nomenclature, 194, 8 partially digested, size fractionation, 194, 220 preparation, 194, 322 purification, 194, 817 complementing, determination of genetic locus, 194, 227 direct sequencing by primer extension, 194, 550 footprint analysis by primer extension, 194, 557 fractionation by CsCI gradients, 194, 174 gene mapping, 194, 38 high-molecular-weight, preparation, 194, 218 isolation, 194, 169, 552 miniprep for shuttling plasmids, 185, 293 mitochondrial behavior in crosses, 194, 154 cloning in bacterial vectors, 194, 159 isolation, 194, 158 mutations, sequence analysis, 194, 160 restriction analysis, 194, 159 mutagenesis methods, 194, 273 and plasmid shuffling in vitro, 194, 302 shuttle, with bacterial transposons, 194, 329 with Ty transposon, 194, 342 positional mapping of genes, 194, 57 premeiotic synthesis, 194, 97 repeated sequences, correlation with

66 bands in restriction spectra, 194, 175 secretory proteins, transcription in vitro, 194, 675 template for reverse biochemical protein synthesis, design and preparation, 194, 523 transcription in vitro, 194, 524, 545 yeast artificial chromosome, clone libraries preparation, 194, 251 screening, 194, 265 storage, 194, 265 Z form affinity chromatography, 184, 328 occurrence in genome, 170, 248 preparation, 184, 332 DNA-directed RNA polymerase, see RNA polymerase DNA polymerase T7, in yeast DNA sequencing, 194, 553 Taq, in yeast DNA sequencing, 194, 556 DNA polymerase I in 3' labeling of DNA primers, 180, 358 purification from Escherichia coli, historical account, 182, 783 DNase I, see Deoxyribonuclease I DNase II, see Deoxyribonuclease II Dolichos biflorus

vegetative tissue, lectin purification, 179, 344 Dominance tests for fusion ability in yeast cell and nuclear mutants, 194, 780 Dopamine assay, 168, 435 Dose-response curves peptides, testing by hippocampal brain slice recording, 168, 129 relative, testing for insufficient hepatic stores of vitamin A, 190, 249 Dot-blot assays antibody detection, evaluation, 182, 668 anti-idiotypic antibodies in hybridoma supernatants, 178, 86 carbohydrate receptors, 179, ll8 oxytocin and vasopressin mRNA in brain, 168, 398 proteins, 184, 417

67 RNA splicing in Escherichia coli, 181, 530 Doxorubicin -anti-idiotypic conjugate, in treatment of murine B-cell tumors pharmacological activity, 178, 374 preparation, 178, 371 Drosophila melanogaster, see Fruit fly Drugs antitumor, see Antitumor drugs binding to isolated nephron segments, microassay, 191, 315 efficacy testing with ascites tumors, 171, 601 induced oxidant stress, assay in perfused organs, 186, 759 induced oxygen radical generation, assay in vitro, 186, 621 DT-diaphorase, see NAD(P)H dehydrogenase (quinone) DTPA, see Pentetic acid DTSSP crosslinking of anion-exchange channels in intact erythrocytes, 172, 625 membrane-impermeant crosslinking reagent, 172, 619 synthesis, 172,622 Durapatite chromatographic columns guidelines for use, 182, 333 protein adsorption, 182, 330 protein elution, 182, 332 RNA adsorption, 180, 29 Dwell times for selected-ion monitoring, selection, 193, 98 Dyes, see also specific dyes anionic, in measurement of potential in cells and vesicles, 191, 478 cationic, in measurement of potential in cells and vesicles, 191, 476 fluorescent in assay of intracellular ion activities and membrane transport, 192, 38 Ca2+-sensitive, in analysis of membrane K + conductance, 192, 309 in cell type identification and analysis in isolated nephron segments, 191, 253

Edetic acid excitation ratios, measurement, 192. 39 loading of parietal cells general methods, 192, 39 toxic effects, 192, 45 in measurement of pH of intracellular compartments, 172, 85 membrane-permeant compartmentation, 192, 48 incomplete hydrolysis, 192, 46 toxic effects, 192, 45 impermeant, in measurement of pH of intracellular compartments, 172, 89 lethality, 186, 642, 645 mediated photooxidation, in generation of oxygen radicals in vitro methods, 186, 637 in vivo methods, 186, 641 for membrane potential measurement, 172, 104 reactive, immobilized for affinity chromatography of proteins, 182, 343 columns with, preparation, 182, 350 regeneration and storage, 182, 349 DYNPRO program extension for detection of pseudoknots and hairpins in RNA, 183, 313

E Ear mouse, phorbol ester-induced edema. measurement, 190, 348 Earthworm isolation of ceramide glycanase. 179. 481 Edema ear, phorbol ester-induced, measurement, 190, 348 salivary gland, development during perfusion, 192, 10 Edetic acid in dissociation of gastric mucosa into individual cells, 171, 446 effect on superoxide dismutase mimic prepared from MnO2 and desferrioxamine, 186, 246 and heat, abolition of interference with CAT assay, 168, 695

Edetic acid inhibition of enzymatic addition of guanylate to tRNA n~s, 181, 460 -iron chelates, role in hydroxyl radical production by Fenton reaction, 186, 15 in release of cells from culture for flow cytometry, 171, 567 in spin trapping of free radicals, 186, 131 Edman degradation manual, and liquid secondary MS, in amino acid sequencing, 193, 491 in N-terminal amino acid sequencing, 182, 602 EDTA, see Edetic acid Efflux coefficients for proximal tubule membranes, determination, 191, 212 Eggs, see also Oocytes Xenopus laevis

collection, 170, 613 dejellying, 170, 615 extracts assembly of regularly spaced nucleosomes, 170, 580 comparison, 181, 280 preparation, 181, 274 heat supernatant, preparation, 170, 616 purification of nucleoplasmin, 170, 612 Eicosanoid receptors detection in mesangial cells, 187, 546 Eicosanoids action, bioassay, 187, 605 biochemistry in cultured glomerular mesangial cells, 187, 544 chirality analysis by reaction with lipoxygenase, 187, 194 HPLC analysis, 187, 187 nomenclature, 187, 187 conversion to diastereomer, 187, 192 enzyme immunoassay with acetylcholinesterase, 187, 24 human lung parenchyma producing, preparation, 187, 621 metabolism in isolated perfused rat lung, 187, 608 related enzymes, nomenclature, 187, 3 production by endothelial cells assay, 187, 531

68 influencing factors ex vivo~ 187, 533 related interactions of platelets, endothelial cells, and neutrophils, 187, 585 release and effects in isolated coronaryperfused mammalian heart, 187, 610 role in pulmonary vasoregulation, 187, 607 Ektacytometry red blood cells, 173, 35 Elastic modulus cell wall, determination, 174, 353 ecological significance, 174, 210 plant cells and tissues, 174, 205 Electrical resistance airway epithelial cells, 192, 569 in determination of paracellular shunt conductance in epithelia, 191, 4 determination from transmucosal potential difference, 171, 611 intracellular, measurement on intact gastric mucosa, 192, 86 isolated gastric glands, 192, 90 isolated gastric mucosa cells, 192, 91 parietal cell cultures, 192, 91 lumen-tubular cell pathway and parallel pathways, 171, 624 membrane in epithelium, 171, 719 in gall bladder epithelial cells, 192, 657 paracellular shunt, determination, 191, 16 transepithelial, determination for gall bladder epithelium, 192, 657 gastric mucosa, 192, 84 isolated peffused renal tubules, 191, 291 mammalian urinary bladder, 192, 642 renal cell tissue culture, 191, 435 tubular structures, 192, 23 transmucosal, change in transition from resting to secretory state, mechanism, 171, 624 Electric current density, extracellular, in epithelia, electrophysiological scanning, 171, 793 electrogenic pump, patch clamp measurements, 174, 329 injection into giant algal cells, limitations, 174, 421 in intestinal secretory cell membrane

69 effect of intracellular Ca 2+, 192, 316 measurement, 192, 311 macroscopic, in gastric mucosa, measurement, 192, 83 noise measurement, 171, 285 relaxation after voltage jump, 171, 277 short circuit in determination of paracellular shunt conductance, 191, 4 across epithelial sheets, effects of furosemide-related diuretics, 191, 791 equivalent, in isolated perfused renal tubules, 191, 294 in large intestine in vitro measurements, 192, 476 in vivo measurements, 192, 468 in lepidopteran midgut, agreement with net K ÷ flux, 192, 607 in renal cell tissue culture, measurement, 191, 435 in small tubular structures, determination, 192, 15 in urinary bladder, measurement in mammals, 192, 644 toad, 192, 684 through single channels, patch clamp measurements, 174, 325 step, response of transmucosal potential difference, 171, 607 -voltage relation, in determination of paracellular shunt resistance, 191, 16 whole-cell, patch clamp measurements, 174, 329 Electric field mediated DNA transfection of mammalian cells, 185, 531 permeabilization of mammalian cells to macromolecules, 171, 868 Electric potential in analysis of ion and nonelectrolyte permeation, 171, 703 biionic, in renal cell tissue culture, measurement, 191, 435 in cells and vesicles, measurement, 191, 476 constant accelerating, linked MS scans with, 193, 48 -current relation, in determination of

Electrochemical gradients paracellular shunt resistance, 191. 16 divider ratio, in determination of paracellular shunt conductance, 191, 9 interfacial, in biomembranes, lipid coumarin dye as probe, 171, 376 intracellular, measurement on bovine corneal endothelial cells, 192. 574 intact gastric mucosa, 192, 86 isolated gastric glands, 192, 90 isolated gastric mucosa cells, 192, 91 isolated perfused renal tubules 191, 295 parietal cell cultures, 192, 91 intrasymplast, measurement, 174, 287 intravessel, measurement, 174, 286 jump, current relaxation after, in analysis of ion carriers in planar bilayers. 171, 277 junction, in analysis of ionic permeation mechanisms, 171,688 relaxation after charge pulse, in analysis of ion carriers in planar bilayers. 171, 279 streaming, in analysis of ion and nonelectrolyte permeation, 171. 707 transepithelial in avian renal tubules, properties. 191. 225 measurement in gall bladder epithelium, 192,657 gastric mucosa, 192, 83 insect Malpighian tubules, 192,629 large intestine, 192, 461,469 mammalian urinary bladder, 192, 644 perfused Malpighian tubule in vitro: assay of diuretic hormone. 191. 178 proximal tubules from reptile. 191. 207 transmucosal, response to step currents. 171, 607 trans-root, measurement, 174, 286 Electroblotting proteins, 182,604, 680 Electrochemical gradients of defined magnitude, creation in intestinal epithelial cells, 192, 337

Electrochemical gradients protons across plasma membrane, determination, 174, 632 in vesicles, organelles, and prokaryotic ceils, measurement, 172, 63 Electrodes ion-selective, in determination of lipophilic ion distribution, 172, 67 net ion fluxes in C h a r a cells, 174, 460 macro-, s e e Macroelectrodes micro-, s e e Microelectrodes oxygen, s e e Oxygen electrodes pH, in measurement of plant membrane potential, 174, 298 reference, for measurement of ionic permeation properties, 171, 685 scanning, in localization of epithelial transport pathways, 171, 792 Electrodiffusion simple, ions, 171, 69 Electrogenicity and electroneutrality, in cation-anion transport, comparison, 173, 287 Electrolytes effects on dsDNA thermal stability, 180, 241 transport intestinal cell lines as models, 192, 354 in red blood cells, light scattering studies, 173, 272 in tracheal epithelium, analysis, 192, 555 transepithelial in renal tubules, structural basis, 191, 266 Ussing chamber studies, 192, 361 Electron capture in chemical ionization, 193, 17 Electroneutrality in alkali metal/proton exchange, assessment, 173, 336 and electrogenicity, in cation-anion transport, comparison, 173, 287 Electronic particle analyzers in measurement of platelet aggregation in vitro, 169, 134 platelet counting with, 169, 165 Electronic sizing in detection of Na+/H ÷ exchange-induced cell swelling, 173, 779

70 Electron microscopy in analysis of phloem loading, 174, 297 avidin-biotin probes, 184, 415 avidin probes, 184, 413 chromatin, 170, 142 contrast enhancement, 180, 492 cytoplasmic organelles, 182, 223 dark-field images, comparison with bright-field, 169, 329 metal replication with tungsten, 169, 328 platelet adhesive proteins, 169, 326 DNA-RNA heteroduplexes, 180, 252 dsDNA, 180, 252 glucose transporter from human erythrocytes, 174, 44 in identification of polymerase I, II, and III genes, 180, 497 immuno-, s e e Immunoelectron microscopy in localization of biotinylated gonadotropin-releasing hormone, 184, 401 in mapping RNA genetic organization, 180, 239 Miller chromatin-spreading method, 180, 482 myosin ATPase site, 184, 409 nucleosomes arcs and cylinders from purified core particles, 170, 110 characterization as chromatin repeat unit, 170, 101 proteins, for molecular size determination and subunit detection, 182, 585 ribonucleoprotein complexes hnRNP after reconstitution, 181, 313 on nascent RNA, 180, 481 R-loops, 180, 252 scanning, glass beads after interaction with platelets in vitro, 169, 113 simian virus 40 minichromosomes, 170, 166 nucleosomes, 170, 176 thin-section, yeast cell preparation for, 194, 602 thrombospondin, 169, 266 transmission in evaluation of carbohydrate receptor binding by colloidal gold, 179, I 17 scanning, SV40 chromatin, 170, 175

71 visualization of intracellular protein trafficking, 184, 379 yeast nuclei, 194, 129 yeast nucleolus, 194, 717 Electron multiplier detectors in mass spectrometry, 193, 65 Electron paramagnetic resonance in measurement of electrostatic potential adjacent to membranes, 171, 358 free radical reductase in biological tissues, 186, 671 pH gradients, transmembrane potentials, and membrane dynamics, 172, 331 Electron probe X-ray microanalysis associated radiation damage and contamination, 172, 221 biological applications, 172, 223 cell composition, 171, 764 fluid secreted by Malpighian tubules, 191, 173 ion compartmentation in root cells and tissues, 174, 267 ions in rapidly frozen cells, 172, 203 principles, X-ray detectors, and calibration, 172,209 spatial resolution and sensitivity, 172, 219 spectral processing, 172, 212 validation, 172, 220 X-ray mapping, 172, 225 Electron spin resonance in analysis of human lung from cigarette smokers, 186, 665 lignin peroxidase, 188, 163 retinoid-phospholipid membrane interactions, 189, 383 in assay of biomembrane surface potential, 171, 373 free radical reductase in biological tissues, 186, 670 retinoid antioxidant activity, 190, 274 o-semiquinone radicals, 186, 333 tocopheroxyl radicals, 186, 197 Electron transport chain direct interaction with glucagon, 174, 125

Electrophysiology Electropermeabilization pancreatic acinar cells, 192, 289 Electrophoresis, see also specific techniques

in analysis of peptide processing in vivo, 168, 521 ascites containing monoclonal antibody, 178, 56 capped mRNA digests on paper, 180, 171 tree-flow apical and basolateral hepatocyte plasma membrane vesicles. 191. 836 in plasma membrane preparation, 172, 12 in purification of basolateral plasma membranes from renal cortex. 191, 457 lumenal and contralumenal membranes simultaneously, 191, 459, 466 in separation of cell populations. 171. 513 immunocompetent cells. 171, 517 malaria parasites, 171, 521 renal proximal and distal tubule cells, 171, 525 tumor cells, 171, 530 N-glycosylated invertase, mobility shifts. 194, 685 high-voltage, RNA oligonucleotide branch points, 180, 189 O-mannosylated chitinase, mobility shifts, 194, 687 in measurement of electrostatic potential adjacent to membranes, 171,346 in protein concentration and solute removal, 182, 82 RNA digests on cellulose acetate. 180, 141 thin-layer, and TLC, peptides, 182, 623 Electrophoretic transfer high-molecular-weight proteins for immunostaining, 172, 687 Electrophysiology in antibody mapping of sodium channel functional sites, 178, 728 opioid peptides. 168. 103

Electroporation Electroporation in gene transfection for chimeric antibody production, 178, 468 introduction of DNA into mammalian cells, 185, 531 in transformation of yeast, 194, 182 Electrospray sample application for plasma desorption MS, 193, 433 Electrostatic potential adjacent to membrane, measurement, 171, 342 Elemental analysis retinoic acid analogs, 189, 37 /3-Elimination mannoproteins, 185, 466 ELISA, see Enzyme-linked immunosorbent assay Elongation factors discovery, resolution, purification, and function, 182, 809 Elution proteins from electrophoretic gels, 182, 488 Elutriation gastric mucosa endocrine cells, 192, 179 major and minor cell types from complex tissues, 171,482 Elvax 40P implants, preparation, 187, 382 EMBL data library data acquisition, 183, 27 databases, 183, 23 data distribution, 183, 28 Embryo chicken cardiac cells, preparation, 173, 637 developed, cartilage pattern, 190, 206 local application of retinoic acid and congeners, 190, 202 targeted slow release of retinoids into, 190, 201 Drosophila, nuclei and homogenates, preparation, 170, 321 sea urchin, active nuclei, preparation, 181, 34 EMS, see Ethyl methanesulfonate Endocrine cells canine gastric mucosa isolation and primary culture, 192, 176

72 peptide release studies, 192, 184 transient permeabilization, analysis of inositol lipid metabolism, 168, 218 Endocytosis in cells of isolated nephron segments, analysis, 191, 255 proteins in proximal tubules, 191, 340 tracer studies, 191, 557 transporter removal in regulation of membrane transport, 172, 49 yeast, assays, 194, 697 Endo-/3-galactosidase releasing Gal(al---~3)Gal assay, 179, 496 properties, 179, 499 purification from Clostridium perfringens, 179, 498 Endoglycosidase H in analysis of yeast glycoproteins, 194, 694 Endoglycosylceramidase assay, 179, 489 properties, 179, 493 purification from R h o d o c o c c u s sp. G-742, 179, 490 Endo-al--~6-mannanase,see Mannan endo- 1,6-/3-mannosidase Endonucleases H O , in analysis of yeast chromosome structure, 194, 143 Endoplasmic reticulum hepatic, intactness measurement, 174, 58 rough, marker RNA assay, 182, 221 yeast enzyme marker assay, 194, 658 immunofluorescent detection, 194, 601 Endopolygalacturonase, see Polygalacturonase Endoproteinases protein digestion for MS analysis, 193, 363, 367 Endorphin receptors antibodies, production with complementary peptides, 168, 25 /3-Endorphins OmpF, secretion from Escherichia coli, 185, 180 Endosomes marker enzyme horseradish peroxidase, assay, 182, 217

73 pH measurement, 173,755 pH and proton pumping, assay in cell cultures, 174, 143 isolated organelles, 174, 151 Endothelial cells arachidonic acid metabolism, 187, 531 aspirin treatment, 187, 588 bovine corneal, culture and transport studies, 192, 571 macrovascular, culture, 187, 526 eicosanoid production e x v i v o , influencing factors, 187, 533 glomerular, isolation and culture, 191, 148 hepatic, and Kupffer cells, separation, 190, 56 human primary culture, 187, 521 umbilical vein, prostaglandin Iz synthesis, 187, 539 labeling with [3H]arachidonate, 187, 532 monolayers, platelet interactions with, measurement, 169, 71 platelet-activating factor production, 187, 527,533 and platelets and neutrophils, eicosanoid interactions, 187, 585 porcine aortic cellular integrity measurement, 187, 563 interaction with neutrophils: leukotriene C4 synthesis, 187, 559 preparation, 187, 560 rat, culture preparation, 187, 400 role in free radical-mediated reperfusion injury, 186, 717 for studies of platelet-activating factor and arachidonate metabolites, 187, 521 Endothelium role in ischemia-reoxygenation injury, 186, 78 sub-, s e e Subendothelium Endotoxin induced circulatory shock, role of oxygen radicals, 186, 660 induced disseminated intravascular coagulation, analysis, 186, 657 Energy rules

Enzymatic reactions for computer prediction of RNA structure, 180, 264 Energy transfer nonradiative singlet-singlet, in measurement of nucleosome internal distances, 170, 208 in oxidative phosphorylation, nonequilibrium thermodynamics, 171,403 resonance, lipid probes in liposomes, microscopic study, 171,852 Enkephalin pre-pro-protein, s e e Preproenkephalin Enniatin A ion carrier in planar bilayers, relaxation and noise analyses. 171, 274 Enniatin B ion carrier in planar bilayers, relaxation and noise analyses, 171, 274 Entactin differentiation marker for embryonal carcinoma cells, 189, 345 Enterocytes isolation, 192, 397 membrane isolation, 192, 341 Enterotoxins E s c h e r i c h i a coli, release mechanism, 185, 174 Enzymatic assay aminopeptidases, 194, 450 avidin, biotin, and streptavidin, 184, 208 carboxypeptidase S, 194, 449 carboxypeptidase Y, 194, 448 catalytic antibodies, 178, 544 colorimetric, avidin and biotin, IlM, 217 coupled benzoate-CoA ligase, 188, 154 dichloromethane dehalogenase, 188, 355 glutathione in biological samples, 173, 525 and glutathione disulfide, 186, 758 metabolites in leaf cells and subcellular compartments, 174, 548 radioactive, s e e Radioenzymatic assay yeast protease A, 194, 444 yeast protease B, 194,447 yeast vacuolar proteases, 185, 380 Enzymatic reactions steady-state rate equations, generation, 171, 164

Enzyme immunoassay Enzyme immunoassay with acetylcholinesterase eicosanoids, 187, 24 leukotrienes C4 and Ea, 187, 82 validation with GC/MS, 187, 33 antibody detection, evaluation, 182, 668 antisubstance P anti-idiotypic antibodies, 178, 285 avidin-biotin system for, 184, 469 competitive additive, carcinoembryonic antigen, 184, 514 idiotype-anti-idiotype, adenosine deaminase-binding protein, 178, 171 solid-phase, epitope specificities, 184, 509 solid-phase, for screening hybridomas, 178, 54 Enzyme-linked immunosorbent assay in antibody mapping of sodium channel functional sites, 178, 727 antihistone antibodies, 170, 258 anti-idiotypic antibodies to opioid receptors, 178, 248 purified, 178, 162 anti-PADGEM protein antibodies, 169, 314 antisera analysis in preparation of immunological probes for nucleosomes, 170, 220 with biotin-streptavidin, herpes simplex virus, 184, 547 with biotinylated protein A, description, 184, 497 catalytic antibodies, 178, 542 competitive, high-molecular-weight kininogen, 169, 281 enhancement with biotin-avidin, 184, 537 histidine-rich glycoprotein from platelets, 169, 269 human anticarbohydrate antibodies, 179, 297 inhibition assay interspecies cross-reactive idiotopes, 178, 384 paratope-related idiotopes, 178, 383 interphotoreceptor retinoid-binding protein, 189, 219

74 monoclonal anti-Id antibodies to poliovirus, canine parvovirus, and rabies virus, 178, 382 with peptide antigens, for distinguishing HIV-1 and HIV-2 infections, 178, 705 for screening haptoglobin monoclonal antibodies, 182, 712 monoclonal antibodies, 182, 672 transfectomas producing chimeric antibodies, 178, 469 site-specific antipeptide antibodies to rhinovirus receptor, 178, 683 Enzymes, see also specific e n z y m e s activity units, definition, 182, 42 antibodies mimicking, see Antibodies, catalytic assays anatomy, 182, 38 classification, 182, 40 design, 182, 43 biotinylated, complexes with avidin and streptavidin, preparation, 184, 181 biotinylation, 184, 474 coupling to avidin or streptavidin, 184, 475 for DNA and RNA hydrolysis, 193, 785 eicosanoid metabolism-related, nomenclature, 187, 3 -ligand interactions, NOE studies, 177, 333 multienzyme complexes, see Multienzyme complexes organellar marker, assay considerations and bookkeeping, 182, 211 proteolytic, effects on vacuolar arginine permease, 174, 515 purification by heat and pH denaturative precipitation, 182, 298 significance, 179, 1 specific activity, definition, 182, 42 -streptavidin complexes, in detection of antigens on Western blots, 184, 437 Epidermal growth factor human, export from Escherichia coli, 185, 183 Epidermal growth factor receptors phosphorylation

75 effect of ganglioside Gin, 179, 525 inhibition by lyso-G~t3, 179, 531 stimulation by de-N-acetyl-Gm, 179, 531 Epidermis, see also Skin biopsied, free radical reduction at surface, 186, 674 detection of newly formed 3,4-didehydro[3H]retinol, 190,212 human, lipid composition and effects of retinoic acid, 190, 39 peeled, preparation of guard cell protoplasts, 174, 314 radioactive retinyl and 3,4-didehydroretinyl esters, analysis, 190, 213 reconstructed, lipid extraction, 190, 32 renewal, retinoid effects, 190, 76 Epididymis purification of lysosomal c~-D-mannosidase, 179, 501 Epinephrine assay, 168, 435 effect on citrated platelet-rich plasma, 169, 131 Epithelial cells airway electrolyte transport, analysis, 192, 555 electrophysiology, 192, 557 isolation, 192, 552 tight junction formation and transepithelial resistance, 192, 569 transformation with persistence of cystic fibrosis phenotypes, 192, 565 Ca2+-transporting, Ca 2+ as messenger in, assessment, 191, 674 chloride channels blockers, 191, 802 purification and reconstitution, 191, 572 compositional analysis, 171,750, 758 differentiation analysis with porous-bottom culture dishes, 171, 736 as measure of vitamin A status in human populations, 190,247 role of retinoids, 190, 19 glomerular, isolation and culture, 191, 150

Epithelial cells intestinal ATP-depleted, in analysis of transport functions, 192, 335 basolateral membrane Ca 2+ transport, 192,448 electrochemical gradients of defined magnitude, creation, 192, 337 intracellular Na + and K +, assay, 192, 337 membrane potentials of defined magnitude, creation, 192, 337 measurement, 171, 772; 192, 338 c~-methylglucoside accumulation, 192, 329 Na ÷ gradients of defined magnitude. creation, 192, 336 Na+-independent sugar transport, 192, 334 transport functions, evaluation, 192. 324 villous, isolation, 192, 324 membranes ionic selectivity, 171, 727 resistance properties, 171,719 messenger generation and action, separation, 191, 672 renal dispersions, preparation, 171, 586 fusion, 192, 715 isolated, characterization, 171, 588 phosphate transport in established cell lines, 191, 494 retinal pigment cellular retinoid extraction and analysis, 190, 13 retinoid metabolism in vitro, 190, 16 all-trans-[3H]retinol supply in vitro. 190, 15 short-term incubation for analysis of visual cycle, 190, 3 viable, isolation, 190, 4 stimulus-secretion coupling, 191, 661 tracheal differentiation, role of retinoids, 190, 91 electrogenic and electroneutral ion transporter regulation, 192, 549 keratin gene expression during vitamin A deficiency, 190, 95

Epithelial cells keratin synthesis, role of retinoids, 190, 91 morphological alterations, 190, 91 preparation, 190, 94 transport analysis with porous-bottom culture dishes, 171, 736 overview, 191, 1 pathway localization, 171, 792 volume regulation, experimental approaches, 171, 744 Epithelium direct current electrical measurements, 171, 6O7 exocrine activation, patch-clamp studies, 171, 663 extracellular current density, electrophysiological scanning, 171, 793 gall bladder, ion and water transport in Necturus, 192, 650 ionic permeation mechanisms, 171, 678 large intestine electrophysiology in vitro, 192, 468 layers, electrical measurements, 192, 480 resistivity in vivo, 192, 466 layer preparation on permeant supports, 191, 432 leaky, impedance analysis, 171, 642 morphology, in analysis of vitamin A status in human populations, 190, 247 paracellular shunt conductance, determination, 191, 4 renal analysis in situ, 191, 273 isolation with cell-specific monoclonal antibodies, 171, 581 mammalian, preparation for study of cell volume regulation, 171, 783 quantitative evaluation, 191, 281 segmental transport functions in vitro, 191, 418 retinal pigment cellular retinaldehyde-bindingprotein, binding of 1l-cis-retinaldehyde, 189, 369 retinoid isomerase assay, 189, 503 sheets preparation for study of cell volume regulation, 171, 789

76 short-circuit current, effects of furosemide-related diuretics, 191, 791 subepithelial puncture leaks, effect on impedance analysis, 171, 660 tight equivalent circuit models, 171, 638 impedance analysis, 171, 628 tubular, elasmobranchs function, 191, 182 isolation, 191, 183 urinary bladder electrical recording, 192, 641 H + transport regulation, 172, 54 physiological studies, 192, 632 transport properties, 192, 648 viability during transepithelial studies, 192, 647 Epitopes affinities, solution-phase assay, 184, 518 bacterial network, 178, 12 B-cell computer prediction, 178, 586 identification in hepatitis B virus proteins, 178, 639 glycanic schistosomal, mimicry by antiId antibodies, 178, 392 HIV, for preparation of synthetic peptide antigens, 178, 708 identification by hydrophilicity plotting, 178, 571 mapping with protein footprinting, 178, 746 nucleosome, exposure, 170, 234 peptide B-cell, addition of Th-cell epitopes, 178, 661 with enhanced immunogenicity, vaccines based on, 178, 659 presentation on hepatitis B core protein, 178, 669 size-exclusion HPLC analysis, 178, 107 protein size-exclusion HPLC analysis, 178, 107 tagging in yeast, 194, 508 specificities, solution-phase assay, 184, 518 T-cell identification and use in vaccine construction, 178, 611

77 mapping by k-tuple frequency analysis, 183, 237 structure, amphipathic helix hypothesis, 178, 616 synthetic, for inducing antibodies to hepatitis B components, 178, 635 Th-cell, addition to B-cell epitope peptides, 178, 661 viral network, 178, 8 EPMA, s e e Electron probe X-ray microanalysis Epon in embedding of artery segments, 169, 62 Epoxidation arachidonic acid, 187, 360 methyl arachidonate, 187, 361 Epoxide hydrolase hepatic purification from cytosol, 187, 328 subcellular distribution, 187, 327 hydrolysis of 2,2-dimethyloxirane, mechanistic studies, 177, 387 kinetics, 187, 331 substrate specificity, 187, 327 Epoxy acids nomenclature, 187, 1 Epoxyeicosatrienoic acids and heteroatom analogs, synthesis, 187, 357 I802-1abeled, preparation, 187, 184 production by hepatic microsomes, assay, 187, 182 stable isotope-dilution MS assay, 187, 175 EPR, s e e Electron paramagnetic resonance Epstein-Barr virus vectors for heterologous gene expression in mammalian cells, 185,493 Equilibrium constants enzyme-bound reactants and products, determination, 177, 358 Ergothioneine antioxidant activity, 186, 310 applications, 186, 316 assay, 186, 318 as chelating agent, 186, 314 defensive function, 186, 312 distribution, 186, 314 pharmacology, 186, 316 properties, 186, 310

Erythrocytes Erythritol permeability of phospholipid membranes, effects of retinoids, 189, 379 Erythrocytes amino acid transport effect of cell maturation, 173, 156 flux measurements, 173, 130 inhibitor studies, 173, 147 kinetics, 173, 138, 149 role of ions, 173, 142 thermodynamic effects, 173, 156 amino acid transporter, identification. 173, 122 A m p h i u m a , alkali-metal exchange identification criteria, 173, 334 net flux studies, 173, 331 anion transport assay and analysis, 173, 54 flow tube assay, 173, 160 steady-state rate equations, generation, 171, 177 ATP-depleted, A23187-induced Ca :+ permeabilization, 173, 110 ATP levels, control, 173, 105 cation fluxes: Na*,K + pump, 173, 80 cell content assay, 173, 60 cell-free extracts, preparation, 186, 500 chelator-containing, Ca 2~ loading. 173. 109 chicken, nuclei and homogenates, preparation, 170, 318 isolation, 170,434 chloride loading using nystatin, 173, 438 conformation, relationship to membrane chemical equilibrium, 173, 12 coupling to avidin, 184, 317 cytoskeleton shells, preparation and coating with lipids, 173, 394 deformation direct measurement, 173, 17 ektacytometric index, 173, 45 micromechanical test system, 173, 14 oxygen effects, 173, 51 properties, 173, 3 ektacytometry, 173, 35 electrolyte transport, light scattering studies, 173, 272 fragmentation, ektacytometric analysis. 173.49

Erythrocytes ghosts hemolytic holes, analysis, 173, 356 membranes, preparation, 173, 495 preparation, 173, 357, 514 proteolysis, 173, 428 resealed exofacial modification, 173, 469 with varying CI~, preparation, 173, 439 resealing, 173, 357 stripped, solubilization, 173, 500 transport assay anions, nonelectrolytes, and water, 173, 160 fluorescence methods, 172, 129 white, resealable, preparation, 173, 346 glucose transport, characterization, 171, 12 human Ca2+ content and transport, 173, 101 equilibrium exchange of 3-methoxyglucose, 191, 771 intact, protein ligand association with band 3,173, 518 interaction with neutrophils: leukotriene B4 synthesis, 187, 553 isolation, 187, 555 Li÷/Na ÷ exchange, assay, 173, 184 lithium efflux, assay, 173, 187 lithium loading, 173, 186 membrane potential, measurement with dyes, 172, 102 membranes cytoplasmic surface, protein associations with band 3,173, 513 cytoskeletal protein, removal, 174, 41 isolation, 173, 414 lipid core, photochemical labeling, 172, 659 perturbation and photolabeling by azides, 191, 771 preparation, 174, 40 microcapsule preparation, 171, 217 22Na÷ efflux, assay, 173, 188 Na+/H + exchange assay, 173, 181 kinetics, 173, 176 Na+/Li ÷ exchange, assay, 173, 184

78 Na ÷ loading, 173, 187 Na+/Na ÷ exchange, assay, 173, 184 pH modification, 173, 178 purification of Cu-Zn superoxide dismutases, 179, 573 glucose transporter, 174, 39 glycolipids, 179, 41 single-channel currents, recording, 173, 112 sugar transport, inhibition, 191, 773 incubation with phospholipases, 173, 227 intact crosslinking of anion-exchange channels, 172, 625 Heinz body formation, 186, 272 intracellular ATP levels, modification, 173, 86 intracellular Ca2÷, assay and control, 173, 100 intracellular cation content, alteration, 173, 81 [~4C]phosphatidylcholine introduction, 173, 226 protein ligand association with band 3, 173, 518 lipid analysis, 173, 227 lysates, preparation, 186, 227 maturation, effect on amino acid transport, 173, 156 mediated DNA transfection of mammalian cells, 185, 534 membrane potential, measurement, 173, 94 membranes anion-exchange protein isolation, reconstitution, and orientation, 173, 410 bulk D20 diffusion across, 173, 199 chemical equilibrium, relationship to cell conformation, 173, 12 cytoplasmic surface, protein associations with band 3, 173, 513 failure mechanisms, 173, 29 inside-out vesicles Na+,K+-pump stoichiometry and coupling, 173, 377 preparation and properties, 173, 368 intact, macromolecule crosslinking, 184, 158

79 leaflets, p4C]phosphatidylcholine equilibration between, 173, 227 material properties, 173, 8 mechanical properties, 173, 17 nucleoside transport, 173, 250 osmotic water flow across, 173, 204 reassociation of protein ligands with, 173, 516 skeleton proteins, preparation, 173, 380 water diffusion across, 173, 192 monocarboxylate transport, kinetics and modification, 173, 300 Na+-Ca 2. and Na+-H + exchange, analysis, 173, 292 nonelectrolyte transport light scattering studies, 173, 268 measurement, 173, 160 ovine antibody response in mouse, in assay of retinoid immunomodulatory activity, 190, 256 purification of glycolipids, 179, 39 permeabilization to Ca 2+ by A23187, 173, 107 pH, intracellular, cells varying in, preparation, 173, 57 pigment free, transport assay, 172, 129 proteolysis, 173, 428 radioactive anion-containing, preparation, 173, 58 reconstitution, preparation of phospholipid membrane-coated cytoskeletons, 173, 392 structural properties, 173, 3 sugar transport, analysis biochemical methods, 173, 234, 249 optical methods involving osmotic swelling or shrinking, 173,235 radioactive methods, 173, 243 suspensions, reflection coefficient measurement, 173,213 transbilayer mobility of phosphatidylcholine, 173, 223 transport assay fluorescence methods, 172, 128 water, 173, 160 Erythropoietin recombinant human, characterization, 193, 524

Escherichia coli Erythrosomes characterization, 173, 395 preparation, 173, 394 Escherichia coli antibiotic action, resistance, and working concentrations, 185, 22 calcium-shocked, preparation and transformation, 194, 326 cell-free extracts, preparation, 186, 500 clpA mutants for minimization of proteolysis, 185, 126 colonies, superoxide dismutase assay. 186, 240 conjugation with yeast in transmission of plasmid DNA, 194, 187 disordered human proteins in, refolding. 185, 187 dut ung , phage growth in, 185, 605 dye-mediated lethality, 186, 642. 645 engineering to secrete heterologous gene products, 185, 166 Eo -711, promoter recognition and mRNA initiation, 185, 37 export of eukaryotic proteins, 185, 180 expressed antibody fragments, purification, 178, 512 expressed proteins in aggregates, solubilization, 182, 264 expression of avidin, 184, 75 functional antibody Fv and Fab fragments, 178, 497 heterologous proteins, 185, 11 mammalian cellular retinol-binding proteins, 189, 506 recombinant proteins, gene fusions for, 185, 129 expression systems, overview, 185, 5 extracts and subcellular fractions, preparation, 181, 360 htpR mutants for minimization of proteolysis, 185, 125 lactose transport, characterization. 171. 12 Ion mutants filamentation in, 185, 124 for minimization of proteolysis, 185. 120 phenotypic confirmation, 185, 124

E s c h e r i c h i a coli secondary phenotypes, elimination, 185, 121 mechanical lysis, 182, 152 mRNA translatability, influencing sequences in ribosome binding sites, 185, 103 oxygen radical production by photosensitization, 186, 635 pATH-transformed, Trl0 starvationinduced hybrid protein expression, 194, 486 pATH vectors, construction of trpE fusion genes in yeast, 194, 477 -phage hgtl I system, in isolation of yeast genes, 194, 230 plasmid vectors, design and construction, 185, 14 production of engineered antibody fragments, 178, 477 protease-defective, in enhancement of protein product stability, 182, 106 proteins in degradation, assay, 186, 496 overproduction, 182, 93 targeting pathways, 185, 169 proteolysis, minimization by genetic methods, 185, 119 protoplasts fusion, 185, 565 preparation, 185, 564 purification of DNA polymerase I, historical account, 182, 783 methionine aminopeptidase, 185, 407 polyadenylate-binding proteins, 181, 342 RNase III, 181, 192 RNase BN, 181, 431 RNase D, 181, 424 RNase P, 181, 569 RNase T, 181, 427 tRNA nucleotidyltransferase, 181, 436 yeast tRNA ligase, 181, 467 recovery of shuttle plasmids from yeast into, 194, 319 ribosomes, direction to single mRNA species, 185, 103 RNA splicing, genetic and molecular analyses, 181, 521

80 with rnpA-carrying plasmids, purification of C5 protein, 181,573 with rnpB-carrying plasmids, preparation of M1 RNA, 181, 578 rRNA maturation, assays, 181, 355 secretion of eukaryotic proteins, 185, 180, 185 heterologous proteins, 182, 132 prokaryotic exoproteins, 185, 177 strain selection for efficient expression system, 185, 33 superoxide dismutase deficient mutants, elevated rnutagenesis, 186, 646 induction by synthetic dyes, 186, 641, 644 transformation with yeast DNA, 194, 322 translation enhancement with two-cistron expression system, 185, 94 foreign genes, enhancing vector, 185, 115 high-level initiation, 185, 89 transposons for shuttle mutagenic manipulations in yeast, 194, 329 T7 RNA polymerase-directed expression of cloned genes, 185, 60 tRNA processing, genetics, 181, 395 trp promoter, in direct expression of proteins, 185, 54 yeast genes lethal to, trouble-shooting, 194, 229 -yeast shuttle vectors, in generation of yeast mutants from cloned genes, 194, 312 ESR, see Electron spin resonance Esterification retinol by acyl-CoA-dependent pathway, 189, 442, 446 retinol by lecithin-retinol acyltransferase, 189, 450 trans-, see Transesterification Estrogen antioxidant activity, 186, 330 radioimmunoassay, in granulosa cell aromatase bioassay of FSH, 168, 421

81 Estrogen receptors -ligand complexes, assay, 184, 297 purification by avidin-biotin affinity chromatography, 184, 273 with biotinylestradiol, 184, 292 Ethanol lipid solution injection, 171, 204 precipitation of proteins, 182, 296 Ether in testing for yeast sporulation, 194, 146 Ethidium bromide fluorescence, in assessment of parietal cell permeabilization, 192, 119 induction of yeast rho ° and rho strains, 194, 151 Ethylenediamine modification of DNA, 184, 602 Ethylene glycol bis(2-aminoethyl)ether N,N,N'.N'-tetraacetic acid Ca `,+ buffer, calculation and preparation, 172. 256 Ethyl hydroperoxide chloride peroxidase-catalyzed decomposition, ~O.,detection during, 186, 133 N-Ethylmaleimide covalent modification of phosphate transporter, 173, 738 sensitive phosphate-phosphate exchange, assay, 173, 736 Ethyl methanesulfonate induction of mutagenesis in yeast, 194, 277 mutagenesis of Schizosaccharomyces pombe, 194, 805 N-Ethyl-N-nitrosourea in structural probing of snRNA, 180, 214, 221 Ethyl (E)-retinoate deuterated at carbons 4 and 18, preparation, 189, 16 Etretinate HPLC with automated column switching, 189, 150 intragastric infusion via implanted osmotic minipumps, 190, 444 and metabolites, separation, 189, 151 pharmacokinetics, 190, 292 therapeutic value, 190, 297

Exocytosis Eukaryotic cells adhesion to immobilized carbohydrates. 179, 542 attached to substratum, Mg 2" influx measurements, 173, 558 higher, extract preparation, 182, 194 Mg 2+ transport, assay, 173, 546 protein overexpression, overview, 182, 112 subcellular isotope distribution, determination, 173, 553 Evaporation reverse-phase, in formation of large unilamellar vesicles, 171, 206 multilamellar vesicles, 171. 200 Evolution. see also Phylogenetic analysis genes, estimation, effect of base composition, 183, 570 human, and DNA sequence branching. 183, 550 nucleosomes, analysis with immunological probes, 170, 229 tree construction from aligned homologous sequences, 183. 601 Exchange rates computer calculation, 177, 458 ligands with enzymes, NOE study, 177. 338 and relaxation, related computer programs, 176, 480 Excimers formation in pyrene-labeled core nucleosomes, 170, 205 Excitation pulsed, frequency-selective, as selective relaxation method, 176, 186 selective, in solvent suppression for NMR, 176, 69 Excitation spectra fluorescence, 4-heptadeeyl-7-hydroxycoumarin, 171, 383 Exclusion chromatography histone subunits, 170,451 Exocytosis in cells of isolated nephron segments, analysis, 191, 257 fluorescent dextran, 172, 56 lipid mediators by bone marrow-derived mast cells, 187, 516

Exocytosis single-cell assay, 172, 57 transporter insertion in regulation of membrane transport, 172, 49 Exo-c~l---~2-mannosidase purification from Aspergillus phoenicis, 185, 463 Exonuclease III in analysis of nucleosome positions, 170, 639 in mapping of hypersensitive sites in chromatin, 170, 275 Exoproteins fusion-mediated implantation into plasma membranes, 171, 846 prokaryotic, secretion in Escherichia coli, 185, 177 Extinction coefficients protein of unknown concentration, 182, 56 solubilized protein at 280 nm, 172, 524 superoxide dismutase mimic prepared from MnO2 and desferrioxamine, 186, 244 Extracellular matrix modulation by retinoids: control of collagenase expression, 190, 175 subendothelial coated surfaces, preparation, 169, 79 components, degradation by platelets, 169, 85 megakaryocyte interaction with, 169, 87 platelet reactivity with, assay on coated dishes, 169, 82 in perfusion system, 169, 83 sulfate-labeled, preparation, 169, 86 vascular wall cells, preparation, 169, 49 Extraction arachidonic acid metabolites, 187, 67 automated on-line, lipoxygenase products of arachidonic acid, 187, 98 crosslinked ribonucleoproteins from UVirradiated HeLa cells, 180, 414 cytochrome P450 arachidonate metabolites, 187, 376 2,4-dinitrophenylhydrazine, filter paper method, 186, 473 direct, in semiquantitative analysis of retinoids, 190, 172 DNA-protein complexes with phenolchloroform, 170, 394

82 glucose transporter from brush border membranes, 192, 440 HI histones from core histones, 170, 526 high-molecular-weight RNA from yeast, 194, 399 histone subunits from nucleoprotein, 170, 450 4-hydroxyalkenals from tissues, 186, 402 inositol phospholipids from platelets, 169, 421 leukotriene A4 transformation products, 187, 325 lipid peroxides from tissues, 186, 389 lipids from human platelets, 187, 592 pancreatic tissue, 191, 686 reconstructed epidermis, 190, 32 lipoxins, 187, 169 mannoprotein from yeast mnn mutants, 185, 455 neuroendocrine peptides from milk, 168, 495 phospholipid hydroperoxides, 186, 453 phospholipids from pancreatic tissue, 191, 686 tissue, 187, 195 platelet-activating factor, 187, 153 polysomes with salt, 181, 205 proLH-RH from preoptic-hypothalamic tissue, 168, 527 protein kinase C from pituitary cells, 168, 289 proteins from ribosomes, 194, 455 pyrroloquinoline quinone with hexanol, 188, 281 retinoid isomers, 189, 60, 72 retinoids from interstitial retinol-bindingprotein, 189, 328 retinal pigment epithelial cells, 190, 13 retinol and other nutrients in serum or plasma, 189, 159 and retinyl esters from serum, 189, 98 RNA from mammalian cell nuclei, 180, 78 polyacrylamide gels, 180, 429 subcellular compartments, 180, 12 yeast, 180, 98; 194, 406 rRNA from ribosomes and polysomes, 181, 362

83

FASTP program

simian virus 40 chromosomes, 170, 300 solid-phase, 8-hydroxy-2'-deoxyguanosine in urine, 186, 526 sulfinic acid, 186, 141 thromboxane metabolites for GC/MS assay, 187, 46 tissue lipids, 187, 197 yeast hybrid proteins produced with pATH plasmid, 194, 487 Extravasation leukocytes, intravital microscopic assessment, 186, 733 Extrusion through filters, in preparation of uni- or oligolamellar vesicles, 171,208 Eye, see also specific regions purification of interphotoreceptor retinoid-binding protein, 189,213 vertebrate, interphotoreceptor matrix preparation, 189, 215

F FAB/MS, see Fast atom bombardment mass spectrometry a-Factor degradation, assay, 194, 709 internalization by yeast, assay, 194, 707 radiolabeled, binding to Mata cells, 194, 706 radiolabeling, 194, 702 yeast biosynthesis, 185, 410 leader sequence directed secretion of heterologous proteins, 185, 408 fusions with heterologous proteins, encoding genes, construction,

185,416 hybrid proteins with, expression, 185, 412 secretion signals for directing proteins into secretory pathway, 194, 497 in yeast endocytosis assay, 194, 702 Faraday cage detector in mass spectrometry, 193, 64 FASTA program algorithm, 183, 70 codes for ambiguous nucleotides, 183, 98 file formats, 183, 94 implementation, 183, 67

modifications, 183, 97 scoring matrices alternatives, 183, 92 file, 183, 96 searches for G protein-coupled receptors, 183~ 75 oligonucleotides and oligopeptides, 183, 91 sequence comparisons, 183, 63 Fast atom bombardment mass spectrometry in analysis of posttranslational modification of neuropeptides, 168, 72 analyte modification, 193, 210 artifacts, 193,209 continuous-flow with capillary electrophoresis, 193, 233 carrier solvent composition, 193, 217 ion-suppresion effects, 193, 219 with liquid chromatography, 193, 225 peptides and proteins, 193, 222 probe design, 193, 216 sensitivity, 193, 219 ganglioside GM3derivatives, 179, 249 glycoconjugates, in determination of linkage positions, 193, 587 glycopeptides, 193, 515 glycoproteins, 193, 509 glycosphingolipids, 193, 723 high-resolution, reference compounds~ 193, 875 -HPLC, glycosphingolipids, 193, 725 ionization process, 193, 208 mass reference standards, 193, 311 matrix material spectra, 193, 875 matrix selection, 193, 201 measurements, 2H introduction for, 193, 334 particle beam ionization methods, 193, 26 permethylated carbohydrates, 193, 518 physalaemin-related immunoreactive peptides, 168, 455 sputtering process, 193, 203 -supercritical fluid chromatography, glycosphingolipids, 193, 726 -TLC, glycosphingolipids, 193, 725 FASTP program codes for ambiguous nucleotides, 183, 98 protein sequence comparison, 183, 63

Fatty acids Fatty acids biosynthesis, ~3C NMR monitoring, 177, 429 in endogenous platelet inositol phospholipids, assay, 169, 426 oxo-, nomenclature, 187, 1 protein-bound, acylation, 185, 593 Fc receptors in assay of HL60 cell differentiation, 190, 128 Fenton reaction iron for, sources in vivo, 186, 37 role in hydroxyl radical production, 186, 15 site specificity, 118, 23 Fermentation conditions for design of expression plasmid vectors in Escherichia coli, 185, 34 Ferredoxin component of naphthalene dioxygenase system assay, 188, 49 purification, 188, 50 ~H NMR spectrum, 177, 249 Ferricyanide oxidation of tocopherols, 186, 200 Ferritin -avidin conjugates preparation, 184, 177 in ultrastructural localization of retinal photoreceptor proteins, 184, 370 role in iron metabolism, 186, 33 Ferrocyanide in assessment of glomerular filtration rate, 191, 122 -osmium tetroxide fixation, in visualization of yeast membrane structures, 194, 607 Ferrylbemoglobin absorption spectra, 186, 270 Ferryl species and hydroxyl radicals, distinction, 186, 148 Fibrin autography, in assay of oocyte t-PA, 168, 424 formation, relationship to plateletsurface interactions, 169, 59, 69 and t-PA, recombinant antibody with functions of, production, 178, 515

84 Fibrinogen dark-field electron microscopy, 169, 326 Fibroblasts bovine chemotaxis induced by platelet-derived growth factor assay, 169, 239 checkerboard analysis, 169, 241 counting on chemotaxis membranes, 169, 241 cell cultures, preparation, 190, 176 chicken embryo, vinculin and talin, double fluorescence immunolabeling, 184, 351 CV-1 tightly adherent monolayers, assay of ouabain receptors, 173, 686 whole cells in suspension, ouabain binding, 173, 683 extracts, retinoic acid and retinol metabolism, 189, 525 feeder layer human keratinocyte plating and growth on, 190, 22 preparation for submerged keratinocyte culture, 190, 22 human chemotaxis induced by platelet-derived growth factor assay, 169, 239 checkerboard analysis, 169, 241 counting on chemotaxis membranes, 169, 241 intact, retinoic acid and retinol metabolism, 189, 525 thromboxane Az synthesis, 187, 538 murine adhesion to plastic, 190, 82 attachment to laminin and type IV collagen, enhancement by retinoic acid, 190, 87 bombesin/gastrin-releasing peptide receptors, characterization, 168, 481 cell culture assay for platelet-derived growth factor, 169, 214 cytosolic Ca z+, assay, 168, 490 internalization of bombesin and gastrin-releasing peptide, 168, 489 isolation, 181, 294

85 isolation of 40S hnRNP particles, 181, 294 nuclear extract, preparation, 181, 78 PDGF-dependent growth, effect of gangliosides GM~ and GM~,179, 523 prostaglandin E2 synthesis, 187, 541 overgrowth in primary cultures of islet cells, limitation, 192, 214 whole cell suspensions, amino acid loading of lysosomes, 174, 156 Fibronectin differentiation marker for embryonal carcinoma ceils, 189, 345 Ficoll preparation, 171, 474 Filamentation in Ion mutants, 185, 124 Filaments helical insoluble, composed of hnRNP (A2)~B1 tetramers, generation, 181, 298 Filipin treated renal tissue, freeze-fracture studies, 191, 556 Filter assay carboxypeptidase Y, 194, 659 galactose/N-acetylgalactosamine-binding protein, 179, 323 mannose/L-fucose/N-acetylglucosaminebinding protein, 179, 323 Filters nucleopore, coating, 192, 387 Filtration in assay of fluxes in vesicles, 191, 481 gel, see Gel filtration glomerular assessment, 191, 116 mathematical model, 191, 108 under liquid nitrogen, maize leaf material, 174, 543 membrane, in subcellular fractionation of wheat leaf protoplasts, 174, 526 in NMR sample preparation, 176, 89 ultra-, see Ultrafiltration Fingerprinting double-label, in analysis of protected RNA fragments, 180, 464 peptide, in identification of thrombospondin, 169, 264 RNA

Flip angles ascending RNA homochromatography, 180, 147 first-dimension separation and transfer to thin-layer plates, 180, 137 interpretation, 180, 150 ribonuclease digestion in, 180, 133 role in RNA processing studies, 180, 130 Finger swap assay identification of specific ligands for gene products. 189, 225 Fixation antibodies for immunoelectron microscopy, 194, 619 antibody-nucleosome complexes, 170, 186 platelets, 169, 149 rat brain tissue for receptor immunocytochemistry, 178, 215 Schizosaccharomyces pombe for immunofiuorescence studies, 194, 822 yeast cells for immunoelectron microscopy, 194, 611,614 for immunofluorescence studies, 194, 586, 822 vegetatively grown, procedure. 194, 605 Flank organ hamster, weight, effects of retinoids on sebaceous gland, 190, 328 Flavobacterium meningosepticum purification of mannosyl-glycoprotein endo-/3-Nacetylglucosaminidase, 179, 506 peptide N4-(N-acetyl-/3-glucosaminy[) asparagine amidase, 179, 506 Flavonoids aroxyl radicals, formation and decay, rate constants, 186, 349 radical scavenging efficiencies, 186, 343 Flavoproteins electron-transfer properties, 188, 311 purification from Methylophilus methylotrophus and bacterium W3AI, 188, 309 Flip angles effects in multiple-quantum NMR, 176, 122

Flotation Flotation yeast vacuoles, 194, 655 Flow -force relationships in biological transport, nonequilibrium thermodynamics, 171, 397 laminar, platelet aggregation in, in oitro assay, 169, 138 osmotic, water across red cell membrane, 173, 204 rate in continuous flow tube-based transport assay, constraints, 173, 162 shear, see Shear flow water in plants coupling to other processes, overview, 174, 183 theory, 174, 184 Flow cytometry in assay of DNA, 171, 561 enzyme activity and concentration, 171, 560 proteins, 171, 563 RNA, 171, 561 cell preparation and labeling for, 171, 572 in detection of parathyroid hormone receptors, 184, 279 transforming growth factor-fl receptors, 184, 281 peptide hormone biotinylation for, 184, 275 single ceils, isolation and analysis biological parameters, 171, 559 instrumentation, 171, 551 sample requirements, 171, 564 Fluids, see also specific fluids biological aromatic retinoids and isotretinoin, HPLC, 189, 167 concentration of inhibin from, 168, 616 platelet-activating factor, GC/MS assay, 187, 137 9a,1 lfl-prostaglandin F:, analysis, 187, 54 sulfidopeptide leukotrienes, GC/MS assay, 187, 90 extraceUular analysis of extracellular superoxide dismutase, 186, 260

86 and intracellular, equilibration, 171, 819 human body, lipid peroxide assay, 186, 66 Fluorescamine reactivity in intact cells, in assay of protein degradation, 186, 497 Fluorescein derivatized avidin, preparation, 184, 174 in measurement of pH gradients in cells and vesicles, 191, 472 as pH probe, 174, 132 properties, 174, 132 Fluorescein amine in assay of carbonyl content in oxidatively modified proteins, 186, 476 Fluorescein isothiocyanate labeled dextran in assay of proton pumping in lysosomes and related organelles, 174, 131 as pH indicator, 174, 137 in tracer studies of endocytosis, 191, 559 Fluorescein thiosemicarbazide in assay of carbonyl content in oxidatively modified proteins, 186, 474 N-Fluorescein thioureido-2-mercaptoethylamine derivatization of thiolated Fab, 178, 567 -Fab adduct, in fluorescence quenching binding assay, 178, 567 Fluorescence acridines, in determination of biomembrane surface potential, 171, 367 activated sorting of mammalian cells producing recombinant proteins, 185, 570 amiloride analogs, characteristics, 191, 749 in analysis of nucleosomes, 170, 192 l-anilinonaphthalene 8-sulfonate, in determination of biomembrane surface potential, 171, 370 anisotropy, time-resolved, measurement, 172, 463 auto-, see Autofluorescence in continuous monitoring of transport in cells and vesicles, 172, 122 double, for immunolabeling of antigens, 184, 343

87 ethidium bromide, in assessment of parietal cell permeabilization, 192, 119 excitation ratios, dyes loaded into parietal cells, measurement, 192, 39 Fura-2, spectral properties in situ, 192, 63 4-heptadecyl-7-hydroxycoumarin excitation spectra, 171, 383 titration, 171, 382 in HPLC of arachidonic acid metabolites, 187, 69 immuno-, s e e Immunofluorescence in measurement of plasma membrane potentials, 171, 781 rotational and translational diffusion in membranes, 172, 471 measurement with photomultiplier, TV camera, and digital image processor, 192, 43 microsphere-associated, in detection of cell surface components, 184, 353 in optical potentiometry, calibration to millivolts, 172, 119 phycobiliprotein, in assay of reactive oxygen species and retinoid antioxidant activity, 190, 277 phycoerythrin, in assay of reactive oxygen species, 186, 161 polyadenylate-binding proteins, in determination of binding parameters, 181, 344 poly(1,N6-ethenoadenylic acid), in measurement of dissociation rates, 181, 347 quin2, in assay of cytosolic free Ca 2÷ calibration, 172, 240 recording, 172, 238 signal calibration, 172, 131 sodium-binding benzofuran isophthalate, spectra, 192, 74 substrate concentration range, determination, 172, 131 time-resolved depolarization, in analysis of membrane order and viscosity, 172, 462 2-(p-toluidinyl)naphthalene 6-sulfonate, in determination of biomembrane surface potential, 171, 370

Fluorine electrostatic potential adjacent to membranes, 171, 354 Fluorescence-activated cell sorter in analysis of mammalian cells producing recombinant proteins, 185, 570 in measurement of cellular pH gradients, 172, 93 Fluorescence microscopy in assay of vacuolar pH and cytoplasmic Ca 2- in living cells, 173, 745 hepatocyte couplets, 192, 512 image analysis, 173, 752 instrumentation, 173, 749 measurement principles, 173, 745 nephron segments, 191, 254 pituitary CRF-target cells, 168, 33 yeast cell-associated Lucifer Yellow, 194, 700 yeast cell and nuclear fusion mutants, 194, 777 yeast vacuoles, 194, 645 Fluorescence photobleaching recovery in measurement of translational diffusion in membranes, 172, 496 Fluorescence quenching in binding assay with fluoresceinthiolated Fab adduct, 178, 567 Fluorescence recovery anisotropy in measurement of rotational diffusion in membranes, 172, 488 Fluorescence spectroscopy in analysis of nucleosome structure and dynamics, 170, 198 Fluorescent labeling chicken erythrocyte histone H3 and sea urchin sperm histone H4, 170, 481 nucleosomes, 170, 193 pituitary CRF-target cells, 168, 31 proteins for analysis of protein degradation, 186, 500 Fluorescent staining preelectrophoretic, proteins, 182, 480 Fluorine substituted ligands chemical shift data, interpretation, 177, 16 design, 177, 11 NMR, 177, 3 Tt and T2 data, interpretation, 177, 12 substitution in ligands, consequences, 177, 19

Fluorography Fluorography in one-dimensional electrophoresis of nucleosomes, 170, 121 in two-dimensional electrophoresis of DNA, 170, 132 proteins, 170, 137 Fluorometry N-acetylglucosaminyltransferases III, IV, and V, 179, 397 avidin-biotin interaction, 184, 234 calcium-sensitive indicators in pituitary cells, 168, 275 cytosolic free Ca2+, 172, 230 DNA unwinding, in analysis of DNA strand breaks and repair in mammalian cells, 186, 550 enzymes, substrates, and pathways in isolated nephron segments, 191, 337 hormone effects on free cytosolic Ca-'+ levels, 173, 541 intracellular free Ca2÷, 191, 692 malondialdehyde, 186, 431 membrane potential of murine ascites tumor cells, 172, I00 micro-, s e e Microfluorometry mitochondrial membrane potential in heart during ischemia and reperfusion, 186, 723 Na+/H + exchange, 173, 787 platelet lysosomal acid glycosidase secretion, 169, 340 reactive oxygen species, 186, 161 serotonin secretion by platelets, 169, 208 stilbene disulfonate binding to band 3 protein, 173, 510 tocopherol and tocopheryl ester incorporation and hydrolysis in biomembranes, 186, 355 yeast protease A, 185, 381 Fluorophores in assay of platelet [Ca2+]i, 169, 399 for immunofluorescence studies of yeast, 194, 570 photobleaching, minimization, 194, 576 5-Fluorouridine -anti-idiotypic conjugate, in treatment of murine B cell tumors pharmacological activity, 178, 374 preparation, 178, 372 FMDV, s e e Foot-and-mouth disease virus

88 Follicle-stimulating hormone granulosa cell aromatase bioassay, 168, 414 Follicle-stimulating hormone-releasing protein concentration from biological fluids, 168, 616 iodination, 168, 596 Foods composition tables, for retinol and provitamin A carotenoid content, assessment, 190, 237 malondialdehyde in, HPLC assay, 186, 428 Foot-and-mouth disease virus peptides, with added foreign Th-cell epitopes, immunization with, 178, 666 Footprinting DNase I, in analysis of reconstituted nucleosome core particle authenticity, 170, 580 protein-DNA complexes with y-rays, 186, 545 proteins in analysis of antigen-antibody interactions, 178, 751 in epitope mapping, 178, 759 yeast DNA by primer extension, 194, 550 Formaldehyde developer solution preparation, 194, 728 formation by methanol peroxidation, in assay of drug-dependent HzO2 generation, 186, 622 Formaldehyde dehydrogenase A r t h r o b a c t e r P1 assay, 188, 318 properties, 188, 320 purification, 188, 319 with cofactor requirement assay, 188, 321 properties, 188, 322 purification from R h o d o c o c c u s erythropolis, 188, 322 methylotrophic yeast assay, 188, 455 properties, 188, 457 purification, 188, 456 Formaldehyde dehydrogenase (acceptor),

89 see Aldehyde dehydrogenase (acceptor) Formaldehyde dehydrogenase (glutathione) assays, 188, 315 properties, 188, 317 purification from H a n s e n u l a polymorpha, 188, 316 Formaldehyde transketolase assay, 188, 435 properties, 188, 443 purification from Candida boidinii KD1, 188, 441 Formamide concentration, effect on dsDNA thermal stability, 180, 240 in R N A - D N A hybridization, 180, 363 Formate dehydrogenase Methylosinus trichosporium OB3b assay, 188, 331 properties, 188, 333 purification, 188, 332 methylotrophic yeast assay, 188, 459 properties, 188, 461 purification, 188, 460 Fractionation chromatin, 170, 127 HeLa cells, 181, 321, 328 immuno-, see Immunofractionation immunoglobulin from serum, 182, 669 intercellular, maize leaves, 174, 543 mammalian cell nuclei, 189, 241 mammalian extracts active in pre-mRNA splicing, 181, 3 membrane component-based, in membrane isolation, 172, 13 nonaqueous, lyophilized and ultrasonicated leaf material, 174, 536 potato tuber supernatant, 187, 271 pre-rRNAs, 180, 99 protein bulk precipitates with ammonium sulfate, 182, 293 renal brush border transport proteins, 191, 583 RNA digests on cellulose acetate, 180, 137 size, see Size fractionation sub-, see Subfractionation subcellular Candida organelles containing

Free radicals long-chain alcohol dehydrogenase, 188, 173 long-chain aldehyde dehydrogenase, 188, 177 protoplasts, 174, 520 wheat leaf protoplasts with short quench times, 174, 526 yeast, 182, 171 transport-specific for ATP-dependent Ca 2+ pump purification, 172, 34 integral membrane proteins, 182, 515 yeast DNA by CsCI gradients, 194, 174 yeast nuclei, 194, 746 yeast organelles, 194, 662 Free radical reductase in biological tissues, ESR assay, 186, 670 Free radicals, see also specific radicals absorption spectra, 186, 92 acid-base properties, 186, 92 chain length, measurement, 186, 156 generation by pulse radiolysis, 186, 90 induced damage in Cu2+-ascorbate system, assay, 186, 167 induced products of pyrimidines and purines in DNA, GC/MS analysis. 193, 842 intact cell exposure to, procedure, 189, 489 mediated DNA damage in presence of oxygen, detection, 186, 511 mediated reperfusion injury in renal transplantation, assay, 186, 748 oxygen-derived, role in reperfusion injury, 186, 711 purified protein exposure to, procedure, 186, 489 redox properties, 186, 93 reduction at epidermal surface of skin biopsies, 186, 674 role in human disease, 186, 1 scavenging by flavonoids, efficiency, 186, 343 spin trapping, correlation of radical formation with cell injury, 186, 6 l I sulfur-centered chemical properties, 186, 174 generation, 186, 169 identification, 186, 172

Freeze-~acturing Freeze-fracturing in morphological studies of cellular action of vasopressin, 191, 552 Freeze-thaw processing A m p h i u m a erythrocytes, in measurement of intracellular pH, 173, 333 multilamellar vesicles, 171, 200 Freezing HL60 cells, 190, 122 hybridoma cells, 182, 676 rapid, specimen preparation for electron probe X-ray microanalysis, 172, 205 Frequency analysis k-tuple, intron/exon discrimination and T-cell epitope mapping, 183, 237 Frog, see African clawed frog fl-Fructofuranosidase N-glycosylated, mobility shifts, 194, 685 immunoprecipitation, 194, 694 -prepro human serum albumin fusion cassette, construction, 185,476 yeast mnn mutant, gel electrophoresis, 185, 452 secretion signals for directing proteins into secretory pathway, 194, 497 Fructose-bisphosphate aldolase rabbit muscle, crosslinking, 172, 623 Fructose transporter demonstration in rat adipocytes, 173, 629 Fruit fly cell spreading, solutions for, preparation, 180, 486 chromatin spreading, 180, 487 DNA, purification, 170, 286 embryo chromatin spreading, 180, 490 nuclei and homogenates, preparation, 170, 321 nuclei, isolation, 170, 281 tRNA guanylyltransferase, partial purification, 181, 454 FSH, see Follicle-stimulating hormone Fuchsin -sulfurous acid, for yeast nucleolar staining, 194, 728 Fucoglycosphingolipids structure, biological sources, and properties, 179, 175

90 L-Fucose -mannose/N-acet ylglucosamine-binding protein filter assay, 179, 323 isolation from rat peritoneal macrophages, 179, 324 Fura-2 in analysis of Ca 2÷ metabolism in parietal cells, 192, 67 membrane K ÷ conductance in intestinal secretory cells, 192, 309 in assay of Cai2÷ in parietal cells, 192, 60 platelets, 169, 410 buffering of Ca ~÷, 192, 72 as calcium-sensitive probe, 168, 271 entrapment by mammalian mitochondrial matrix, 174, ll5 loaded cells, C a 2+ signaling, regulating mechanisms, 191, 701 loading of cytoplasm, 173, 763 gastric glands, 192, 63 microfluorimetry, 192, 60 spectral properties in situ, 192, 63 Furosemide related diuretics activity in vitro, evaluation, 191, 790 chemistry, 191, 781 mechanism of action, 191, 784 metabolism, 191, 789 structure-activity relationships, 191, 783

G Galactose -N-acet ylgalactosamine-binding protein filter assay, 179, 323 isolation from rat peritoneal macrophages, 179, 324 ~4C-labeled, in labeling of poly-N-acetyllactosamine glycans, 179, 271 high-copy promoters in yeast, regulated GAL4 expression cassettes for, 185, 297 /3-Galactosidase assay, 189, 267 biotinylation, 184, 154, 474

91 expression in yeast, assays, 185, 356 fusion proteins in protein stabilization, 182, 109 purification, 182, 110 lysosomal marker, assay, 182, 215 secretion from platelet lysosomes, assay, 169, 336 Galactosylceramide 3H-labeled, in assay of glycolipid transfer protein, 179, 559 pyrene-labeled, in assay of glycolipid transfer protein, 179, 561 /3-1,3-Galactosyl-O-glycosyl-glycoprotein /3-1,3-N-acetylglucosaminyltransferase assay, 179, 391 elongation properties, 179, 392 /3-1,3-Galactosyl-O-glycosyl-glycoprotein /3-1,6-N-acetylglucosaminyltransferase assay, 179, 383 properties, 179, 385 Gall bladder Necturus

electrical analog, impedance analysis, 171, 653 epithelium, ion and water transport, electrophysiology, 192, 650 Gangliosides amide-reduced, tandem MS, 193, 764 biological sources, 179, 188 de-N-acetyl-Gm, stimulation of EGF receptor phosphorylation, 179, 531 effect on insulin receptor kinase, 179, 534 Gut anti-ld antibody, in analysis of receptor CT-B ligand-binding domain, 178, 165 effects on PDGF-dependent Swiss 3T3 cell growth, 179, 523 PDGF receptor tyrosine kinase, 179, 523 localization with biotinylated choleragen, 184, 405 G~3 derivatives, synthesis and characterization, 179, 242 effects on EGF-dependent mitogenesis, 179, 525

Gas chromatography EGF receptor phosphorylation, 179. 525 PDGF-dependent Swiss 3T3 cell growth, 179, 523 PDGF receptor tyrosine kinase, 179, 523 protein kinase C of A431 cells, 179. 536 human IgM monoclonal antibodies reacting with, properties and analysis, 179, 294 induction of cell differentiation, overview, 179, 521 HL60 cell differentiation, 179, 538 long-chain bases HPLC, 172, 558 LC/MS, 172, 567 IysO-GM3, inhibition of EGF receptor phosphorylation, 179, 531 native, molecular species, reversedphase HPLC, 172, 552 negative ion tandem MS, 193,750 perbenzoylated derivatives, HPLC, 172, 550 permethylated, tandem MS, 193, 759, 764 properties, 179, 188 regulation of cell proliferation, 179, 521 structure, 179, 188 Gap penalties implementation in Needleman-Wunsch algorithm, 183, 369 position-specific, effects in profile analysis, 183, 157 secondary structure-dependent, in improvement of sequence alignments. 183,408 Gas chromatography in assay for damaged nucleobases, 186, 514 hydrocarbon monooxygenase oxygenation, 188, 4 sugar damage, 186, 519 capillary, retinol, 189, 99 fatty acids in endogenous platelet inositol phospholipids, 169, 426 malondialdehyde, 186, 430 methane monooxygenase, 188, 193 methyl retinoate, 189, 108

Gas chromatography partially methylated alditol acetates, 193, 563 propane oxygenase, 188, 28 propane-specific alcohol dehydrogenase, 188, 23 soluble methane monooxygenase, 188, 181 Gas chromatography-mass spectrometry amino acid-DNA base crosslinks, 193, 850 capillary column preparation, 193, 735 free radical-induced products of pyrimidines and purines in DNA, 193, 842 high-mass, permethylated oligosaccharides, 193, 733 4-hydroxynonenal, 186, 399, 419 and immunoassay, comparison: assay of thromboxane metabolites, 187, 44 isotope dilution, 5-methylcytosine in DNA, 193, 857 lipid hydroxy acids, 186, 683 lipid peroxides, 186, 388 nucleosides, 193, 848 partially methylated anhydroalditols, 193, 576 platelet-activating factor, 187, 134 purine and pyrimidine products in DNA, 193, 846 with selected-ion monitoring in assay of free radical-induced products in DNA and nucleoprotein, 193, 855 oxidative DNA damage, 186, 530 in identification of DNA-protein crosslinks in nucleoproteins, 193, 855 modified bases in DNA, 193, 853 sulfidopeptide leukotrienes in biological fluids, 187, 90 from mast cells, 187, 97 thromboxane metabolites, 187, 42 trimethylsilyl derivatives of nucleic acid constituents, 193, 795 in validation of enzyme immunoassay, 187, 33 Gas chromatography-negative-ion chemical ionization mass spectrometry arachidonic acid metabolites, 187, 69

92 capillary, 6-oxo-prostaglandin F~, 187, 20 epoxy- and dihydroxyeicosatrienoic acids, 187, 179 leukotriene B4, 187, 76 platelet-activating factor, 187, 142 Gas targets for collision-induced dissociation pressure effects, 193, 253 selection, 193, 245 Gastric acid formation in gastric glands, 191, 726 secretion inhibition, relationship to blockade of gastric H+,K+-ATPase, 191,738 related pharmacological agents, analysis, 191, 721 Gastrin effect on gastric glands, 191, 728 Gastrin-releasing peptide -bombesin receptors, in human and murine cells binding studies, 168, 485 characterization, 168, 481 glycoprotein analysis, 168, 487 stimulation of phospatidylinositol turnover, 168, 490 crosslinking studies, 168, 486 detection, 168, 663 internalization by Swiss 3T3 cells, 168, 489 isolation, 168, 676 molecular forms, separation, 168, 665 and related peptides, molecular heterogeneity, 168, 660 radiolabeling, 168, 484 GC/MS, s e e Gas chromatography-mass spectrometry Gel electrophoresis acidic polysaccharides, 179, 95 alcohol dehydrogenase isoenzymes, 189, 438 in analysis of nucleosome spacing periodicity for poly[d(A-T)] chromatin, 170, 600 antibody-induced mobility shifts, in detection of protein in complexes, 181, 279 in assay for propane-specific alcohol dehydrogenase, 188, 23

93 capsular bacterial polysaccharides, 179, 104 carbonyl-fluorescein thiosemicarbazide reaction for, 186, 474 in characterization of nucleosome as chromatin repeat unit, 170, 101 chimeric antibody molecules, 178,473 core histones, 170, 544 crosslinked histones, 170, 554 in demonstration of specific antibodynucleosome interactions, 170, 230 denaturing hybrid polypeptides for antigenic use, 194, 488 yeast hybrid proteins produced with pATH plasmids, 194, 487 in determination of protein purity, 182, 557 DNA fragments, 170, 289, 332 in DNA strand separation, 180, 359 glucose-6-phosphate dehydrogenase, 188, 336 glucose transporter from human erythrocytes, 174, 43 glycoproteins, for blot transfer, 184, 417 heterogeneous nuclear ribonucleoproteins in immunological studies, 181, 324 reconstituted complexes, 181, 313 histone HI, 170, 535 histone subtypes and modified forms, 170, 532 histone subunits characteristics, 170, 473 for preparative purposes, 170,465 for relative molecular weight determination, 170, 469 hydroxypyruvate reductase, 188, 374 in identification of bile acid transport protein in hepatocytes, 174, 28 lg cDNA fragments, 178, 99 in immunoblotting of idiotypes on monoclonal antibodies, 178, 146 in monitoring of nucleosome and chromatin assembly in vitro, 170, 579 native gel modifications for, 180, 449 invertase, in characterization of glycosylation defects, 185, 452

Gel electrophoresis splicing complexes and snRNP particles, 180, 442 nucleoproteins, 170, 634 nucleotide peptides, 170, 407 oligodeoxynucleotide/RNase H digestion products, 181, 291 oligonucleotide primers, 180, 127 one-dimensional gel calibration with marker proteins. 182, 439 nucleosomes, 170, 118 peptides, for mapping, 182, 622 proteins, 182, 425 general procedures, 182, 430 Laemmli method, principles. 182. 429 for peptide mapping, 182. 615 polyadenylated RNAs, 181, 55 polyadenylation-specific complexes, 181. 63 preparative, evaluation, 182. 440 pre-tRNA, 180, 114 protein detection after, 182, 490 protein-DNA complexes, 170. 116 proteins for blot transfer, 184, 417 in determination of size, molecular weight, and presence of subunits. 182, 581 expressed in mammalian cells, 185, 584 for N-terminal sequence analysis. 182, 604, 613 targets in T7 expression system, 185. 85 pulsed field, in positional mapping of yeast genes, 194, 60 retinoic acid-binding protein, cellular form, 189, 299 retinoid-binding proteins, 189, 302 retinol-binding proteins, 189, 299 ribonucleoproteins assembled in vitro. 181, 277 RNA digests, 180, 143 identically sized molecules of different sequence, 170, 356 after modification by diethyl pyrocarbonate or hydrazine, 180, 375 for purification, 180, 377

Gel electrophoresis targets in T7 expression system, 185, 85 small nuclear ribonucleoprotein polypeptides, 181, 259 tRNA splicing endonuclease from Xenopus laevis, 181, 512 two-dimensional crosslinked DNA-protein complexes, 170, 396 crosslinked RNAs, 180, 426 in detection of cAMP-dependent protein phosphorylation, 191, 654 protein degradation in intact cells, 186, 498 specific nucleotide sequences, 170, 129 in determination of histone neighbors in chromatin, 170, 563 DNA, 170, 129 in mapping of protein crosslinking sites on DNA fragment, 170, 401 mini-, proteins, 182, 451 proteins high-resolution methods and applications, 182, 441 for peptide mapping, 182, 616 on Triton-acid-urea gels, 170, 136 RNA, 180, 14 yeast low-molecular-weight RNA, 194, 413 yeast ribosomal proteins in minigel system, 194, 455 tyrosine monooxygenase, 168, 379 viroid and virusoid RNA, 181, 585 von Willebrand factor multimers, 169, 249 yeast DNA, 194, 169 yeast RNA for quality evaluation, 194, 404 Gel filtration with albumin gradient, in isolation of human platelets, 169, 11 in assay of N-acetylglucosaminyltransferase II, 179, 472 sulfobromophthalein binding to bilitranslocase, 174, 51 in concentration of proteins and solute removal, 182, 79

94 in determination of protein purity, 182, 562 glycopeptides, 184, 311 HPLC, glial and neuronal extracts, 168, 559 inhibin, 168, 608 integral membrane proteins, 182, 508 leukocyte arachidonate 5-1ipoxygenase, 187, 315 neurohypophyseal peptides: mono- or diiodinated tracer purification, 168, 579 in NMR sample preparation, 176, 87 plant extracts, 182, 193 in preparation of resealable membranes depleted of cytosolic components, 173, 346 proteins in determination of size, molecular weight, and subunit presence, 182, 579 overview, 182, 317 quinoprotein alcohol dehydrogenase, 188, 35 soluble methane monooxygenase, 188, 185 in washing platelets for use in von Willebrand factor release studies, 169, 247 Gel permeation chromatography in determination of liposome hydrodynamic radius, 171, 215 gastrin-releasing peptides, 168, 668 Gel retardation assay retinoic acid receptor-DNA interactions, 189, 265 Gels, see also specific gels for affinity chromatography of proteins preparation, 182, 361 regeneration, 182, 368 sterilization, 182, 369 storage, 182, 370 for chromatofocusing in different pH ranges, 182, 384 for electrophoresis of nucleosomes preparation, 170, 119 staining, 170, 121 electrophoretic, protein elution, 182, 488 for hydrophobic chromatography of proteins

95 regeneration and storage, 182, 343 selection, 182, 341 for isoelectric focusing casting, 182, 463,468 minigel casting and running, 182,452 preservation, 182, 473 RNA in, recovery, 180, 23 GenBank access, 183, 17 direct submission of data, 183, 11 objectives, 183, 8 restructuring, 183, 15 span and size, 183, 5 status and future directions, 183, 3 Gene mapping in analysis of RNA splicing in Escherichia coil, 181, 526 introns, nuclease digestion method, 180, 325 yeast overview, 194, 38 positional chromosome blotting method, 194, 59 chromosome fragmentation method, 194, 66 Genes amplifiable markers, in heterologous gene selection and coamplification, 185, 542 amplified, variable stability, 185, 539 apr[BamP], based secretion vector, description, 185, 215 avidin, cloning, 184, 71 Bacillus subtilis regulatory, inducible expression, 185, 223 spoOA, placement under inducible control, 185,225 c-fes, 5' end, secondary structure prediction, 183, 292, 296 chimeric antibody, expressing transfectomas, identification and characterization, 178, 469 in chromatin spreads, identification, 180, 508 cloned propagation and expression in yeast with 2-/xm circle-based vectors, 185, 234

Genes templates, RNA polymerase III transcription analysis, 170, 347 translation optimization, 182, 103 T7 RNA polymerase-directed expression in Escherichia coli, 185, 60 coamplification in mammalian cells, 185, 537 for recombinant protein production in mammalian cells, 185, 570 selection protocols, 185, 561 selection vectors for, 185, 560 CUPI, isolation from yeast, 185, 321 dosage, increase for optimization of transcription, 182, 97 eukaryotic branch point sites consensus and conserved sequences, analysis, 183, 264 signal sequences, implications of random distribution, 183, 268 complete, identification problems, 183, 274 evolution, quantitative estimates, effect of base composition, 183. 570 exons identification, 183, 252 and introns, discrimination, k-tuple frequency analysis, 183, 237 potential, prediction, 183, 274 sequence statistics, 183, 252 shuffled, implications for searches of sequence databases, 183, 106 c~-factor leader fusions, construction. 185,416 fusions for expression in Escherichia coli examples, 185, 136 overview, 185, 129 strategies, 185, 133 fusion to strong promoters, 182, 93 GAL4, expression cassette with, construction, function verification, and usage, 185, 297 GAP491, isolation, 185, 342 fl-globin, large intron, effects of point mutations, 181, 45 7r'0-globin, sequence branching: analysis of human evolution, 183, 559 heat-shock protein, induction in yeast, 194, 711

Genes heterologous amplification characteristics, 185, 538 expression in Bacillus subtilis overview, 185, 199 regulated promoter for, 185, 201 expression in mammalian cells experimental approach, 185, 508 overview, 185, 485 vectors for, 185, 487 expression systems, overview, 185, 3 expression in yeast with copper metallothionein promoter, 185, 319 overview, 185, 231 promoters, 185, 425 mRNA analysis, 185, 327 mRNA production in yeast, 185, 422 overexpression, yeast as host for, 182, ll4 -promoter junctions, 185, 427 secretion signal sequences, 185, 427 transcription termination or processing signals, 185, 426 translation in Escherichia coli, enhancing vector, 185, 115 HLS-A3 intron-exon discrimination, 183, 246 sequence information profiles: CpG islands, 183, 242 HO inactive cloned, conversion of homothallic yeast to heterothallism, 194, 144 in mating-type switching, 194, 132 hybrid fusions for heterologous protein secretion by Bacillus subtilis, 185, 217 hydroxybenzoate hydroxylase, related studies, 188, 147 immunoglobulin cloning, 178, 460 rearranged for production of chimeric antibody with antifibrin/t-PA functions cloning, 178, 517 transfection, 178, 523 insulin, transfection of secretory cells, 192, 227 introns and exons, discrimination, k-tuple frequency analysis, 183, 237

96 role in efficient gene expression, i85, 525 in yeast mitochondria, description, 181, 540 keratin expression during vitamin A deficiency of tracheal epithelium, 190, 95 transfection of cultured keratinocytes, 190, 29 lacl, inducible Bacillus subtilis system employing, 185, 211 microprojectile-mediated transfer into mammalian cells, 185, 536 modules, detection, 183, 141 neuroendocrine peptide expression regulation, 168, 618 transcription, assay, 168, 622 neuropeptide, transcription in nervous system tissue, assay, 168, 681 nuclear, for mitochondrial splicing in yeast, screening, 181, 553 overexpression, in studies of Saccharornyces cerevisiae, 194, 239 peptide, expression in diffuse neuroendocrine system, demonstration, 168, 802 PGK, promoter, high-efficiency yeast expression vector based on, 185, 329 pin, phage T4, inhibition of proteolysis in Escherichia coli, 185, 127 polymerase I, II, and III, identification, 180, 497 pre-tRNA, linked to bacteriophage T7 promoter, construction, 180, 64 proinsulin, human, expression with twocistron system, 185, 101 promoter expression, specific mechanisms for control, 185, 48 proteinase inhibitor, induction in plants by di- and triuronides, 179, 569 protein, cloning from purified proteins, 182, 738 rearrangement, genomic divergence through, 183, 428 recombinant viral, detection reagents, 182, 124 reporter, chloramphenicol acetyltransferase, assay, 168, 690 retinoic acid-regulated, expression in

97 murine teratocarcinoma cells, assay, 190, 131 retinol-binding protein, structure and expression, 189,203 rnpA, cells harboring plasmids with, purification of C5 protein, 181, 573 rnpB, cells harboring plasmids with, preparation of M1 RNA, 181, 578 Schizosaccharomyces pombe cloning by gap repair, 194, 815 disruption and replacement, 194, 816 segregation, identification and analysis in yeast with spol3 mutants, 194, 120 spa-BPTl fusion, construction, 185, 155 splice junctions, see Splice junctions Spo-, cloning, 194, 123 sporulation, identification in yeast, 194, ll0 staphylococcal nuclease, placement under control of strong inducible promoter, 177, 76 thymidylate synthase, in analysis of RNA splicing in Escherichia coil, 181, 523 T4 lysozyme, expression system, 177, 46 toxic related plasmid instability, prevention, 185, 80 repression during transcription, 182, 99 trans-acting, for processing yeast cytoplasmic pre-tRNAs, analysis, 181, 400 transfection by electroporation, 178, 468 protoplast fusion, 178, 466 transferred, expression control enhancers, promoters, and splice signals for, assembly, 185, 512 role of introns, 185,525 tRNA processing-related, mutations biochemical consequences, 181,415 genetic analysis, 181, 417 identification, 181,401 suppressor mutations, effects in yeast, 181, 377 transcription conditions, 181, 443 yeast extracts for, 181, 439 trpE fusion

Genes construction with high-expression vectors with multiple cloning sites, 194, 477 design, 185, 162 induction, 185, 164 yeast chromosomal, nomenclature, 194, 8 cloned confirmation, 194, 226 expression, regulation by inorganic phosphate, 194, 385 expression vectors, 194, 373 in vitro mutagenesis and plasmid shuffling, 194, 302 cloning by complementation, 194. 195 disruption methods, 194, 290 episomal wild-type, removal by counterselection, 194, 317 isolation with ,X.gtll system, 194, 230 lethal to Escherichia coil, troubleshooting, 194, 229 manipulation with bacterial transposons, 194, 329 mapped list, 194, 827,835 overexpression in glucose, 194, 387 symbols for, glossary, 194, 830 MAT, HMR, and HML, alleles and mutations, identification, 194, 103 mitochondrial analysis and manipulation, 194, 149 expression, analysis by in vivo labeling of translation products, 194, 156 mutations, isolation, 194, 150 sequence analysis, 194, 160 transformation and replacement, 194, 162 regulating entry into meiosis, identification, 194, 104 replacement methods, 194, 293 RNA polymerase II, transcription in vitro, 194, 545 single, segregation and centromere linkage, 194, 41 sporulation-specific interactions, 194, 109 screening and isolation, 194, 108 Ty mutagenesis, 194, 353 vectors for constitutive and inducible expression, 194. 389

Gentisate 1,2-dioxygenase Gentisate 1,2-dioxygenase assay, 188, 102 properties, 188, 106 purification from Pseudomonas acidovorans, 188, 103 Ginkgolides interactions with platelet-activating factor binding sites, 187, 433 Glass beads column preparation, 169, 108 8-hydroxyquinoline-coated, in generation of metal-free solutions, 186, 124 scanning electron microscopy, 169, 113 in in vitro assay of platelet interaction with artificial surfaces, 169, I08 Glass cuvettes in protein assay, 182, 65 fl-Globin encoding gene, large intron, effects of point mutations, 181, 45 ~r~-Globin encoding gene, sequence branching: analysis of human evolution, 183, 559 Globins multiple sequence alignment and flexible pattern matching, 183, 403 phylogenetic relationships from analysis of 3-D protein structures, 183, 688 simultaneous sequence alignment and phylogeny reconstruction, 183, 639 Glucagon binding to isolated nephron segments, microassay, 191, 315 effects on mitochondria, 174, 123 Glucocorticoids inducible yeast expression vector, 194, 394 induction of cDNA gene expression in mammalian cells, 185,499 2'-O-(fl-o-Glucopyranosyl)-4 ' ,6'-dihydroxy-4-aminodihydrochalcone, see 4Aminophlorizin 2'-O-(fl-o-Glucopyranosyl)-4',6'-dihydroxy-4-azidodihydrochalcone, see 4Azidophlorizin 4'-O-(fl-D-Glucopyranosyl)-2',6'-dihydroxy-4-azidodihydrochalcone, see 4Azido-p-phiorizin 2'-O-(fl-o-Glucopyranosyl)-4',6'-dihy-

98 droxy-4-nitrodihydrochalcone, see 4Nitrophlorizin 2'-(/3-o-Glucopyranosyl-4',6',4-trihydroxy5'-(4-aminobenzyl)-dihydrochalcone, see p-Aminobenzylphlorizin 2'-(fl-D-Glucopyranosyl)-4',6',4-trihydroxy5'-(4-nitrophenylcarbinol)-dihydrochalcone synthesis, 191,760 Glucose overexpression of cloned yeast genes in, 194, 387 in primary culture of islet cells, optimal levels, 192, 216 reabsorption by reptilian proximal tubules, 191, 219 transport in erythrocytes, characterization, 171, 12 Na+-dependent, assay, 192, 445 reconstitution, 192, 443 Glucose oxidase biotinylation, 184, 474 Glucose-6-phosphate dehydrogenase Arthrobacter globiformis

assay, 188, 339 properties, 188, 342 purification, 188, 341 Methylobacillus flagellatum

assay, 188, 335 partial purification, 188, 338 properties, 188, 336 Pseudomonas W6, 188, 347 assay, 188, 346 properties, 188, 348 purification, 188, 347 yeast cytoplasm, assay, 194, 657 Glucose transporter assay and properties, 174, 43 Na+-dependent azide affinity, 191, 767 isolation, 192, 438 partial purification, 191, 596 polypeptide components, partial purification, 191, 603 reconstitution, 192, 438 solubilization, phlorizin binding after, renaturation, 191, 590 purification from human erythrocytes, 174, 39 reconstitution, 174, 39

99 Glucosidase type I assay, 179, 456 properties, 179, 459 purification from mung bean microsomes, 179,457 substrate preparation, 179, 455 type II assay, 179, 46l properties, 179, 463 purification from mung bean microsomes, 179, 462 Glucuronate accumulation by Rhodotorula, energetics, 174, 644 Glucuronides mass spectrometry, 189, 142 in serum extract, HPLC, 189, 140 synthesis, 189, 137 Glutamate accumulation by synaptic vesicles assay, 174, 11 characterization, 174, 16 analogs, synaptic vesicle response, 174, 22 y-carboxylation, 182, 634 carboxyl methylation, 182, 632 Glutamate-ammonia ligase adenylylated, deadenylylation, 182, 799 adenylylation, 182, 795 bicyclic cascade, 182, 801 cascade, identification in Escherichia coli, 182, 793 cumulative feedback inhibition, 182, 794 formation, regulation by UTase and Ptl protein, 182, 807 steady-state mechanisms of regulation, 182, 803 Glutamic acid spin system identification, 177, 139 N~-(~/-Glutamic)lysine crosslink analysis, 182, 636 Glutamine and Lys, differentiation in CID spectra, 193, 468 in proteins, analysis, 182, 600 spin system identification, 177, 139 Glutamine synthetase, see Glutamateammonia ligase Glutaminylpeptide cyclase incubations, 168, 360

Glutathione disulfide radioimmunoassay, 168, 358 substrate preparation, 168, 363 Glutaraldehyde in polymerization of B-cell peptide with T-cell peptide, 178, 664 in preparation of leukotriene C4 immunogens, 187, 118 leukotriene D4 immunogens, 187, 120 Glutathione assay, 186, 757 in bile, assay limitations, 173, 531 in biological samples, assay, 173, 525 cellular, modulation during quinone metabolism, 186, 627 conjugates, transport across hepatocyte plasma membrane, 173, 523 partitioning, 173, 527 defense system against reactive oxygen effects of hypoxia and ischemia. 186, 756 general properties, 186, 752 and glutathione disulfide, partioning of transport, 173, 526 intracellular, assay in mammalian cells. 186, 706 labeled cellular pools, incorporation into leukotriene, 187, 564 plasma, analysis: assay of oxidant stress in vivo, 186, 682 -protein mixed disulfides, quinoneinduced formation, 186, 630 role in quinone-induced depletion of cellular and cytoskeletal protein thiols, 186, 635 and S-conjugate, transport studies, 171, 217 thiyl radical metabolites generation, 186, 318 reactions, 186, 325 transport across hepatocyte plasma membrane, 173, 523 Glutathione disulfide assay, 186, 757 excretion into bile as indicator of oxidative stress, 173, 531 and glutathione, partitioning of transport, 173, 526 plasma, analysis: assay of oxidant stress in vivo, 186, 682 quinone-induced formation, 186, 630

Glutathione disulfide transport across hepatocyte plasma membrane, 173, 523 Glutathione peroxidase role in generation of thiyl radical metabolites of thiols, 186, 323 Glutathione transferase cytosolic and microsomal, and leukotriene C4 synthase, distinction, 187, 311 Glutathionyl groups substituted quinones, half-wave reduction potentials, 186, 191 Glycans anionic, immunobinding, 179, 102 biotinylated derivatives, preparation and assay, 184, 654 structural monitoring, 184, 310 N-Glycans N-acetylglucosaminyltransferases synthesizing, HPLC assay, 179, 351 biosynthesis, 179, 352 structure, 179, 351 O-Glycans N-acetylglucosaminyltransferases synthesizing, HPLC assay, 179, 351 biosynthesis, 179, 357 structure, 179, 352 Glycation effect on Cu-Zn superoxide dismutase in vitro, 179, 575 nonenzymatic, proteins, 182, 645 Glyceraldehyde-3-phosphate dehydrogenase derived expression cassettes for constitutive synthesis of heterologous proteins, 185, 341 plasmid construction, 185, 344 promoter, based expression vector for constitutive gene expression in yeast, 194, 391 Glycerate kinase assay in crude extracts of methylotrophs, 188, 362 Glycerol in buffers for solubilization of membrane proteins, 182, 258 13C-labeled, metabolism: modeling of pentose cycle, 177, 421 disposition on stimulation of phosphatidylinositol breakdown, 191,689

100 formation from diacylglycerol, inhibition by RHC 80267, 191, 690 mass spectra, 193, 876 reference ion masses, 193, 871 Glycerone kinase assay, 188, 451 properties, 188, 453 purification from Candida methylica, 188, 452 Glycine -acetate buffer, nucleotide peptides in, gel electrophoresis, 170, 407 -HCI buffer stock solutions, 182, 31 3H-labeled, in assay of retinoid effects on epidermal renewal, 190, 80 -NaOH buffer stock solutions, 182, 37 proton uptake and K + outflow in presence of, assay, 174, 625 spin system identification, 177, 135 uptake in suspension of washed yeast, assay, 174, 624 Glycine hydroxymethyltransferase assays, 188, 366 detection in serine cycle methylotrophs, 188, 372 glyoxylate-activated, purification, 188, 368 glyoxylate-insensitive, purification, 188, 369 properties, 188, 370 Glycoconjugates associated oligosaccharides isolation, 193, 589 structural characterization, 193, 607 biotinylation on blots, 184, 423 prior to blotting, 184, 420 blotted, direct labeling, 184, 427 fast atom bombardment MS, 193, 591 linkage positions, determination, 193, 587 mass spectrometry, overview, 193, 539 monosaccharide analysis high-performance ion-exchange chromatography with pulsed amperometric detection, 179, 76 sequence-specific, in determination of binding positions, 193, 591 periodate oxidation, 193, 588

101 permethylation and peracetylation, 193, 589 Glycogen synthase, see Starch(bacterial glycogen) synthase Glycoinositol phospholipids membrane anchors of proteins, analysis, 182, 629 Glycolipids affinity chromatography with immobilized lectins, 179, 30 amide reduced, tandem MS analysis, 193,760 fragments in mass spectra, nomenclature, 193,740 mycobacterial antigenicity, 179, 236 isolation, 179, 221 neoantigen synthesis, 179, 239 purification, 179, 225 structural analysis, 179, 229 permethylation, 193, 753 phenolic, purification from Mycobacterium leprae-infected armadillo tissue, 179, 227 reduction by borane, 193, 755 tandem MS analysis, 193, 738 transfer proteins assay, 179, 559 properties, 179, 564 purification from non-porcine sources, 179, 566 porcine brain, 179, 562 Glycopeptides conjugation to proteins, 179, 253 FAB/MS analysis, 193, 515 gel filtration, 184, 311 identification and preparative fractionation, 193, 510 with O-linked saccharides, C1D-MS, 193, 709 triantennary, terminal galactosyl residues, modification, 179, 260 Glycopeptidolipids isolation from Mycobacterium avium, 179, 221 Glycophorin A photochemical labeling, 172, 662 Glycoproteins antibody specificity, 179, 516

Glycoproteins Asn-linked carbohydrates in, attachment site and structural class, 193, 501 biotinylation, 184, 156, 420, 423 on blots, analysis, 184, 415 carbohydrate portion, radiolabeling, 194. 690 cell surface lectin-mediated isolation, 184, 304 O-linked oligosaccharides, characterization, 179, 17 deglycosylation, 182, 537 density gradient purification methods. 182, 535 detection, 182, 535 digestion with glycosidases, 193, 509 FAB/MS analysis, 193, 509 functional role of oligosaccharides. 179. 517 -gold complexes, internalization, 179. 117 histidine-rich antibodies, effect on platelet aggregation, 169, 274 detection in marrow megakaryocytes. 169, 273 ELISA, 169, 269 plasma immunoblot analysis, 169, 273 isolation and purification, 169, 269 platelet detection and assay, 169, 270 secretion, 169, 272 lectin affinity chromatography, 182, 530 myelin-associated, human lgM monoclonal antibodies reacting with. review, 179, 291 neo-, see Neoglycoproteins N-linked chains on. number estimation. 194, 688 nonreducing structures in, mapping. 193. 660 oligosaccharide isolation, 193, 589 platelet, heritable deficiencies, 169, 366 polypeptide backbone, molecular weight determination, 179, 515 reduction and S-carboxymethylation. 193, 508 Saccharomyces cerevisiae

analysis, 194, 682 endoglycosidase H treatment. 194. 694

Glycoproteins [3H]mannose labeling, 194, 695 immunoprecipitation, 194, 694 radiolabeling, 194, 694 selective fragmentation, 193, 508 staining with PAS, 182, 536 thymol-sulfuric acid, 182, 536 substrates in characterization of endoglycosidases, 179, 511 Glycoprotein sialate 7(9)-O-acetyltransferase assay, 179, 417 properties, 179, 420 Glycosaminoglycans isolation, 179, 430 sulfated, ion-exchange chromatography, 179, 433 Glycosidases in analysis of N-linked glycosylation of proteins expressed in mammalian cells, 185, 587 O-linked oligosaccharides, 179, 27 yeast mannoprotein structure, 185, 459 digestion of glycoproteins, 193, 509 Glycoside receptors in cultured heart cells number determination, 173, 643 occupation, correlation with inhibition of active Na+,K + transport, 173, 649 Glycosides acrylylation, in carbohydrate immobilization on polyacrylamide gels, 179, 544 cardiac binding by sodium pump in isolated cells, 173, 678 positive inotropic action, relationship to sodium pump activity in vitro, 173, 655 disaccharidic, derived B2 ions, spectral assignment, 193, 615 monosaccharidic, derived B~ ions, spectral assignment, 193, 610 Glycosphingolipids adsorption to plastic microwells, 179, 546 carbohydrate sequence, 193, 632 ceramide composition, 193, 637

102 derivatization, 193, 626 derivatized reverse-phase HPLC, 172, 556 tandem MS analysis, 193, 759 desorption MS analysis, 193, 713 distillation, 193, 632 intact mixtures, MS analysis, 193, 623 ionization, 193, 722 long-chain bases HPLC, 172, 558 LC/MS, 172, 567 mixture analysis, 193, 726 naturally occurring, core structures, 193, 716 neutral negative ion tandem MS, 193, 748 positive ion tandem MS, 193, 744 structure, biological sources, and properties, 179, 167 nonvertebrate, structure and biological source, 179, 199 oligosaccharide isolation, 193, 590 perbenzoylated derivatives, HPLC, 172, 542 permethylated mixtures, selected-ion curves from MS analysis, 193, 632 permethylated-reduced mixtures, selected-ion curves from MS analysis, 193, 627 permethylation, 193, 589 preparation, 193, 733 separated on TLC plates, cell adhesion, 179, 551 Glycosylation defects in yeast m n n mutants, characterization, 185, 452 heterologous gene products, 185, 436 N-linked, proteins expressed in mammalian cells, 185, 587 O-linked, proteins expressed in mammalian cells, 185, 590 yeast proteins, related properties, analysis, 194, 684 Glycosyltransferases as probes of oligosaccharide structure, 179, 82 Glyoxal removal from RNA, 181, 569 Goblet cells apical membrane

103 isolation from tobacco hornworm midgut, 192,609 purification of vacuolar-type ATPase, 192, 612 Gold -avidin-biotinyl ligand, in visualization of protein trafficking in cell, 184, 384 -avidin, colloid preparation, 184, 381 colloidal adsorption of protein, 179, 114 in agglutination assay of carbohydratespecific binding, 179, 120 in colorimetric assay of proteins, 182, 63 in detection of carbohydrate receptor binding, 179, 111 -glycoprotein complexes, carbohydrate-specific binding and internalization, 179, 117 in immobilization of protein for absorbance, 179, 120 in immunolabeling of yeast ultrathin sections, 194, 619 labeled ligands, dot and Western blotting with, 179, 118 preparation, 179, 112 labeling of histone H2A-H2B dimers, 170, 482 -streptavidin, in ultrastructural localization of retinal pbotoreceptor proteins, 184, 370 Golgi apparatus marker enzyme ~-mannosidase, assay, 182, 219 nucleotide transport, assay, 174, 173 rat liver derived vesicles, preparation and storage, 179,418 membranes, purification of mannosidase IA, 179,446 vesicles, isolation from rat liver, 182, 229 yeast enzyme marker assay, 194, 658 immunofluorescent detection, 194, 601 Gonadotropin-releasing hormone binding to membranal and solubilized receptors, assays, 184, 289 biotinylated

Granulocytes analog description, 184, 396 detection at light microscopic level. 184, 397 localization at electron microscopic level, 184, 401 preparation, 184, 286 dose-response testing by hippocampal brain slice recording, 168, 142 stimulated LH release from pituitary cells, role of protein kinase C, 168, 287 Gonadotropin-releasing hormone receptors binding site, mapping, 168, 729 chemical modification, 168, 730 isolation with avidin-biotin complexes, 184, 287 localization, 184, 395 membranal and solubilized, binding assays, 184, 289 purification, 184, 285 solubilization, 184, 288 Gradient coils in pulse-gradient spin-echo NMR measurement of protein translational motion, 176, 431 Gradient probes in pulse-gradient spin-echo NMR measurement of protein translational motion, 176, 430 Gramicidin channel membranous sites, ion interactions at. NMR studies, 171, 286 NMR-derived and experimental currents, comparison, 171, 336 Granules peptic, isolation, 192, 165 platelet, immunocytochemical localization of proteins, 169, 301 Granules platelet localization of platelet-derived growth factor, 169, 212 proteins, induced chemotaxis of inflammatory cells and fibroblasts. 169, 233 Granulocytes human collection from blood, 186, 228 superoxide radical generation, assay, 186, 586

Granulocytes

104

neutrophilic mature, HL60 cell differentiation into, induction by gangliosides, 179, 538 Granulosa cells rat isolation, 168, 415 metabolic labeling studies in vitro, 179, 439 Growth factors, see also specific growth factors bombesin function as, 168, 492 Growth hormone human export from Escherichia coli, 185, 181 two-site immunochemiluminometric assay, 184, 490 GTP (guanosine 5'-triphosphate labeling to screen RNA for group I introns, 181, 537 Guanidine hydrochloride in isolation of RNA, 190, 134 Guanidine thiocyanate in isolation of RNA, 180, 75 Guanidinium thiocyanate in preparation of RNA, 180, 3 Guanine in RNA, modification by diethyl pyrocarbonate, 150, 374 Guanine nucleotides GTP~/S, caged, synthesis and properties, 172, 289 regulatory proteins, characterization with amiloride, 191, 747 Guanylate enzymatic addition to tRNAHis, 181, 451 Guanylyltransferase in 5' end labeling of RNA, 180, 195 Guard cells isolated, protoplasts and vacuoles patch clamp measurements, 174, 312 preparation, 174, 313

It Halide binding to proteins, quadrupolar NMR study, 177, 330 -H202-peroxidase systems, singlet oxygen generation, assay, 186, 590

Halobacterium halobium purple membrane, solid-state 2H NMR study, 176, 383 Halothane anesthetic for in vivo analysis of opioid peptides, 168, 104 Hamster model for cancer chemoprevention by retinoids, 190, 395 Hansenula polymorpha CBS 4732, purification of alcohol oxidase, 188, 422 methanol metabolizing enzymes, localization, 188, 411 protoplast formation, 188, 413 purification of formaldehyde dehydrogenase (glutathione), 188, 316 Haploidization and recombination in yeast, monitoring, 194, 96 Haptens in obtaining catalytic antibodies conjugation to carrier, 178, 539 design, 178, 532 Haptoglobin monoclonal antibodies in hybridoma cultures, ELISA, 182, 712 Heart chicken embryonic cells, zeugmatin and integrin, double fluorescence immunolabeling, 184, 350 embryonic nonmuscle cells glycoside receptors, analysis, 173, 643 Na+,K÷-ATPase, analysis, 173, 639 preparation and cultivation, 173, 637 ischemia-reoxygenation injury, 186, 74 mammalian, isolated coronary-perfused anaphylaxis, model of eicosanoid release and effects, 187, 619 effects of leukotrienes, 187, 613 platelet-activating factor, 187, 614 prostaglandins, 187, 613 eicosanoid and platelet-activating factor release and effects, 187, 610 ischemia-reperfusion, 187, 616

105 myocardial preservation during transplantation, role of oxygen scavengers, 186, 742 porcine, purification of phospholipid hydroperoxide glutathione peroxidase, 186, 455 rat adult, isolation of Ca2+-tolerant myocytes, 173, 662 isolated working, mitochondrial membrane potential during ischemia and reperfusion, 186, 723 neonatal nonmuscle cells glycoside receptors, number determination, 173, 643 Na+,K+-ATPase, analysis, 173,639 preparation and cultivation, 173, 637 reperfusion injury, role of oxygenderived free radicals, 186, 716 Heat and EDTA, abolition of interference with CAT assay, 168, 695 effect on mammalian cell survival, 186, 700 inactivation of histone-specific RNA processing, 181, 86 induced denaturation, in enzyme purification, 182, 298 Heat shock response in yeast, induction and assay, 194, 710 Heinz bodies formation in intact red blood cells, 186, 272 HeLa cells cell culture for immunopurification of snRNPs, 181, 240 labeling, 180, 413; 181, 321 cell-free transcription extracts efficiencies, 170, 355 preparation, 170, 350 chromatin, preparation, 170, 348 crosslinked ribonucleoproteins in, extraction, 180, 414 isolation of branch points from RNA labeled in vivo with inorganic [32p]phosphate, 180, 178 monolayers

HeLa cells nuclear miniextract preparation, 181. 22 UV irradiation, 180, 413 nuclear extracts active in polyadenylation, preparation, 181, 50 active in pre-mRNA splicing, preparation, 181, 7 characterization, 181, 24 in detection of phosphoproteins, 181, 29 efficient in pre-mRNA splicing, smallscale preparation, 181, 22 fractionation, 181, 69, 87 for histone-specific RNA 3' processing, preparation, 181, 76 for immunopurification of snRNPs. preparation, 181, 239 isolation of snRNP polypeptides, 181. 257 snRNP Sm, 181, 258 preparation, 181, 6 purification of nuclear cap binding proteins, 181, 212 phosphoproteins, 181, 29 with RNA lariat debranching activity, preparation, 181, 183 splicing activities characterization, 181, 16 chromatography, 181, 11 identification, 181.9 nucleoplasm extract preparation and concentration. 181, 266 preparation by cellular fractionation, 181, 321. 328 nucleus isolation for hnRNP particle preparation, 181. 294 pulse labeling of hnRNA, 180, 86 isolation of 40S hnRNP particles, 181. 294 preparation, 1"/0, 348 purification of RNA 3'-terminal phosphate cyclase, 181, 504 radioactively labeled, miniextract preparation, 181, 23

HeLa cells transient transfection for analysis of splicing, 181, 43 Hemacytometry in platelet aggregation assay in vitro, 169, 134 comparison with electronic particle counting, 169, 145 platelet counting, 169, 141 in stirred suspensions, 169, 136, 145 Hemagglutinin reovirus type 3 polyclonal anti-idiotypic antisera in characterization of viral receptor, 178, 325 development and characterization, 178, 322 structural mimicry by anti-idiotypic antibody, 178, 333 Hematoxylin autoxidation in assay of superoxide dismutase, 186, 220 transition metal effects, 186, 223 Heme in assay of soluble cytochromes c from Methylomonas A4, 188, 290 Hemichrome absorption spectra, 186, 270 Hemitoxins modified anti-idiotypic antibodies, in treatment of B-cell neoplasms, 178, 356 Hemocyanin keyhole limpet, see Keyhole limpet hemocyanin Hemoglobin absorption spectroscopy, 186, 268 amino acid sequences evolutionary tree construction, 183, 604 progressive alignment and phylogenetic tree construction, 183, 380 Heinz body formation in intact red blood cells, 186, 272 oriented crystals, solid-state 2H NMR, 176, 382 oxidative reactions, 186, 265 preparation, 186, 266 reactions with xenobiotics, 186, 267 role in iron metabolism, 186, 34

106 thiol oxidation, 186, 271 translational motion, measurement by pulse-gradient spin-echo NMR, 176, 439 Hemolymph Allomyrina dichotoma, purification of lectins, 179, 332 Hemolysin based system for secretion of heterologons proteins from Escherichia coli, 182, 132 secretion pathway in Escherichia coli, 185, 173 Hemorrhage induced noradrenaline release in supraoptic nucleus, microdialysis assay, 168, 201 Hemosiderin role in iron metabolism, 186, 33 Heparin treatment of nuclear extracts for native gel electrophoresis, 180, 447 Hepatitis B virus core protein, presentation of peptide epitopes on, 178, 669 protein antibodies, synthetic T-cell epitopes as immunogens for, 178, 634 Hepatitis 8 virus satellite RNAs, self-cleaving sequences, 180, 556 Hepatocytes, see also Liver Ca2÷ fluxes and phosphoinositides, assay, 173, 534 couplets electrophysiology, 192, 514 fluorescence microscopy, 192, 512 isolated, functional characteristics, 192, 505 isolation, 192, 501 microinjection methods, 192, 511 cytosolic free Ca2÷ levels, hormone effects, 173, 541 glutathione and protein thiol status during quinone metabolism, 186, 627 isolated, alanine transport, 174, 32 isolation, 173, 567 loading with myo[2-3H]inositol, 173, 535 monolayer cultures

107

Hexose

amino acid transport, assay, 173, 564 preparation, 173, 567 plasma membrane alanine-transportingproteins, identification, 174, 38 sinusoidal, identification of bile acid transport protein, 174, 25 transport of alanine, 174, 31 glutathione, glutathione disulfide, and glutathione conjugates, 173, 523 purification from liver cell isolates, 190, 55 rat isolation, 173, 535; 187, 278 leukotriene metabolism, 187, 277 plasma membrane biogenesis, 191, 825 purification, 187, 278 suspension cultures amino acid transport, assay, 173, 564 preparation, 173, 567 4-Heptadecyl-7-hydroxycoumarin excitation spectra, 171, 383 fluorescence titration, 171, 382 Herpes simplex virus detection and typing with avidin-biotin reagents, 184, 541 Heterologous proteins constitutive synthesis, GAPDH expression cassettes for, 185, 341 expression effects of promoter length and orientation, 185, 345 in Escherichia coli, 185, 11 in Saccharomyces cerevisiae with ADH2 promoter, 185, 308 a-factor fusions with, encoding genes, construction, 185, 416 glycosylation, 185,436 modification by yeast, 185, 232 N-terminal analysis by radiolabeling, 185, 221 precursor synthesis in Bacillus subtilis, demonstration, 185,220 secretion by Bacillus subtilis, 185,214 Escherichia coli

genetic engineering for, 185, 166

hemolysin secretory pathway, 182. 132 yeast enhancement by molecular and genetic methods, 185,471 a-factor leader-directed, 185,408 with heterologous and homologous signal sequences, 185, 421 overview, 185,232 plasmids for, components, 185,422 regulated, yeast expression plasmid allowing, construction, 185, 312 signal peptide-processing site, determination, 185, 220 stability after secretion by Bacillus subtilis, 185, 200 in yeast, 185,233 Heuristic refinement in determination of protein solution structure from NMR data, 177, 218 Hexadecanol dehydrogenase NAD+-dependent, membrane-bound assay, 188, 17 properties, 188, 18 1-O-Hexadecyl-2-arachidonoyl-glycerophosphocholine 3H-labeled, preparation, 187, 217 Hexadimethrine bromide in DNA transfection of mammalian cells, 185, 532 Hexanol extraction of pyrroloquinoline quinone, 188, 281 Hexapole reaction cells in tandem mass spectrometry, 193, 169 Hexosamines radiolabeled, specific activity in metabolic labeling studies, 179, 435 fl-Hexosaminidase, see fl-N-Acetylhexosaminidase Hexose transport in suspended cells assay, 173, 616 as control point, evaluation, 173,633 modulation, 173, 631 substrate concentration dependence, 173, 624 uptake by Chlorella, inducibility and specificity, 174, 391

Hexose monophosphate shunt Hexose monophosphate shunt based assay for HL60 cell differentiation, 190, 127 3-Hexulose-6-phosphate synthase Acetobacter methanolicus MB58 assay, 188, 401 properties, 188, 403 purification, 188, 402 Bacillus CI assay, 188, 392 properties, 188, 395 purification, 188, 393 Mycobacterium gastri MBI9 assay, 188, 397 properties, 188, 400 purification, 188, 399 High-performance gel chromatography in determination of membrane protein molecular weight, 172, 514 High-performance liquid chromatography N-acetylglucosaminyltransferases involved in N- and O-glycan synthesis, 179, 351 types III, IV, and V, 179, 397 angiotensin II, 168, 550 and antibodies, in analysis of prohormone processing, 168, 517 arachidonate-containingphosphoglycerides, 187, 160, 164 arachidonate 5-1ipoxygenase leukocyte, 187, 312 potato, 187, 299 aromatic retinoids in biological fluids, 189, 167 bonded-phase, prostaglandin H2, 187, 249 caged nucleotides, 172, 295 a- and B-carotenes and other nutrients in serum or plasma, simultaneous analysis, 189, 155 catecholamines, 168, 439 catechol 1,2-dioxygenase, 188, 124 -chemical ionization MS, glycosphingolipids, 193, 725 in chiral analysis of eicosanoids, 187, 187 chlorocatechol 1,2-dioxygenase, 188, 125 cholesterol 5,6-epoxides, 186, 441 cytochrome P450 arachidonate oxygenase, 187, 388

108 2'-deoxycytidine photoproducts, 186, 508 5(S),12(S)-dihydroxy-6,10-trans-8,14-ciseicosatetraenoic acid, 187, 305 12S-20-dihydroxyeicosatetraenoic acid production by neutrophils, 187, 594 N-dinitrnphenyl-S-carboxymethylaminothiols, 186, 707 with electrochemical detection in analysis of substituent effects on quinone half-wave reduction potentials, 186, 187 8-hydroxy-2'-deoxyguanosine in DNA and urine, 186, 521 quinones, 186, 180 -FAB/MS, glycosphingolipids, 193, 725 with fluorescence detection, arachidonic acid metabolites, 187, 69 gangliosides, 172, 550 glucuronides in serum extract, 189, 140 glycosphingolipids, 172, 542 hepatic retinyl ester hydrolase, 189, 492 homoprotocatechuate enriched with [170] or [1sO]hydroxyls, 188, 113 hydrophobic interaction, arachidonate 5lipoxygenase, 187, 271 12S-hydroxyeicosatetraenoic- 1,20-dioic acid production by neutrophils, 187, 597 4-hydroxynonenal, 186, 419 inositol phosphates, 173, 537 intracellular glutathione, 186, 706 isocratic adsorption, retinyl esters and geometric isomers, 189, 181 isocratic, bile pigments, 186, 309 isocratic ion-exchange, inositol phosphate isomers, 191, 711 with isoluminal chemiluminescence detection, lipid hydroperoxides, 186, 371 isotretinoin in biological fluids, 189, 167 lecithin-retinol acyltransferase, 189, 454 leukocyte arachidonate 5-1ipoxygenase, 187, 315 leukotriene A4 methyl ester, 187, 287 linoleic acid hydroperoxide, 186, 308 lipid hydroperoxides, 186, 308 lipoxins, 187, 167 lipoxygenase products of arachidonic acid, 187, 98

109

H i g h - p e r f o r m a n c e liquid c h r o m a t o g r a p h y

leukotrienes, 187, 570 long-chain bases of membrane lipids, 172, 558 lycopene and other nutrients in serum or plasma, simultaneous analysis, 189, 155 malondialdehyde, 186, 413,428 -mass spectrometry, mass reference standards, 193, 310 membrane lipids, 172, 538 methyl linoleate hydroperoxides and dienols, 186, 381 neuroendocrine peptides extracted from milk, 168, 496 nuclear extracts, 189, 245 nuclear retinoic acid receptors, 189, 249 peptides for mapping, 182, 625 N-terminal sequence analysis, 182, 607 phosphatidylcholine hydroperoxides, 186, 159, 308 phospholipids, 172, 560 proteins chromatographic modes, 182, 399 on immobilized reactive dyes, 182, 355 instrumentation, 182, 394 for N-terminal sequence analysis, 182, 607 retinal oxime isomers, 189, 73 retinal produced in hepatic microsomes, 189, 520 retinoic acid analogs, 189, 18, 20 retinoid isomers, 189, 60, 70 retinoids applied locally to embryos, 190, 207 with automated column switching, 189, 146 in blood, 189, 50 extracted from interstitial retinolbinding protein, 189, 328 for metabolic studies, 189, 529 natural and synthetic, in human skin samples, 190, 163 quantitative analysis, 190, 9 synthesized from retinol and/3-carotene, 189, 474 retinol isomers, 189, 73 retinol metabolites produced by microsomes, 189, 485 retinol and other nutrients in serum or

plasma, simultaneous analysis, 189, 155 retinyl ester isomers, 189, 74 retinyl esters, 189, 81 reversed-phase angiotensin metabolites, 168, 563 arachidonic acid metabolites, 187. 370. 378 column selection, 193, 399 cytochrome P450 prostaglandin cohydroxylase, 187, 260 derivatized glycosphingolipids, 172, 556 enzymatic digest and protein chemical cleavage products, 193, 389 in estimation of peptide phosphorylated/dephosphorylated ratio, 193. 490 gastrin-releasing peptides, 168, 671 glycopeptides, 193, 510 gradient times and flow rates. 193, 402 inhibin, 168, 609 leukotriene B4, 187, 557 in leukotriene B4 inactivation assay. 187, 322 leukotriene C4 synthase, 187, 307 leukotriene metabolites, 187, 281 long-chain bases of membrane lipids, 172, 559 material cross-reacting with angiotensin antibodies, 168, 559 mobile phase considerations, 193, 398 monosialogangliosides, 172, 552 native gangliosides, 172, 552 parathyroid hormone, 184, 277 peptides, 193, 407 performance criteria, 193, 391 phospholipid molecular species, 172, 563 in protein concentration and solute removal, 182, 80 proteins, 182, 414 recombinant proteins, 193, 522 retinyl esters, 189, 85 sample volume and buffer selection. 193, 404 sphingomyelin, 172, 563 system selection, 193, 392 trypsin-cleaved prostaglandin endoperoxide synthase, 187, 483

High-performance liquid chromatography tryptic digests of reductively methylated histones, 170, 381 RNA hydrolysates, retention times, 193, 803 size exclusion in analysis of protein and peptide epitopes antibody-peptide interactions, 178, 122 in binary complexes, 178, 108 in ternary complexes, 178, 116 LH-RH precursor, 168, 530, 532 proteins, 182, 399 retinoid-bindingproteins, 190, 445 spin-labeled proteins, 177, 105 thymidine photoproducts, 186, 504 tocopherols and other nutrients in serum or plasma, simultaneous analysis, 189, 155 vasopressin metabolites, 168, 388, 394 xanthine oxidase, 168, 654 High Powder Azure in overlay test for yeast protease B activity, 194, 435 Hippocampus slices, in dose-response testing of peptides, 168, 129 H i r u d o m e d i c i n a l i s , s e e Leech Histamine effects on gastric glands, 191, 728 prostaglandin Ez production by parietal cells, 187, 513 iodination and coupling to biotin, 184, 213 Histidine -copper complexes, preparation, 168, 711 iodination and coupling to biotin, 184, 217 spin labeling in proteins, 177, 116 spin system identification, 177, 138 tRNA, enzymatic addition of guanylate, 181, 451 Histidylproline diketopiperazine radioimmunoassay, 168, 572 Histochemistry hybridization, s e e Hybridization histochemistry

1I0

immuno-, s e e Immunohistochemistry lectins, 184, 361 Histones antibodies, preparation for nucleosome studies, 170, 251 blotting onto glass fiber membranes, 170, 475 in chromatin neighbors, determination, 170, 563 oligomeric state, determination, 170, 557 core electrophoresis, 170, 544 H1 histone separation from, 170, 526 preparation, 170, 587 crosslinked, gel electrophoresis, 170, 554 crosslinking, 170, 549 derivatives, preparation, 170, 480 HI amino acid sequences, 170, 504 electrophoresis, 170, 535 nucleosomes with, localization of 5methylcytosine, 170, 248 preparation, 170, 590 purification and analysis, 170, 524 structural role in nucleosomes, 170, 104 variants, regular set linker histone separation from, 170, 528 resolution, 170, 529 H2A amino acid sequences, 170, 508 -H2B dimers, gold labeling, 170, 482 H2B amino acid sequences, 170, 512 -H2A dimers, gold labeling, 170, 482 H3 amino acid sequences, 170, 516 chicken erythrocyte, fluorescent labeling, 170, 481 H4 amino acid sequences, 170, 520 sea urchin sperm, fluorescent labeling, 170, 481 H5 amino acid sequences, 170, 504 -H5 crosslinks, assignment to specific H5 domains, 170, 565

111

Homonuclear Hartmann-Hahn spectroscopy

preparation, 170, 590 strong nucleosome binding site, radiolabeling, 170, 376 immunogens for antihistone antibodies, preparation, 170, 251 isolation and characterization, 170, 431 linker, separation from regular set of H1 variants, 170, 528 mRNA, 3' processing in mammalian cell nuclear extracts, 181, 74 natural complexes, isolation, 170, 441 octamers hybrid, reconstitution, 170, 484 natural core particle assembly from, 170. 493 reconstitution, 170, 484 in nucleosome core, Lys-containing DNA-binding regions, radiolabeling, 170, 374 reconstituted core particle assembly from, 170, 493 crystallization, 170, 491 structural confirmation, 170, 486 pairs, purification from chromatin, 182, 335 peptides, as immunogens for antihistone antibodies, preparation, 170, 253 preparation from chicken erythrocytes and murine liver, 170, 632 HeLa or HTC cells, 170, 623 isolated nuclei, 170, 534 reductively methylated tryptic digestion and peptide mapping, 170, 379 tryptic peptides fractionation and purification, 170, 381 radiolabeled, identification and specific radioactivities, 170, 383 relative affinity for specific DNA fragments, measurement, 170, 642 in solution, oligomeric state, determination, 170, 557 subtypes and modified forms, gel electrophoresis, 170, 532 subunits

chemical characterization, 170, 469 purification, 170, 449 relative molecular weights, determination, 170, 469 transfer to DNA fragments, 170, 578 HIV, s e e Human immunodeficiency virus HOHAHA, s e e Homonuclear HartmannHahn spectroscopy Holotoxins modified anti-idiotypic antibodies, in treatment of B-cell neoplasms, 178. 356 Holzer's "inactivase" identification as adenylyltransferase. 182, 798 Homeostasis associated compensatory mechanisms, maintenance in e x vivo studies, 169. 98 Homochromatography ascending, in RNA fingerprinting, 180. 147 Homogenization in cell disruption, 172, 5 control, 172, 8 liver, 174, 54 media for, 172, 7 partial, maize leaves, 174, 543 in production of small unilamellar vesicles, 171, 203 starting material selection, 172, 5 tissue, in subcellular membrane isolation, 172, 19 Homonuclear correlated spectroscopy high-order experiments, 176, 132 two-dimensional metabolite detection in vivo, 176, 336. 493 multiple-quantum-filtered, proteins. 176, 118 two- and three-quantum-filtered experiments, 176, 124 Homonuclear Hartmann-Hahn spectroscopy hardware, 176, 167 interference from NOE, 176, 159 mixing based on composite pulse decoupiing schemes, 176, 156 peptides and proteins, 176, 159

Homonuclear Hartmann-Hahn spectroscopy principles, 176, 152 relaxation during mixing, reduction, 176, 158 spectra recording without presaturation, 176, 164 tracing connectivities along aliphatic side chains, 176, 162 in 2D NMR of proteins, 176, 33 Homonuclear Hartmann-Hahn transfer suppression in rotating-frame NOE, 176, 213 Homoprotocatechuate [170] or [ISO]carboxyl-enriched, synthesis, 188, 110 [170] or [I80]hydroxyl-enriched, enzymatic synthesis, 188, 113 Hormone receptors, see also specific h o r m o n e receptors

affinity purification, 184, 636 anti-idiotype-binding, structure, 178, 207 detection and analysis in isolated renal tubules, 191, 303 -hormone coupling, analysis, 172, 605 isolation and characterization, 184, 244 structure and function, anti-idiotypic antibodies as probes, 178, 191 Hormones, see also specific h o r m o n e s activation of cAMP-dependent protein kinase, detection in gastric parietal cells, 191, 652 a-adrenergic agonists, effects on mitochondria, 174, 119 antidiuretic, see Vasopressin binding to isolated nephron segments, microassay, 191, 315 biosynthesis by cultured islet cells, 192, 219 control of sinusoidal glutathione transport, 173, 528 diuretic, bioassay, 191, 178 effects on free cytosolic Ca 2÷ levels in hepatocytes, 173, 541 intramitochondrial Ca 2+, 174, 111 exocrine activation, patch-clamp studies, 171, 663 peptide, biotinylation, 184, 275 pituitary, release from single cells, assay, 168, 266 -receptor coupling, analysis, 172, 605

112

release by cultured and intact islet cells, 192, 217 secretion basal and pulsatile, regulation, analysis by passive immunoneutralization, 168, 498 from single cells, cell blot assay, 168, 327 Horseradish peroxidase -avidin-biotin, in immunoassay of HIV core proteins, 184, 556 -avidin, in detection of biotinylated proteins, 184, 435 leukocyte surface proteins, 184, 431 biotinylation, 184, 150 endosomal marker assay, 182, 217 labeling of endosomal compartment, 182, 216 -streptavidin, in detection of antigens on Western blots, 184, 441 biotinylated proteins, 184, 435 in tracer studies of endocytosis, 191, 557 Host specificity for ascitic cells, 171, 600 HPLC, see High-performance liquid chromatography 5-HT, see Serotonin Human immunodeficiency virus core proteins, immunoassay, 184, 556 derived synthetic peptide antigens, 178, 708 gpl60 peptides, overlapping studies of immunogens, 178, 627 T-cell antigenic sites, identification, 178, 622 type 1, active reverse transcriptase, expression in Bacillus subtilis, 185, 210 types 1 and 2, infections, distinguishing immunoassays, 178,693 Hyaluronic acid preparation, 179, 283 Hybridization colony, in screening for M A T a 2 oligonucleotide-containing clones, 185, 361 DNA fragments, 170, 335 dot-blot, after direct incorporation of biotin into DNA, 184, 610

113 liquid, in determination of nucleasesensitive regions of chromatin, 170, 340 in mapping of yeast genes, 194, 47 in neuroendocrine peptide hnRNA and mRNA assay, 168, 626 in neuropeptide gene transcription assay in nervous system tissue, 168, 685 Northern in analysis of tRNA mutants in vivo, 181, 388 in electrophoretic detection of ribonucleoproteins, 180, 451 Northern and dot blots, 168, 407 -nuclease protection assay, polyadenylation-specific events, 181, 57 nucleic acids probe labeling, 184, 577 in 2-D electrophoresis of DNA, 170, 132 oligonucleotide mixtures in sodium chloride-sodium citrate, 182, 750 tetramethylammonium chloride, 182, 748 photobiotin application, conditions, 184, 595 in positional mapping of yeast genes, 194, 63 protein image, in identification of crosslinked proteins and DNA, 170, 396 RNA-DNA with dsDNA probes, 180, 343 for primer extension analysis of RNA aqueous methods, 180, 363 hybridization with formamide, 180, 363 primer selection and preparation, 180, 352 theory and controls, 180, 361 principles and reaction conditions, 180, 242 in pulse labeling of hnRNA, 180, 92 by R-loop formation, 180, 250 by RNA-ssDNA heteroduplex formation, 180, 251 with ssDNA probes, 180, 340 in screening for arachidonate 5-1ipoxygenase clones, 187, 495

Hybridization histochemistry in situ

in analysis of active genes and transcript configuration, 180, 508 in assay of vasopressin and oxytocin mRNA in brain, 168, 409 autoradiographic signals, image analysis, 168, 410 data analysis, graphical and statistical methods, 168, 822 demonstration of peptide gene expression in diffuse neuroendocrine system, 168, 802 in localization of cellular retinol- and retinoic acid-binding protein mRNA, 189, 321 nonradioactive, in visualization of neuroendocrine mRNAs, 168, 753 quantitation with radiolabeled probes, 168, 741 and quantitative autoradiography, data analysis, 168, 825 retinoid-binding protein mRNA, 189, 286 3-~S-labeled probes ssDNA, 168, 402 ssRNA, 168, 403 solution in assay of oxytocin and vasopressin mRNA in brain, 168, 404 35S-labeled ssDNA as probes, 168, 402 -nuclease protection assay differentially spliced substance P and neurokinin A mRNAs, 168, 634 uniformly labeled cRNAs in, 168, 645 Hybridization histochemistry in situ

combination with markers of neuronal connectivity, 168,778 detection of neuropeptide mRNA with DNA and RNA probes, 168,702 high-resolution methods, 168, 761 RNA probes, 168, 771 synthetic oligonucleotides probes, 168, 763 probes and probe labels, selection, 168, 785 quantitation and digital representation, 168, 808

Hybridomas Hybridomas for catalytic antibody production, preparation and screening, 178, 541 cell freezing, 182, 676 cultures for production of 12CA5 and 9El0 antibodies, 194, 511 culture supernatant, biotinylation of monoclonal antibodies in, 184, 472 monoclonal haptoglobin antibodies, ELISA, 182, 712 production by facilitated cell fusion, 184, 641 screening for renal cell-specific monoclonal antibodies, 171, 583 by solid-phase enzyme immunoassay, 178, 54 by solid-phase free ligand immunoassay, 178, 52 secreting chimeric antifibrin antibody with t-PA activity, selection, 178, 523 subcloning, 182, 676 HYBRIP.BA program analysis of transport kinetics, 172, 270 Hydration after dehydration multilamellar vesicles, 171, 199 in orientation of synthetic and native membranes, 172, 403 lipids, 171, 195, 198 Hydraulic conductivity absolute value and physiological meaning, 174, 193 determination with pressure probe, 174, 354 from single time course, 173, 220 for osmotic water flow across red cell membrane, determination, 173, 208 pressure dependence, 174, 197 proximal tubules from reptile, estimation, 191, 206 Hydrazine in assay of covalently bound pyrroloquinoline quinone, 188, 277 in modification interference analysis of reactions with RNA substrates, 180, 369 Hydrazinolysis mannoproteins, 185, 467

114 Hydrocarbon monooxygenase cell-free production of oxygenated products, 188, 7 isolation from P s e u d o m o n a s oleooorans, 188, 7 oxygenation activity, assay, 188, 4 reductase component, isolation, 188, 6 rubredoxin component isolation, 188, 5 properties, 188, 8 Hydrocarbons polycyclic aromatic, see Polycyclic aromatic hydrocarbons Hydrochloric acid and acetone, in precipitation of proteins, 182, 592 amino sugar hydrolysis in, 179, 81 Hydrodynamic radii liposomes, determination with gel permeation chromatography, 171, 215 Hydrogen, see also Protons and carbohydrates, cotransport in Rhodotorula, 174, 629 deuteron-proton exchange, in NMR sample preparation, 176, 81 exchange labeling, in characterization of folding pathways, 176, 465 exchange, and protein stability correlation, 176, 454 kinetic limits, 176, 452 structural unfolding model, 176, 451 ~H, spin-lattice relaxation, 176, 188 -hydrogen correlations in 2D NMR of proteins, 176, 28 Hydrogen chloride -glycine buffer stock solutions, 182, 31 Hydrogen peroxide decomposition, inhibition, 186, 149 drug-dependent generation, assay in vitro, 186, 621 -peroxidase-halide systems, assay of IO2 generation, 186, 590 reactions with hemoglobin, 186, 267 metal-polyaminocarboxylatecomplexes, 186, 117 role in human disease, 186, 8 Hydrolysis amino sugars in hydrochloric acid, 179, 81

115 arachidonic acid, tritiated, in assay of phospholipase A2, 187, 216 ATP, dependence of light-induced hyperpolarization, 174, 487 2,2-dimethyloxirane by epoxide hydrolase, mechanistic studies, 177, 387 DNA for analysis of 5-methylcytosine, 193, 86O for GC/MS, 186, 531; 193, 843 radical-damaged, 186, 512 enzymatic DNA and RNA for MS analysis, 193, 782 peptide esters, 193, 347 esterified chromanols in membrane suspensions, 186, 358 incomplete, membrane-permeant dyes, 192, 46 inositol lipids, 168, 343 leukotriene A4 hepatic enzymes catalyzing, purification and properties, 187, 324 product characterization, 187, 325 leukotriene A4 methyl ester, 187, 308 lysosomal, proteins in proximal tubules, 191, 340 methylated carbohydrates for a!ditol acetate preparation, 193, 559 neutral sugars in trifluoroacetic acid, 179, 80 nucleoproteins, 186, 531; 193, 843 phosphoinositides in isolated cell membranes, assay, 168, 338 phospholipase C-catalyzed, phosphoinositides in suspension, assay, 187, 230 phospholipids by phospholipase C, 187, 206 proteins, for amino acid analysis, 182, 594 Hydropathy functions correlation, 172, 444 Hydropathy plots in analysis of hydrophobic membrane protein folding patterns, 172, 440 cytochrome f, effect of averaging window, 172, 448 Hydropathy scales comparison, 183, 454

HYDRO program Hydroperoxidation enzymatic, soybean phosphatidylcholine, 186, 453 Hydroperoxides assay with diphenyl-l-pyrenylphosphine. 186, 157 cyclooxygenase initiation assay, 186. 431 in DNA, functional determination, 186. 519 iodometric assay, 186, 434 reductive cleavage by cytochrome P450. 186, 278 synthesis, 186, 433 Hydroperoxy acids nomenclature, 187, 1 5(S )-Hydroperoxyeicosatetraenoic acid preparation, 187, 302 Hydroperoxyl radicals role in human disease, 186, 10 Hydrophile-lipophile balance number detergents, 182, 244 Hydrophilicity plotting in identification of protein antigenic segments and other interaction sites, 178, 571 Hydrophobic interaction chromatography ceramide glycanase, 179, 481 high-performance liquid, proteins, 182, 409 proteins, 182, 339 column running, 182, 342 gel regeneration and storage, 182, 343 gel selection, 182, 341 protocatechuate 3,4-dioxygenase, 188, 86 and reversed-phase chromatography, comparison, 182, 340 RNA, 180, 33 Hydrophobicity effects in crosslinking studies, 172, 585 in isolation of yeast spores, 194, 147 proteins, effects of oxidative modification, 186, 493 Hydrophobic moment in colicins, 172, 457 HYDRO program hydrophilicity analysis of protein antigenic segments and other interaction sites, 178, 578

Hydroxide Hydroxide -Fe(IV) complex, generation in aqueous solutions, 186, 109 Hydroxy acids nomenclature, 187, 1 4-Hydroxyalkenals assay, 186, 405 binding to proteins, 186, 405 extraction from tissue, 186, 402 synthetic, analysis, 186, 403 Hydroxyapatite, see Durapatite Hydroxybenzoate hydroxylase assay, 188, 139 genetics, 188, 147 properties, 188, 142 purification from P s e u d o m o n a s aeruginosa, 188, 139 8-Hydroxy-2'-deoxyguanosine in DNA and urine, HPLC with electrochemical detection, 186, 521 12(R)-Hydroxy-5,8,10,14-eicosatetraenoic acid bioassay, 187, 378 12S-Hydroxyeicosatetraenoic- 1,20-dioic acid production by neutrophils, assay, 187, 597 12(R)-Hydroxy-5,8,14-eicosatrienoic acid bioassay, 187, 378 2-Hydroxyestradiol role as antioxidant, 186, 330 Hydroxylamine induction of yeast mutagenesis in vitro, 194, 315 Hydroxylapatite, see Durapatite o~-Hydroxylase component of Pseudornonas oleovorans hydrocarbon monooxygenase, cellfree production of products, 188, 7 Hydroxylation salicylate by dye-mediated hydroxyl radical generation, 186, 638 toluene by cumyl hydroperoxide, assay, 186, 275 Hydroxyl groups substituted quinones, half-wave reduction potentials, 186, 190, 192 Hydroxyl radicals deoxyribose assay, 186, 26

116 detection and assay with dimethyl sulfoxide, 186, 137 dye-mediated generation, 186, 638 and ferryl species, distinction, 186, 148 induced DNA damage photodynamic studies, 186, 502 selected-ion MS assays, 186, 530 kinetics and mechanisms, 186, 93 and O- radicals, reaction with polyaminocarboxylates, 186, 117 production by Fenton reaction with Fe-EDTA chelates, 186, 15 superoxide-generating and other systems, 186, 17 role of metal ions, 186, 19 site specificity, 186, 23 reactions in solutions, rate constants, 186, 114 role in human disease, 186, 10 site-specific production, deoxyribose assay, 186, 26 spin trapping, 186, 127 4-Hydroxynonenal GC/MS, 186, 399 HPLC and GC/MS assays, 186, 419 p-Hydroxyphenylacetaldehyde in two-stage labeling of proteins, 182, 734 4-Hydroxyphenylacetate [tTO] or UsO]hydroxyl-enriched, synthesis via diazonium reaction, 188, 108 4-Hydroxyphenylacetate 3-hydroxylase purification from Brevibacterium fuscum, 188, 112 Hydroxypyruvate reductase assay, 188, 373 properties, 188, 377 purification from Methylobacteriurn extorquens AM1,188, 374 8-Hydroxyquinoline coated glass beads, in generation of metal-free solutions, 186, 124 N-Hydroxysuccinimide ester in two-stage labeling of proteins, 182, 733 N-Hydroxysulfosuccinimide sodium salt, synthesis, 172, 622 Hygromycin resistance of yeast, assessment, 185, 479

117 Hygromycin phosphotransferase -human serum albumin fusion cassette, construction, 185,476 Hyperpolarization light-induced, in Nitella demonstration, conditions for, 174, 483 dependence on ATP hydrolysis, 174, 487 electrophysiology, 174, 480 membrane, ouabain-sensitive, in assay of N a t - K + pump activity, 173, 712 Hypersensitivity delayed, in assay of retinoid immunomodulatory activity in vivo, 190,257 retinoid-mediated cell-mediated immunity, 190, 264 Hypertension juxtaglomerular interstitial cell-induced, characteristics, 191, 158 Hypertonic treatment permeabilization of mammalian cells to macromolecules, 171, 868 Hypervitaminosis A testing, 190, 387 Hyphomicrobium

DM2, purification of dichloromethane dehalogenase, 188, 357 X, purification of aldehyde dehydrogenase, 188, 329 cytochromes cn and cL, 188, 305 methanol dehydrogenase, 188, 204 Hypophysectomy in functional analysis of hypothalamohypophyseal axis, 168, 242 Hypothalamic releasing factors microdialysis assay, 168, 168 Hypothalamus human, tissue perifusion in vitro, 168, 206 rat neural transplantation, 168, 247 neuroendocrine mRNAs, nonradioactive visualization, 168, 753 preoptic region and median eminence, proLH-RH extraction, 168, 527 somatostatin release effects of neurotransmitters and neuropeptides, 168, 728

Idiotypic networks in vitro analysis, 168, 719 in vivo analysis, 168, 716

Hypoxanthine -xanthine oxidase, in assay for superoxide dismutase, 186, 227 Hypoxia effect on glutathione defense system, 186, 756 -ischemia, low-flow, and no-flow ischemia, comparative biochemistry, 186, 754 related oxidant stress, analysis in isolated perfused organs, 186, 752

I Idiotopes, see Idiotypic determinants Idiotypes lymphoid tumor, constancy, 178, 348 on monoclonal antibodies, detection by immunoblotting, 178, 144 Idiotypic cascade during schistosomiasis, and isotypic selection of Ab3, relationship, 178, 396 Idiotypic determinants on antiallergen antibodies, assays, 178. 150 interspecies cross-reactive, detection by inhibition ELISA, 178, 384 in lymphoid tumors, 178, 342 paratope-related, detection by inhibition ELISA, 178, 383 topography, 178, 27 Idiotypic networks alprenolol, components, 178, 274 associated mimicry, versus molecular mimicry, 178, 32 bacterial, related epitopes, 178, 12 connectivity, mimicry concept based on. 178, 33 in experimental autoimmune and clinical myasthenia gravis, 178, 441 historical perspectives, 178, 3 and molecular mimicry, overview, 178, 3 parasitic, 178, 13 tumor antigen-related, 178, 15 viral, related epitopes, 178, 8

Idiotypy Idiotypy in autoimmune reactivity to acetylcholine receptor, 178,448 Image analysis anti-idiotype-immunostained receptors in rat brain, 178, 221 autoradiographic signals, 168, 410 in fluorescence microscopy, 173, 752 in measurement of endosomal and lysosomal pH, 173, 757 local cytoplasmic Ca2+ levels, 173, 766 red blood cells in ektacytometry, 173, 39 skin surface impressions, correlation with clinical gradings, 190, 366 Imaging after anti-idiotype immunostaining, rat brain receptors, 178, 219 two-dimensional, chemiluminescence in biological systems, 186, 604 2-Iminobiotin affinity columns, preparation, 184, 169 2-Iminobiotin-6-aminohexyl-Sepharose CL4B preparation, 184, 82 2-Iminobiotin hydrobromide synthesis, 184, 168 Immobilization antisera for immunoaffinity purification of arachidonic acid metabolites, 187, 65 covalent, carbohydrates on polyacrylamide gels, 179, 542 IgGs to Sepharose matrices, 181, 238 non-covalent, glycosphingolipids on plastic microwells, 179, 546 proteins by complex formation with colloidal gold, 179, 120 yeast for immunofluorescence studies, 194, 576 streptavidin, 184, 634 Immune complexes idiotype-anti-idiotype, immunoelectron microscopy, 178, 130 immunogens for anti-idiotype therapy of lymphoid tumors, 178, 351 Immune responses autogeneic, s e e Autoimmune responses cell-mediated, retinoid-mediated assays, 190, 264

118 humoral, assessment, 19tl, 266 local, under vitamin A deficiency, analysis, 190, 272 nonhuman primates to synthetic hepatitis B antigen peptides, 178, 656 polyclonal anti-idiotypic, to substance P antibodies, 178, 279 Immune suppression anti-DNA antibody production with antiId antibody-neocarzinostatinconjugates, 178, 422 Immunity autogeneic, s e e Autoimmunity to poliovirus, canine parvovirus, and rabies virus, induction by antiidiotypic antibodies, 178, 387 Immunization for anti-idiotype therapy of lymphoid tumors immune complexes for, 178, 351 immunogen sources, 178, 349 raising monoclonal anti-idiotypes, 178, 351 with anti-idiotypic antibodies Ab2a or Ab2fl, subsequent Ab3 response, 178, 377 with FMDV peptides with added foreign Th-cell epitopes, 178, 666 peripheral lymphocytes in vitro, 182, 677 in preparation of anti-idiotypic antibodies to morphine, 227 to prolactin, 178, 313 protocol, 178, 45 catalytic antibodies, 178, 540 monoclonal antibodies procedure, 182, 674 to small ligands, 178, 50 polyclonal idiotype for analysis of anti-Id antibodies to hormone receptors, 178, 197 prolactin antibodies, 178, 312 site-specific antipeptide antibodies to rhinovirus receptor, 178, 680 syngeneic monoclonal anti-idiotypic antibodies in viral systems, 178, 382 rabbits for polyclonal antibody preparation, 182, 665 with synthetic peptides, 178, 704

1 19 lmmunoadsorption in analysis of antigenic determinant exposure in nucleosomes, 170, 234 dispersed cells from renal epithelia, 171, 588 double, for reduction of nonspecific binding, 182, 697 EGF receptor, effect of ganglioside Gu~, 179, 525 radiolabeled proteins by S t a p h y l o c o c c u s aureus, 182, 688 vesicles, in analysis of hepatocyte plasma membrane biogenesis, 191, 837 Immunoaffinity chromatography with anti-idiotypic antibodies, nicotinic receptors, 178, 190 anti-idiotypic antibodies to opioid receptors. 178, 245 anti-m~A, snRNPs U2 and U4/U6, 181, 244 anti-m3G, nucleoplasmic snRNPs, 181, 241 arachidonate 5-1ipoxygenase, 187, 338 arachidonic acid metabolites, 187, 62 column preparation, 168, 603; 181, 236 differential, snRNPs U1, U2, and U5, 181, 246 DNA-protein complexes, 170, 398 inhibin, 168, 606 snRNP Sm, 181, 258 transferrin, 184, 301 Immunoassay antibodies after isoelectric focusing and gel blotting, 178, 60 in antibody mapping of sodium channel functional sites, 178, 727, 731 antigen and antibody markers, 182, 705 antihistone antibodies in nucleosome studies, 170, 258 anti-idiotype inhibition, 178, 61 antiserum preparation, 182, 702 avidin-biotin-mediated, overview, 184, 467 benzene dioxygenase, 188, 55 binding, acidic polysaccharides, 179, 95 with charcoal, secretin, 182, 711 chemiluminescent competitive, salivary cortisol, 184, 481 two-site, human growth hormone, 184, 481

Immunoblotting for diagnosis of infectious disease, overview, 184, 529 double-antibody immunoprecipitation, creatine kinase isoenzymes. 182, 712 enzyme, see Enzyme immunoassay HIV core proteins, 184, 556 idiotopes on antiallergen antibodies specific antiallergen IgG antibodies. 178, 155 specific antiallergen IgE antibodies. 178, 158 purified anti-ld antibodies, 178, 162 immunologic cross-reactivity in, reduction, 182, 717 incubation conditions, 182, 706 for monoclonal idiotypes and antiidiotypes for small ligands, selection, 178, 51 in peptide processing, analysis in vivo, 168, 520 proteins, overview, 182,700 radioactive, see Radioimmunoassay repeatability, 182, 713 separation systems, 182, 708 solid-phase with polyacrylamide-antibody beads. human IgA, 182, 709 immunoenzyme, 182, 712 standardization, 182, 713 synthetic peptide, for distinguishing HIV-I and HIV-2 infections. 178, 693 tissue and serum samples, 182, 705 variance, 182, 713 validation, 182, 714 Immunoblotting acidic polysaccharides, 179, 95 in analysis of anti-synthetic peptide antibodies to rhinovirus receptor, 178, 688 keratins, 190, 97 plasma histidine-rich glycoprotein, 169, 273 thrombin-stimulated platelet releasate. 169, 273 antisera analysis in preparation of immunological probes for nucleosomes, 170, 224 colony, in detection of extracellular vacuolar proteins, 194, 660

Immunoblotting in detection of idiotypes on monoclonal antibodies, 178, 144 in enzyme immunoassays with avidinbiotin system, 184, 480 proteins artifacts, 182, 687 procedures, 182, 683 retinoid-bindingproteins, 189, 303 for screening monoclonal antibodies, 182, 674 in testing specificity of anti-receptor anti-idiotype antibodies, 178, 189 in tyrosine monooxygenase assay in median eminence and superior cervical ganglion, 168, 371 vegetative tissue lectins, 179, 342 Immunochromatography epitope-tagged proteins, 194, 517 Immunocytochemicai assay with anti-idiotypic antibodies, receptors, 178, 212 antisubstance P anti-idiotypic antibody binding to physiological receptors, 178, 293 with avidin-biotin system, general procedure, 184, 479 polyclonal anti-idiotypic anti-morphine antibodies, 178, 236 Immunocytochemistry anti-idiotypic antibodies to opioid receptors in hybrid cells, 178, 251 in detection of alcohol oxidase protein in Hansenula polymorpha, 188, 417 human T-cell and B-cell antigens, 184, 363 platelet granule proteins, 169, 301 electron, in detection of diffuse neuroendocrine system components, 168, 799 platelet PADGEM protein, 169, 325 light microscope, plasma membrane proteins, 184, 638 neuroendocrine system binding site analysis, 168, 804 localization of system components, 168, 792 renal epithelia in situ, 191, 279 nucleosomes in analysis of composition and organization, 170, 230

120 application of immunological probes, 170, 234 Immunodissection in isolation of renal epithelia, 171, 584 lmmunodots luminescent detection, 184, 442 lmmunoelectron microscopy in chromosome organization analysis, 170, 238 with gold labeling, yeast ultrathin sections, 194, 608 idiotype-anti-idiotype complexes, 178, 131 nucleosomes antibody preparation and selection for, 170, 181 demonstration of heterogeneity, 170, 239 indirect localization methods, 170, 191 subnucleosomal localization, 170, 184 platelets, in localization of granule proteins, 169, 301 ribonucleoprotein complexes on nascent RNA, 180, 507 Immunoelectrophoresis antibody detection, evaluation, 182, 668 radioactive, see Radioimmunoelectrophoresis Immunoenzymatic assay slide, human immunoglobulin E, 184, 501 Immunofluorescence in analysis of antibody suitability for immunoelectron microscopy, 170, 182 antisubstance P anti-idiotypic antibody binding to physiological receptors, 178, 292 polytene chromosomes, 170, 241 in detection of platelet PADGEM protein, 169, 322 double-label, in yeast studies, 194, 597 in flow cytometry methods, 171, 573 overview, 171,559 and phalloidin staining of actin, in yeast studies, 194, 731 in screening hybridoma cultures for monoclonal antibodies, 171, 583

121 in studies of Schizosaccharomyces pombe, 194, 822 in yeast studies, 194, 567 Immunofluorescence microscopy yeast vacuoles, 194, 649 lmmunofractionation chromatin and nucleosomes, 170, 245 Immunogenicity hepatitis B virus nucleocapsid versus envelope, 178, 650 peptide epitopes, enhanced, peptide vaccines based on, 178, 659 lmmunogens for anti-idiotype therapy of lymphoid tumors, sources, 178, 349 for antibodies to hepatitis B components, synthetic T cell epitopes as, 178, 634 overlapping peptide studies, 178, 626 synthetic peptide, for obtaining antiprotein antisera, selection, 178, 583 Immunoglobulins, see also Antibodies biotinylation, 184, 536 encoding genes cloning, 178, 460 for production of chimeric antibody with antifibrin/t-PA functions, cloning, 178, 517 IgA human, solid-phase immunoassay with polyacrylamide-antibody beads, 182, 709 murine, variable domains, secretion, purification, and dimerization, 182, 138 lgE, slide immunoenzymatic assay, 184, 501 IgG fraction in antisera, isolation, 187, 64 immobilization to Sepharose matrices, 181, 238 synapsin I, immunoprecipitation of bovine synaptic vesicles, 174, 13 thrombospondin, preparation, 169, 261 IgM lectin-bound, in affinity chromatography of oligosaccharides, 179, 56 murine B-cell lymphoma, anti-idiotype preparation for drug conjugation, 178, 370

Immunoprecipitation Immunohistochemistry in analysis of cellular retinol- and retinoic acidbinding protein tissue distribution, 189, 315 vitamin A-regulated keratin expression, 190, 25 with avidin-biotin reagents, description, 184, 357 and in situ hybridization, combination. 168, 771 Immunolabeling double fluorescence antigens, 184, 343 integrin and zeugmatin in embryonic heart cells, 184, 350 talin and vinculin in embryonic fibroblasts, 184, 351 with gold, yeast ultrathin sections, 194. 608 Immunology network hypothesis, historical perspectives, 178, 3 Immunoneutralization active, comparison with passive methods, 168, 499 passive affected biological activity, replacement with modified agonists, 168, 513 in analysis of basal and pulsatile hormone secretion regulation, 168,498 comparison with active methods, 168, 499 interference with radioimmunoassays, 168, 509 validation, 168, 504 Immunopathology in assessment of nutrient immunomodulating effects, 190, 263 Immunoprecipitation arachidonic acid metabolites, 187, 68 bovine synaptic vesicles with antisynapsin I IgG, 174, 13 in chimeric antibody molecule characterization, 178, 472 depletion of snRNPs in nuclear extracts. 181, 83 double-antibody, assay of creatine kinase isoenzymes, 182, 712

Immunoprecipitation epitope°tagged proteins from yeast, 194, 515 erythroid band 3 protein expressed in Xenopus oocytes, 173,457 hepatocyte plasma membrane proteins, 191, 833 low-stringency, RNA, 181, 276 -protection experiments, 180, 458 proteins in detection of degradation in intact cells, 186, 498 expressed in mammalian cells, 185, 582 overview, 182, 688 yeast, in transport analysis, 194, 672 radioactive, see Radioimmunoprecipitation ribonucleoproteins with autoantibodies, 180, 468 RNA, 181, 276 RNA with bound factor after RNase digestion, 180, 456 RNA polymerase II, 194, 517 snRNAs by anti-Sm antiserum, 181, 24 in specificity testing of anti-receptor anti-idiotype antibodies, 178, 189 yeast invertase and chitinase, 194, 694 Immunoprophylaxis schistosomiasis, anti-idiotypic antibodies in, 178, 390, 402 Immunosedimentation in analysis of nucleosome composition, 170, 232 Immunosorbents for analysis of idiotopes on antiallergen antibodies, preparation, 178, 152 Immunostaining anti-idiotype, receptors in rat brain, 178, 216 fluorescent cultured glomerular mesangial cells, 187, 551 in flow cytometry, 171, 573 high-molecular-weight proteins, electrophoretic transfer for, 172, 687 histochemical, with avidin-biotin complex, description, 184, 358 Schizosaccharomyces pombe, 194, 823 ubiquitin-protein conjugates in vivo, 185, 392

122 yeast ultrathin sections, solutions and antibodies for, 194, 617 Immunotoxins anti-idiotypic, in treatment of B-cell neoplasms preparation, 178, 358 in vitro assay, 178, 361 in vivo applications, 178, 363 selective, mutant diphtheria toxinmonoclonal antibody conjugates ADP-ribosylation assay, 178, 414 cytotoxicity assay, 178, 415 inactivation of protein synthesis, kinetics, 178, 418 mutant DT for, preparation, 178, 407 synthesis and purification, 178, 411 in treatment of myasthenia gravis, 178, 451 Impedance in analysis of electrical analog of Necturus gallbladder, 171,653 leaky epithelia, 171, 642 tight epithelia, 171, 628 computational methods with Fourier analysis, 171, 634 current waveform selection, 171, 646 curve-fitting for determination of epithelial circuit parameters, 171, 635 data sampling and analysis, 171, 649 definition, 171, 629 in giant algal cells, measurement with voltage clamp, 174, 427 inhomogeneity problems, 171, 652 instrumentation, 171, 632, 648 leak effects, 171, 657 measurement, 171, 630 principles, 171, 644 statistical analyses, 171, 637 transepithelial gastric mucosa, measurement, 192, 84 measurement, 191, 17 Implantation retinoic acid-impregnated beads in embryonic tissues, 190, 205 Incubation pancreatic minilobules, 191, 686 red cells, media for, in identification of amino acid transporters, 173, 132 retinal pigment epithelial cells

123

Insulin-like growth factor II receptors

with all-trans-[3H]retinol, 190, 16 short-term system, 190, 8 Indanyloxyacetic acids affinity column, synthesis, 191, 576 IAA-94, radiolabeling, 191, 574 Indium- 111 labeling of platelets, 169, 174 lndo-1 in assay of platelet [Ca2+]i, 169, 410 entrapment by mammalian mitochondrial matrix, 174, 115 Infection airway epithelial cells by retrovirus, 192, 568 CV1 cells with simian virus 40, 170, 15, 46 HIV-I and H1V-2, distinguishing immunoassays, 178, 693 viral, MDCK cells, 191, 815 Infectious diseases anti-idiotype antibodies as vaccines, 178, 7 diagnosis with immunoassays, 184, 529 Inflammatory cells counting on chemotaxis membranes, 169, 241 platelet-derived growth factor-induced chemotaxis assay, 169, 236 checkerboard analysis, 169, 241 Influenza virus WSNts61, infection of MDCK cells, 191, 815 Infrared spectroscopy difference, retinal proteins, 189, 123, 130 retinoic acid analogs, 189, 26 Infusion intragastric, etretinate via implanted osmotic minipumps, 190,444 micro-, s e e Microinfusion Inhibin antisera, purified, preparation, 168, 603 bioassay in vitro, 168, 602 concentration from biological fluids, 168, 616 detection and purification, 168, 588 iodination, 168, 596 radioimmunoassay, 168, 590 Western blot analysis, 168, 610

Initiation factors elF-2, mediated translation in expression of cDNA genes in mammalian ceils, 185, 504 Injection micro-, s e e Microinjection Inositol detection in renal extracts by 2D COSY. 176, 336, 493 starvation, in enrichment of yeast mutants, 194, 279 tritiated, loading of hepatocytes, 173, 535 Inositol lipids hydrolysis, 168, 343 metabolism in endocrine cells, analysis by transient permeabilization, 168. 218 Inositol phosphates assay, 168, 342 chromatography, 173, 536 dephosphorylation, 168, 343 HPLC, 173, 537 isomers, radiolabeled, two-stage analysis, 191,707 Inositol phospholipids endogenous platelet, assays, 169, 415 Inositol 1,4,5-trisphosphate in liver, assay, 173, 535 microinjection into cells, 191, 704 phospholipase C-catalyzed production by neutrophil membranes, assay, 187. 232 sensitive stores, and Ca 2~ entry, selective inactivation, 191, 703 Insulin binding assays, 184, 253 biotinylation, 184, 247 dose-response testing by hippocampal brain slice recording, 168, 141 encoding gene, transfection of secretory cells, 192, 227 pro-. s e e Proinsulin Insulin-like growth factor 11 analysis with IGF-1I antibodies, 168, 320 coupled affinity column, preparation, 168, 313 Insulin-like growth factor 1I receptors antibodies applications, 168, 320 assay, 168, 318

Insulin-like growth factor II receptors characteristics, 168, 320 preparation, 168, 318 assay, 168, 312 characterization, 168, 309 localization, 168, 322 purification, 168, 309 regulation, 168, 322 relationship to mannose-6-phosphate receptor, 168, 323 Insulin receptors anti-idiotypic antibodies to, 178, 23 anti-idiotypic anti-DM Id monoclonal antibodies biological properties, 178, 307 production, 178, 303 associated kinase, effect of gangliosides, 179, 534 DM idiotype monoclonal antibodies, production, 178, 306 isolation, 184, 247 properties, 184, 259 Integrin and zeugmatin, double fluorescence immunolabeling in embryonic heart cells, 184, 350 Intensity fluctuation spectroscopy in determination of lipid vesicle size distribution, 172, 383 Intercellular communication analytical technologies, evaluation, 168, 70 in peptide target cells of anterior pituitary, 168, 47 Interfaces air-liquid, culturing of keratinocytes at, 190, 23 protein-lipid, mapping, 172, 631 Interferon-/3 promoter, for expression of cDNA genes in mammalian cells, 185, 497 Internal image in myasthenia gravis, anti-idiotypes as, preparation, 178, 443 role in anti-idiotypic response to antialprenolol antibodies, 178, 274 substance P, anti-SP anti-idiotypic antibodies as, 178, 290 Interphotoreceptor matrix frog, preparation, 189, 326

124

purification and assay of retinoid-binding protein, 189, 213 vertebrate eye, preparation, 189, 215 Intestine duodenum, mucosa, HCO~ secretion, 192, 147 epithelial cells ATP-depleted, in analysis of transport functions, 192, 335 basolateral membrane, Ca 2+ transport, 192, 448 electrochemical gradients of defined magnitude, creation, 192, 337 intracellular Na ÷ and K +, assay, 192, 337 membrane potentials of defined magnitude, creation, 192, 337 measurement, 192, 338 c~-methylglucoside accumulation, 192, 329 Na ÷ gradients of defined magnitude, creation, 192, 336 Na÷-independent sugar transport, 192, 334 transport functions, evaluation, 192, 324 villous, isolation, 192, 324 feline ischemia-reperfusion injury, role of leukocytes, 186, 729 postischemic mucosa, leukocyte infiltration, assay, 186, 729 ischemia-reoxygenation injury, 186, 70 large, s e e Colon membrane vesicles, NaC1 transport pathways, 192, 389 purification of vasoactive intestinal peptide, 168, 305 rat, mRNA preparation, 191, 441 reperfusion injury, role of oxygenderived free radicals, 186, 711 secretory cells intracellular Ca 2÷ assay in single isolated cells, 192, 317 effect on membrane current, 192, 316 membrane K ÷ conductance

125 characterization, 192, 309 pharmacological properties, 192, 316 membrane potential and current measurements, 192, 311 small brush border and basolateral membrane vesicles, transport studies, 172, 346 guinea pig ileum, antagonist activity of anti-Id antibodies to substance P, 178, 297 tissue heterogeneity, resulting membrane vesicle heterogeneity, 172, 356 teleost, ion transport studies, 192,746 tissue, cells from, elutriation, 171, 494 lnvertase, see/3-Fructofuranosidase Iodide reduction of hydroperoxides, 186, 519 lodination cellular retinoic acid-binding protein, 189, 278 cellular retinol-binding protein, 189, 278 histamine, 184, 213 histidine, 184, 217 inhibin chains, synthetic fragments, 168, 594 natural products, 168, 596 serum retinol-binding protein, 189, 272 thrombospondin, 169, 262 Iodine 1251 labeled ligands, binding to washed human platelets, 187, 401 uncoupled, removal and storage in radiolabeling of proteins, 182, 735 radioactive indicator molecules, 182,721 Iodine monochloride in direct iodination of proteins, 182, 729 Iodogen in direct iodination of proteins, 182, 731 radioiodination of proteins for peptide mapping, 182,617 lodometric assay hydroperoxides, 186, 434 Ion channels, see also specific channels apical, analysis in turtle colon, 192, 737 in gall bladder epithelium, identification and characterization, 192, 681

Ion-exchange chromatography in intact erythrocytes, crosslinking with DTSSP, 172, 625 in Malpighian tubule, patch clamp studies, 191, 181 mediated fluxes in membrane vesicles. assay, 172, 155 in pancreatic acinar cells, electrophysiology, 192, 303 Ion currents detection by selected-ion monitoring, 193, 86 in phospholipid vesicles, estimation, 172, 341 role in acinar fluid secretion, 192, 306 single-channel patch-clamp studies: demonstration of messenger-mediated channel opening, 171, 667 recorded in excised membrane patches, examples, 171,665 whole-cell, analysis: demonstration of messenger-mediated channel opening, 171, 667 Ion-exchange chromatography N-acetylglucosaminyltransferase 11. 179, 472 anti-m3G affinity-purified snRNPs. 181, 251 arachidonate 5-1ipoxygenase, 187, 273 bilitranslocase, 174, 55 column packings preparation and regeneration, 182, 3 types of, 182, 310 cytochrome P450 prostaglandin ~0-hydroxylase, 187, 256 formaldehyde dehydrogenase, 188, 319 glutaminylpeptide cyclase product, 168, 361 high-performance gastrin-releasing peptides, 168, 671 oligosaccharides, 193, 517 O-linked oligosaccharides, 179, 22 proteins, 182, 403 with pulsed amperometric detection glycoconjugate monosaccharides, 179, 76 oligosaccharides, 179, 65 histone subunits, 170, 457

Ion-exchange chromatography 4-hydroxyphenylacetate 3-hydroxylase, 188, 113 inositol phosphate isomers, 191, 708 integral membrane proteins, 182, 513 ion exchanger types, 182, 310 low-pressure, gastrin-releasing peptides, 168, 670 mannoprotein, 185, 456 monoclonal anti-idiotypic antibody to opioid receptors, 178, 254 Na÷-n-glucose cotransporter polypeptide components, 191, 603 operation practice, 182, 313 principles, 182, 310 packing preparation and regeneration, 182, 311 parathyroid hormone, 184, 278 polygalacturonic acid oligomers, 179, 568 proteins, overview, 182, 309 protocatechuate 3,4-dioxygenase, 188, 86 pyrroloquinoline quinone, 188, 261 reconstituted vesicles in assay of Ca2+ transport, 172, 41 RNA, 180, 26 sulfated glycosaminoglycans, 179, 433 UI snRNP particles lacking Ul-specific proteins, 181, 254 Ionization chemical Brensted acid reagents, 193, 14 Brensted base reagent ions, 193, 19 charge-exchange reagent systems, 193, 16 electron capture methods, 193, 17 essential reactions, 193, 10 glycosphingolipids, 193, 723 instrumentation, 193, 12 mass reference standards, 193, 309 desorption definition, 193, 20 laser methods, 193, 28 particle beam methods, 193, 23 electron instrumentation, 193, 5 mass reference standards, 193, 305 mass spectra, origin, 193, 6 principles, 193, 3 electrospray

126 atmospheric pressure ion source, 193, 414 in mass spectrometry, 193, 412 in FAB/MS and liquid secondary ion MS, 193, 208 field, mass reference standards, 193, 309 fields and sprays in, 193, 34 glycosphingolipids, 193, 722 -laser desorption, in MS analysis of peptides and proteins, 193, 280 Ionomycin in calibration of quin 2 fluorescence for assay of cytosolic free Ca2+, 172, 246 Ionophores, see also specific ionophores in characterization of epithelial membrane properties, 171, 715 permeabilization of mammalian cells to macromolecules, 171, 863 Ions, see also specific ions Bj from monosaccharidic glycosides, spectral assignment, 193, 610 B2 from disaccharidic glycosides, spectral assignment, 193, 615 B. derived from sugar moieties, analysis, 193, 621 collision-induced decomposition, analysis with magnetic sector instruments, 193, 47 compartmentation in root cells and tissues, X-ray microanalysis, 174, 267 disaccharidic sugar, preparation for tandem MS, 193, 609 distribution in yeast ceils, 174, 608 fluxes in isolated renal cells, assay, 191,403 in membrane vesicles, assay by rapidreaction methods, 172, 301 related to water transport across salivary acini, assay, 192, 33 fragmentation, from FAB/MS, in peptide sequencing, 168, 101 fragments from partially methylated alditol acetates, identification, 193, 565 interactions at membranous polypeptide sites, NMR studies, 171, 286 binding constant determination, 171, 311

127 binding site localization, 171, 335 rate constant determination, 171, 306 intracellular levels in A m p h i u m a erythrocytes, 173, 333 lipophilic, equilibrium distribution, 172, 64 in mass spectra of trimethylsilyl derivatives of nucleosides, 193, 827 metal, see Metal ions molecular, multiply charged, collisioninduced dissociation: analysis of oligopeptides and proteins, 193, 428 paramagnetic, impurities in NMR samples, 176, 79 peptide fragment, nomenclature, 193, 886 permeability of isolated perfused renal tubules, 191, 369 in rapidly frozen cells, electron probe Xray microanalysis, 172, 203 reference, for exact mass measurements, 193, 870 selectivities of gall bladder epithelial cell membranes, determination, 192, 659 lontophoresis in electrophysiological assay of opioid peptides in vivo, 168, 108 Ion transport in airway epithelial cells, normal phenotype persistence, 192, 565 alkali metal ions in Rhodotorula, 174, 645 carrier-mediated analysis with planar bilayers, 171, 276 kinetics, 171, 62 channel-mediated assay, 172, 158 kinetics, 171, 62 in Chara cells, 174, 443 fluxes during electrophysiological transients, 174, 469 kinetic modeling, 174, 475 membrane fluxes, 174, 450 net fluxes, 174, 459 regulating factors, 174, 472 cotransport with molecules, 171, 160 electrogenic and electroneutral transporters, regulation in tracheal epithelium, 192, 549 across energy barriers, analysis, 171, 68

Iron in gall bladder epithelium, electrophysiological studies, 192, 650 across gastric mucosa, electrophysiological studies, 192, 82 mechanisms, analysis with Ba '-*, 171, 621 membrane, assay on individual protoplasts, 174, 322 by one-step inside-out vesicles, assay, 173, 371 in parietal cell membranes, assay, 192, 38 related functions, dependencies on membrane conditions, detection. 171, 225 simple electrodiffusion, 171, 69 stelar, mechanistic analysis, 174, 287 in sweat gland, in vitro analysis, 192, 585 systems, characterization with amiloride and analogs, 191, 739 in teleost intestine electrophysiological studies, 192, 751 transepithelial ion fluxes and dilution potentials, 192, 747 uptake analysis, 192. 750 in yeast determination, 174, 603 kinetics, 174, 606 IPTG, see lsopropyl-fl-D-thiogalactopyranoside Iris rabbit, ginkgolide interactions with platelet-activating factor binding sites, 187, 437,440 Iron complexes for radical reactions, bleomycin assay, 186, 36 - E D T A chelates, role in hydroxyl radical production by Fenton reaction, 186, 15 Fe(IV) and Fe(V), complexes in aqueous solutions, generation, 186, 108 Fenton-reactive, in vivo sources, 186, 37 intracellular quenching of quin2, 172. 251 metabolism in humans, overview, 186, 29 -polyaminocarboxylate complexes, reactions with free radicals, 186, 119

Iron redox reactions and lipid peroxidation, 186, 457 role in lipid peroxidation, 186, 45 simple chelates, role in human disease, 186, 35 Ischemia effect on glutathione defense system, 186, 756 no-flow, and low-flow ischemia-hypoxia, comparative biochemistry, 186, 754 -reoxygenation injury brain, 186, 80 effects on organ preservation for transplantation, 186, 81 heart, 186, 74 intestine, 186, 70 limbs, 186, 84 pancreas, 186, 84 in rheumatoid arthritis, 186, 83 role of endothelium, 186, 78 -reperfusion heart eicosanoid and PAF release and effects, 187, 616 mitochondrial membrane potential during, measurement, 186, 723 intestine, leukocyte involvement during, 186, 729 related oxidant stress in isolated perfused organs, 186, 752 Islet cells

/3 biochemistry of cell lines, 192, 232 clone morphology, 192, 228 cultured, differentiated function, 192, 234 establishment and culture, 192, 223 impalement with microelectrode, 192, 242 membrane potential measurements, 192, 235 morphology in oitro, 192, 221 cell type separation, 192, 203 growth and replication adult cells, 192, 211 monolayer cultures of newborn cells, 192, 213 hormone biosynthesis, 192, 219 hormone release, 192, 217 isolation, 192, 197

128 primary culture fibroblast overgrowth in, 192, 214 monolayers, 192, 206 newborn and fetal cells, 192, 208 optimal glucose levels, 192, 216 viability, 192, 202 Isocitrate dehydrogenase (NAD ÷) in intact mitochondria, Ca2÷-sensitive properties, assays, 174, 107 Isoelectric focusing hydroxypyruvate reductase, 188, 374 monoclonal antibodies, 178, 56 6-phosphogluconate dehydrogenase, 188, 338 preparative ampholyte removal from proteins, 182, 477 in granulated gel beds, 182, 475 with Rotofor cell, 182, 475 proteins, 182, 459 carrier ampholytes, 182, 464 detergent additives, 182, 466 format, 182, 461 gels casting, 182, 463,468 formation and composition, 182, 462 preservation, 182, 473 pH gradients determination, 182, 473 instability, 182, 466 principles, 182, 460 protein band detection, 182, 472 resolution, 182, 465 sample preparation and application, 182, 469 in 2D gel electrophoresis, 182, 443 soluble cytochromes c, 188, 291 fl-thromboglobulin antigens, 169, 227 thrombospondin, 169, 261 Isoflurophate treatment of neutrophil suspensions, 187. 596 Isoleucine and Leu, differentiation in CID spectra, 193,468 spin system identification, 177, 134 Isopotential spin-dry in orientation of synthetic and native membranes, 172, 403

129 Isopropyl-B-D-thiogalactopyranoside control of sporulation in Bacillus subtilis, 185, 225 induction of Escherichia coli BL21(DE3) and HMSI74(DE3) for expression of target DNA, 185, 79 Isotope dilution in estimation of total body vitamin A reserves. 190, 243 Isotopes abundance distributions, calculation, 193, 882 in analysis of transepithelial electrolyte transport, 192, 363 flux in bovine corneal endothelial cells, assay, 192, 580 detection with spin echoes in alanine aminotransferase, 176, 347 creatine kinase, 176, 351 lactate (2-CH) exchange, homonuclear spin-echo experiments. 176, 345 mass and abundance values, 193, 869 positional exchange, with 3~p NMR, 177, 390 radioaclive in analysis of electroneutral ion transporters in tracheal epithelium, 192, 551 with -'8Mg ion, in Mg2+ transport assay in eukaryotic and prokaryotic cells, 173, 546 in monitoring of amino acid movement across red cells, 173, 133 stable, biologically incorporated, assay, 193, 781 translocation profiles, 174, 299 uptake by membrane vesicles, selective amplification, 172, 155 Isotopic labeling retinoid-binding proteins expressed in Escherichia coli, 189, 516 lsotretinoin, see 13-cis-Retinoic acid Isotropic shift paramagnetic metalloproteins, 177, 249 Isotypic selection antianti-idiotype after anti-schistosomal mAb injection, 178, 396 lvermectin and retinol, cross-competition for parasite retinol-binding protein, 189, 352

Keyhole-limpet hemocyanin J J splitting ~3C-~3C, in analysis of relative fluxes through Krebs cycle and into glucose. 177, 426

K Kadsurenone and related lignans, as platelet-activating factor antagonists, 187, 446 Keratin differentiation marker for embryonal carcinoma cells, 189, 345 gene expression during vitamin A deficiency of tracheal epithelium, 190, 95 vitamin A-regulated shifts, 190, 24 immunoblot analysis, 190, 97 immunolocalization, 190, 24 mRNA, localization, 190, 27 synthesis in tracheal epithelium, role of retinoids, 190.91 Keratinization disorders human, retinoid treatment: murine papilloma test, 190, 382 Keratinocytes cell culture. 190, 31 human culturing at air-liquid interface. 190. 23 differentiation, vitamin A-mediated regulation. 190, 18 gene transfection, 190, 29 keratin expression, vitamin A-regulated shifts, 190, 24 submerged cultures, 190, 20 lipids, retinoid-induced changes, 190, 30 retinoic acid formation from retinol and retinal, 189. 530 Keto groups t80-labeling by acid-catalyzed exchange with H2tsO, 193, 343 Keyhole limpet hemocyanin antibody coupling, for alum precipitation. 178, 46 chemical coupling of synthetic rhinovirus receptor peptides to, 178. 677 coupling of peptides to. 178, 701

Kidney Kidney arterioles blood collection, 191, 121 resistances, 191, 125 artificial, loading and care, 172, 46 blood-free perfusion, 191, 131 bovine, membrane preparation, 191, 574 cell membranes amplification, 191, 269 recycling, 191, 271 cells from functional morphology in situ, 191, 265 isolation, 191, 381 isolated characterization, 191, 391 transport-related parameters, 191, 399 mitochondrial density and transport rate, 191, 272 pH gradient measurements, 191, 470 CI- transport, effects of furosemiderelated diuretics, 191, 785 collecting tubule cells, isolation, 191, 413 cortex cell isolation, 191, 381 purification of basolateral plasma membranes, 191, 456 brush border membranes, 191, 451 vesicles, ATP-driven H ÷ transport, 191, 505 distal tubule cell separation by free-flow electrophoresis, 171, 525 cortical segments, cell isolation, 191, 381 fluid collection, 191, 119 microperfusion studies in vitro, 191, 220 double-microperfusion in situ, 191, 98 drug-dependent H202 generation, assay in vitro, 186, 625 epithelium analysis in situ, 191, 273 cell dispersions, preparation, 171, 586 isolated cells, characterization, 171, 588 isolation with cell-specific monoclonal antibodies, 171, 581

130 mammalian, preparation for study of cell volume regulation, 171, 783 quantitative evaluation, 191, 281 segmental transport functions in vitro, 191,418 frog isolation, 192, 713 perfusion, 192, 713 glomerulus cell isolation and analysis, 191, 141 contraction, determination, 191, 138 filtration rate assessment, 191, 116 mathematical model, 191, 108 isolated incubation for assay of prostaglandin synthesis, 191, 135 incubation for fluid transport studies, 191, 128 microperfusion, 191, 126 preparation and analysis, 191, 130 superfusion system, 191, 136 isolation from rat, 191, 131 juxtamedullary, microperfusion, 191, 126 mesangium, structure and function, 187, 544 micropuncture, 191, 75 glomerular, proximal NaCI and fluid secretion, 191, 195 permselectivity, 191, 49 transcapillary fluid transport, analysis, 191, 107 Henle's loop, thick ascending limb cell isolation, 187, 365; 191, 412 isolated and cultured cells, arachidonic acid metabolism, 187, 369 perfusion, effects of furosemiderelated diuretics, 191, 791 plasma membrane vesicles, isolation, 191, 461 stimulated release of arachidonic acid metabolites, 187, 371 isolated, perfused and nonfiltering, description, 191, 31 juxtaglomerular interstitial cells angiotensin-renin system, detection, 191, 156 induced hypertension, characteristics, 191, 158

131 isolation and culture, 191, 152 kinetics, permeation, and transport in vivo, multiple indicator dilution studies, 191, 34 K + transport, effects of furosemiderelated diuretics, 191, 785 medulla cell isolation, 191, 387 inner, isolation of plasma membrane vesicles, 191, 465 membrane vesicles, ion channelmediated fluxes, assay, 172, 162 micropuncture, 191, 72 Na + transport, effects of furosemiderelated diuretics, 191, 785 nephron segments acid intracellular compartments, density and distribution, 191, 258 binding of hormones and drugs, microassays, 191, 315 cellular pH, measurement, 191, 261 defined isolation and perfusion, 191, 254 transport in isolated ceils, 191, 380 isolated, identification and analysis of specific cell types, 191, 253 microdissected, receptor studies, 191, 306 microdissection, 191, 308 peritubular capillaries, permselectivity, 191, 55 preservation for transplantation, effect of ischemia-reoxygenation injury, 186, 81 proximal tubule cell separation by free-flow electrophoresis, 171, 525 endocytosis of proteins, 191, 340 fluid collection, 191, 117, 120 isolation of cells, 191, 381,410 lysosomal protein hydrolysis, 191, 340 membrane vesicles, transport studies, 172, 346 perfusion, 191, 344 purification of lumenal membranes, 191, 451 reptilian, microperfusion studies in vitro, 191, 203 shark, CI- secretion, 191, 184 tissue heterogeneity, resulting mem-

Kidney brane vesicle heterogeneity, 172, 357 transport and receptor kinetics, 191, 57 renomedullary cells, isolation, 187, 365 renomeduUary interstitial cells, isolation and culture, 191, 162 reperfusion injury, role of oxygenderived free radicals, 186, 715 transplantation, free radical-mediated reperfusion injury during, assay, 186, 748 tubules amphibian functions, 191, 196 isolation and in vitro microperfusion, 191, 197 avian functions, 191, 223 isolation and perfusion studies, 191. 224 elasmobranch functions, 191, 182 isolation and perfusion, 191, 184 functional morphology in situ. 191. 265 fusion, 192,725 intact, water permeability, optical measurement. 191, 565 isolated hormonal receptors, detection and analysis, 191, 303 volume and shape changes, measurements, 191, 371 isolated cells of defined segmental origin electrophysiology, 191, 424 intracellular signal systems, 191,421 primary culture preparation, 191. 409 solute transport and metabolism. 191, 418 isolated, perfused electrophysiology, 191, 289 flux measurements, 191, 354 intracellular solute levels, assay. 191, 366 micropuncture, 191, 78 reptilian functions, 191. 198

Kidney isolation, 191, 199 segments defined, isolation, 191, 326 isolated, metabolism, 191, 325 isolated perfused, transmural water flow measurement, 191, 232 localization, 192, 715 microdissection, 191, 226 recognition, histotopographical criteria, 191, 287 teleost analysis in vitro, 191, 188 functions, 191, 186 isolation, 191, 187 transepithelial electrolyte transport, structural basis, 191, 266 Xenopus, cultured cell-free transcription extracts efficiencies, 170, 355 preparation, 170, 351 nuclei and chromatin, preparation, 170, 348 Kinetics alkali metal/proton exchange, 173, 337, 344 amiloride-blocked Na ÷ uptake by bladder membrane vesicles, 172, 161 amino acid flux in red cells, 173, 138, 155 amino acid transport: recognition and resolution of heterogeneous systems, 173, 576 antibody-epitope interactions, size exclusion HPLC analysis, 178, 114 ATP-driven chloride pump in Acetabularia, 174, 498 biotinyl ligand displacement, 184, 64 carrier-linked transport, 171, 14 cotransport of carbohydrate and H ÷ in Rhodotorula, 174, 631 data from continuous fluorescence monitoring of transport, analysis, 172, 130 enzyme, 2D NMR studies, 176, 330 epoxide hydrolase, 187, 331 fatty acid biosynthesis, ~3C NMR analysis, 177, 429 glutathione transport, 173, 528, 529 inhibition, carrier systems, 171, 113 ion fluxes in Chara cells, 174, 475

132 ion movement mediated by carriers and channels, 171, 62 ion transport in yeast, 174, 606 leukotriene A4 hydrolase, 187, 291,332 membrane proteins, modulation by surface potential, 171, 392 membrane transport, 171, 23 monocarboxylate transport in red cells, 173, 300 Na+/H ÷ exchange of human red cells, 173, 176 nucleoside transporter from red cells, 173, 254 oxygen radicals, 186, 93 protein synthesis inactivation by monoclonal antibody-mutant diphtheria toxin, 178, 418 receptor, in renal proximal tubule, 191, 57 1l-cis-retinaldehyde binding to cellular retinaldehyde-binding protein from pigment epithelium, 189, 373 simple carrier molecules, 173, 149 solute efflux from erythrocyte ghosts, 173, 360 sugar transport in normal and mutant yeast, 174, 621 surface dilution, platelet-activating factor acetylhydrolase, 187, 350 transmembrane equilibration, 172, 339 transport analysis in vesicles, advantages and limitations, 192, 409 computer analysis, 172, 262 in isolated membrane vesicles, 172, 320 in renal proximal tubule, 191, 57 in yeast, 174, 567 uridine transport across red cell membranes, 173, 260 Kininogen high-molecular-weight antisera antibodies, preparation and characterization, 169, 280 coagulant assay, 169, 278 competitive ELISA, 169, 281 platelet form expression on activated platelet surface, 169, 290 function, 169, 295

133

LC/MS

identification, 169, 284 secretion, 169, 286 total in lysed platelets, assay, 169, 286 purification, 169, 279 radiolabeling, 169, 279 Krebs cycle pyruvate entry, ~sC NMR studies, 177, 423 Kupffer cells and hepatic endothelial cells, separation, 190, 56

L Labeling affinity, s e e Affinity labeling with avidin, advantages, 184, 404 blotted antigens and receptors, 184, 424 cells for flow cytometric analysis and sorting, 171, 572 choleragen with biotin, 184, 405 direct, blotted glycoconjugates, 184, 427 DNA with biotinylated nucleotides, 184, 561 end, indirect, in analysis of nucleosome positioning, 170, 408 epoxy- and dihydroxyeicosatrienoic acids with ~80, 187, 184 fluorescent, s e e Fluorescent labeling histone H2A-H2B dimers with gold, 170, 482 MDCK cells with N-6(7-nitro-2,1,3benzoxadiazol-4-yl) ceramide, 191, 816 membranes by monocarboxylate transport inhibitors, 173, 326 metabolic, complex carbohydrates by radiolabeled hexosamines, 179,435 molecules by exchange with HfsO, 193, 338 nucleic acids with photobiotin acetate, 184, 593 photoaffinity, s e e Photoaffinity labeling photochemical, s e e Photochemical labeling proteins and mutants, for conformational comparison, 177, 74, 282 for peptide mapping, 182, 616

pulse, s e e Pulse labeling radioactive, s e e Radiolabeling RNA at 5' and 3' ends, 180, 193 fragments at 5' and 3' ends, 181, 586 with vaccinia capping enzyme, 181. 179 subendothelial extracellular matrix with sulfate, 169, 86 T4 lysozyme, 177, 60 in ultrastructural localization of retinal photoreceptor proteins, 184, 375 Laboratories setup for protein purification, 182. 19 Lactate 2-CH exchange, spin-echo studies, 176, 345 Lactate dehydrogenase cytosol marker, assay, 182, 220 proton-exchange experiments, 176, 343 release by permeabilized parietal cells, assay, 192, 112 Lactoferrin role in iron metabolism, 186, 32 Lactoperoxidase in direct iodination of proteins, 182, 726 generation of cysteine and glutathione thiyl radicals, 186, 324 Lactosamine hexasaccharide alditols collison-induced dissociation spectra, 193, 698 Lactose transport in E s c h e r i c h i a c o l i , characterization. 171, 12 Laminin fibroblast attachment, enhancement by retinoic acid, 190, 87 Langmuir-Blodgett films in orientation of synthetic and native membranes, 172, 407 Laser desorption-ionization with infrared lasers, 193, 28 matrix-assisted methods, 193.29 in MS analysis of peptides and proteins, 193, 280 Lasers in introduction of DNA into mammalian cells, 185, 536 LC/MS, s e e Liquid chromatography-mass spectrometry

Leaves Leaves blender-minced, preparation of guard cell protoplasts, 174, 320 cells and subcellular compartments, metabolite levels, 174, 518 lyophilized and ultrasonicated material, nonaqueous fractionation, 174, 536 maize intercellular fractionation, 174, 543 protoplast isolation, 181, 153 sucrose loading, 174, 293 tobacco, protoplast isolation, 181, 153 wheat mesophyll cells, isolation of deoxyribonucleoprotein, 170, 441 protoplasts with short quench times, subcellular fractionation, 174, 526 Lecithin-retinol acyltransferase assay, 189, 450 in bovine retinal pigment epithelial microsomes, assay, 190, 156 properties, 189, 456 Lectin receptors localization, 184, 388 Lectins Allornyrina dichotoma

-agarose in affinity chromatography of oligosaccharides, 179, 335 preparation, 179, 334 assay, 179, 331 properties, 179, 333 purification from hemolymph, 179, 332 Dolichos biflorus

assay, 179, 342 properties, 179, 346 purification from vegetative tissue, 179, 344 endogenous, prevention of nonspecific binding of avidin, 184, 203 histochemistry, 184, 361 in isolation of cell surface glycoproteins, 184, 304 peanut, binding by cells of isolated nephron segments, 191, 260 snowdrop assay, 179, 327 preparation, 179,328 properties, 179, 329

134 Leech isolation of ceramide glycanase, 179, 481 Lens calf, nuclear homogenates rotational correlation times, 176, 414 translational motion measurement, 176, 439 Leucine and Ile, differentiation in CID spectra, 193, 468 spin system identification, 177, 134 Leukocytes adherence, intravital microscopic assessment, 186, 733 arachidonate 5-1ipoxygenase, isolation and characterization, 187, 312 extravasation, intravital microscopic assessment, 186, 733 functional manipulation in vivo, 186, 739 human DNA strand breaks and repair, analysis, 186, 552 high-molecular-weight DNA, isolation, 194, 255 isolation, 186, 552; 187, 314 respiratory burst, stimulation, 186, 552 infltration of postischemic intestinal mucosa, measurement, 186, 729 5-1ipoxygenase stimulatory factors, preparation, 187, 316 polymorphonuclear, see Neutrophils porcine cytosol fraction, preparation, 187, 340 immunoaffinity purification of arachidonate 5-1ipoxygenase, 187, 338 preparation, 184, 430 role in intestinal ischemia-reperfusion injury, 186, 729 microvascular injury, 186, 736 subsets, simultaneous analysis by flow cytometry, 171, 576 surface proteins, identification, 184, 429 Leukotriene A4 albumin-bound, metabolism into leukotriene C4 by mast cells, 187, 575 hydrolysis hepatic enzymes in, purification and properties, 187, 324

135 product characterization, 187, 325 metabolism, 187, 565 release and metabolism in neutrophilmast cell interactions, 187, 567 Leukotriene A4 hydrolase assay, 187, 289 cDNA cloning, 187, 486 kinetics, 187, 291,332 physical properties, 187, 290 purification from guinea pig liver cytosol, 187, 328 human lung, 187, 286 subcellular distribution in hepatic tissue, 187, 327 substrate specificity, 187, 327 Leukotriene A4 methyl ester hydrolysis, 187, 308 preparation and purification, 187, 287 saponification, 187, 288, 557 tetradeuterated, preparation, 187, 70 Leukotriene B4 assay, 187, 557 formation from leukotriene A4, catalyzing enzymes, purification and properties, 187, 324 GC-NCI-MS analysis, 187, 76 immunogens, preparation, 187, 121 inactivation, 187, 319 metabolism by isolated rat hepatocytes, 187, 285 origin, verification, 187, 557 synthesis by erythrocyte-neutrophil interactions, 187, 553 Leukotriene C4 biosynthesis during neutrophil-vascular cell interaction, 187, 559 enzyme immunoassay with acetylcholinesterase, 187, 82 formation by mast cells from albuminbound leukotriene A4, 187, 575 immunogens, preparation, 187, 118 radioimmunoassay, 187, 563 Leukotriene C4 synthase assay, 187, 336 murine mastocytoma cell assay, 187, 307 characterization, 187, 306 preparation, 187, 310 properties, 187, 337

Light purification from guinea pig lung, 187, 335 Leukotriene D4 immunogens, preparation, 187, 120 Leukotriene D4 receptors antagonists, assessment, 187, 414 Leukotriene E4 enzyme immunoassay with acetylcholinesterase, 187, 82 metabolism by isolated rat hepatocytes. 187, 283 Leukotrienes antibodies, preparation, 187, 116 effects on isolated perfused mammalian heart, 187, 613 metabolism by isolated rat hepatocytes, 187, 277 metabolites, generation and purification, 187, 280 nomenclature, 187, 1 radioimmunoassay, 187, 123 sulfidopeptide in biological fluids, GC/MS assay, 187, 90 in mast cells, assay, 187, 97 LFASTA program in evaluation of local sequence similarity, 183, 88 sequence similarities, 183, 85 LH, s e e Luteinizing hormone LH-RH, s e e Luteinizing hormone-releasing hormone Ligation chromatin after cutting with restriction endonuclease, 170, 12 yeast artificial chromosome vectors, in preparation of clone libraries, 194, 259 Light damaged skin, effects of retinoids, 190, 352 generating reactions, analysis, 186, 604 induced hyperpolarization in N i t e l l a , analysis, 174, 479 for photoaffinity crosslinking of RNA and protein, source selection, 180, 393 signals from calcium-regulated photoproteins

Light calibration, 172, 199 detection, 172, 181 interpretation, 172, 197 ultraviolet, see Ultraviolet light Light adaptation in analysis of retinoid binding by interstitial retinol-binding protein, 189, 326 Light microscopy in analysis of antisubstance P antiidiotypic antibodies, 178, 292 in detection of biotinylated gonadotropin-releasing hormone, 184, 397 in evaluation of carbohydrate receptor binding by colloidal gold, 179, 117 in immunocytochemical localization of plasma membrane proteins, 184, 638 yeast nuclei, 194, 129 yeast nucleolus, 194, 717 Light scattering in analysis of transport in erythrocytes applications, 173, 268 instrumentation, 173, 264 in determination of liposome particle size, 171, 216 dynamic laser, in determination of vesicle size distribution, 172, 364 fluctuations of scattered light, measurement, 172, 373 theory, 172, 367 Lignans kadsurenone-related, as platelet-activating factor antagonists, 187, 446 Lignin peroxidase assay, 1118, 160 production, 188, 164 properties, 188, 168 purification from Phanerochaete chrysosporiurn, 188, 166 Limb bud cell cultures, 190, 422 micromass cell cultures, in testing of retinoid teratogenicity in vitro, 190, 427 Limbs amphibian amputation, 1911, 189 regenerating axes, effects of retinoic acid, 19tl, 191

136 blasterma cell affinity along, position-dependent differences, 190, 198 ischemia-reoxygenation injury, 186, 84 Lindberg method for linkage analysis of carbohydrates, 193, 554 Linear prediction methods comparison with other non-Fourier transform methods, 176, 228 for NMR spectral estimates, 176, 222 Linoleic acid hydroperoxide HPLC, 186, 308 Lipid bilayers chromanol topography and mobility, 186, 367 electrical characterization, 171, 239 interaction with diheptanoylphosphatidylcholine, 171, 272 photochemical reactions in, 172, 646 planar, ion carriers, relaxation and noise analyses, 171, 274 Lipid coumarin dyes as probes of interfacial electrical potential in biomembranes, 171, 376 Lipid hydroperoxides HPLC, 186, 308 HPLC-isoluminal chemiluminescence detection, 186, 371 total, assay with diphenyl-l-pyrenylphosphine, 186, 158 Lipid hydroxy acids GC/MS assay, 186, 683 Lipid mediators electron-capture negative-ion chemical ionization MS, 187, 13 Lipid peroxidation analysis: iron redox reactions, 186, 457 assessment by assay of malondialdehyde, 186, 421 biomedical implications, 186, 55 enzymatic and nonenzymatic, overview, 186, 51 initiation and propagation, 186, 43 microsomal, inhibition by 13-cis-retinoic acid, 190, 281 products formed during oxidant stress in vivo, analysis, 186, 683 relationship to chemiluminescence, 186, 609 role of iron, 186, 45

137 Lipid peroxides extraction from tissues, 186, 389 GC/MS assays, 186, 388 in human body fluids, assays, 186, 66 transesterification to methyl esters, 186, 390 pentafluorobenzyl esters, 186, 391 Lipids adipocyte, binding proteins, purification from human and mouse, 189, 363 association with human tracheobronchial mucin, 179, 17 bilayers, s e e Lipid bilayers in DNA transfection of mammalian cells, 185, 533 dry films, hydration, 171, 198 epidermal composition, 190, 39 effects of retinoic acid, 190, 39 extraction, 190, 32 retinoid-induced changes in keratinocytes, 190, 30 erythrocyte, analysis, 173, 227 extraction, 187, 592 fluorescent probes, visual colocalization in liposomes, 171, 850 in glucose transporter preparations, analysis, 174, 43 high levels, protein assay in presence of, 172, 393 high-performance TLC, 190, 32 hydration, 171, 195 -lipid interactions, analysis, 172, 632 for liposome preparation, selection, 171, 210 membrane core, labeling in intact erythroyctes, 172, 659 reagent requirements, 172, 633 membrane, HPLC, 172, 538 metabolites, production in activated bone marrow-derived mast cells, 187, 517 neutral chromatography, 191,687 extraction from pancreatic tissue, 191, 686 peroxidation, s e e Lipid peroxidation planar, -protein membranes, formation, 171,225 -protein interactions, analysis, 172, 632 -protein interface, mapping, 172, 631

Liposomes -protein vesicles, reconstituted, properties, 171, 263 retinal, photooxidized, GC/MS assay, 186, 393 retinoid binding, 189, 366 solutions, injection in solvent for preparation of large unilamellar vesicles, 171, 207 small unilamellar vesicles, 171, 204 tissue, extraction, 187, 197 vesicles preparation, 171, 255 reconstitution of bacteriorhodopsin into, 171, 267 as retinoid carrier, preparation, 189, 404 retinoid exchange with rod outer segment membranes, 189, 402 Lipoamide dehydrogenase, s e e Dihydrolipoamide dehydrogenase Lipofection introduction of DNA into mammalian cells, 185, 534 Lipooligosaccharides trehalose-containing, purification from mycobacteria, 179, 225 Lipoprotein lipase role in arachidonate liberation from phospholipids, 191, 677 Lipoproteins association with platelet-activating factor acetylhydrolase, 187, 348 effects on platelet-activating factor acetylhydrolase, 187, 351 low-density, isolation, 187, 352 Liposomes added during freeze-thaw reconstitution, formation, 191, 595 fluorescent lipid probes in, resonance energy transfer microscopy, 171, 85O macroscopic, preparation for resonance energy transfer microscopy, 171, 851 32Pi, assay, 173, 738 preparation, 171, 193; 173, 735 size characterization, 171, 193 transfer of retinot from retinol-binding protein to, 189, 394 unilamellar, preparation for spectrophotofluorimetry, 171,851

Lipoxins Lipoxins HPLC, 187, 167 Lipoxygenase generated products of arachidonic acid, HPLC, 187, 98 in oxidation of tocopherols, 186, 203 products, HPLC, 187, 570 reaction with eicosanoids, chiral analysis, 187, 194 Liquid chromatography in assay for retinol, a-tocopherol, and/3carotene in serum. 189, 172 fast protein leukotriene A4 hydrolase, 187, 289 Na+-dependent glucose transporter, 192, 442 in vitamin detection, 189, 179 Liquid chromatography-mass spectrometry continuous-flow fast atom bombardment, peptides and proteins, 193, 225 long-chain bases from membrane lipids, 172, 567 membrane lipids, 172, 565 particle beam interfaces, 193, 122 phospholipids, 172, 570 RNA hydrolysates, 193, 796 thermospray methods, 193, 108 universal interface, 193, 124 Liquids -air interface, culturing of keratinocytes at, 190, 23 Liquid scintillation counting in analysis of erythrocyte anion fluxes, samples for, preparation, 173, 62 Lithium efflux from red cells, assay, 173, 187 Li÷/Na + and Na+/Li + exchanges of human red cells, assay, 173, 184 loading of red cells, 173, 186 Lithium acetate in Schizosaccharomyces pombe transformation, 194, 812 in yeast transformation, 185, 284; 194, 186 Lithium chloride in Schizosaccharomyces pombe transformation, 194, 812 Liver, see also Hepatocytes bile acid transport, mechanisms, 192, 517

138 cell separation and enrichment, 171, 495 glutathione defense system effects of hypoxia or ischemia, 186, 756 properties, 186, 752 guinea pig, purification of cytosolic leukotriene A4 hydrolase, 187, 328 homogenization and subfractionation, 174, 54; 187, 325 myo-inositol 1,4,5-trisphosphate levels, assay, 173, 535 murine cytosol, purification of epoxide hydrolase, 187, 328 inflammatory injury, biochemical pharmacology, 186, 675 perfused drug-induced oxidant stress, assay, 186, 759 in studies of biological transport processes, 192, 485 perfusion, 173, 564 plasma membrane isolation of bilitranslocase, 174, 50 vesicles, alanine transport, 174, 36 preservation for transplantation, effect of ischemia-reoxygenation injury, 186, 81 purification of cytosolic enzymes catalyzing leukotriene A4 hydrolysis, 187, 324 rabbit microsomal membranes, isolation and solubilization, 182, 231 purification of N-acetylglucosaminyltransferases, 179, 364, 369 rat cell types isolation, purification, and characterization, 190, 49 role in retinoid metabolism, 190, 60 drug-dependent H2Oz generation. assay in vitro, 186, 625 endoplasmic reticulum, intactness, measurement, 174, 58 enzymatic perfusion, 191), 62 Golgi membranes, purification of mannosidase IA, 179, 446 Golgi vesicles isolation, 182, 229 preparation and storage, 179, 418

139 mitochondria, purification of phosphate transporter, 173, 732 nonparenchymal cells, density-gradient centrifugation, 190, 66 nuclei, active, preparation, 181, 34 perfusion, 192, 488 plasma membranes, isolation. 182, 227 preparation, 192, 488 purification of DT-diaphorase, 186, 290 polyadenylate-binding proteins, 181, 340 sialate 9-O-acetylesterase, 179, 412 total cell suspension, differential centrifugation, 190, 65 reperfusion injury, rote of oxygenderived free radicals, 186, 715 subcellular distribution of epoxide hydrolase and leukotriene A4 hydrolase, 187, 327 transport analysis, plasma membrane vesicles for, 192, 534 unidirectional Ca 2* fluxes, assay, 192. 495 vitamin A levels relative dose-response test for insufficient stores, 190, 249 static measures, 190, 244 Lobectomy posterior pituitary, in functional analysis of hypothalamohypophyseal axis, 168, 240 Long-chain-fatty-acid-CoA ligase and arachidonate-CoA ligase, separation, 187, 240 Low-angle light-scattering photometry laser, in determination of membrane protein molecular weight, 172, 514 Lowe analysis thermodynamic effects on amino acid transport in red cells, 173, 158 Lowry assay for proteins, overview, 182, 57 LRNA program prediction of optimal and suboptimal RNA secondary structures, 183, 282 Lucerne transient streak virus virusoid cDNA clone, preparation, 181, 584 RNA, self-cleavage, characterization, 181. 583

Lung Luciferin analogs, chemiluminescence, in assay for superoxide dismutase, 186. 227 superoxide radicals and singlet oxygen, 186, 585 Lucifer Yellow yeast cell-associated assay, 194. 701 visualization, 194, 700 in yeast endocytosis assays, 194. 697 L u r n b r i c u s terrestris, s e e Earthworm Luminescence firefly, in assay of ADP and ATP secreted by platelets, 169. 200 Luminol in detection of immunodots and Western blots. 184, 442 Luminometry enzymes, substrates, and pathways in isolated nephron segments. 191. 338 Lung cell separation and enrichment. 171. 495 drug-dependent H20., generation, assay in vitro, 186. 625 guinea pig parenchyma, preparation. 187, 625 purification of leukotriene C4 synthase, 187, 335 whole organ preparation, 187. 625 human bronchial tissue, preparation, 187, 623 from cigarette smokers, preparation and ESR analysis, 186,665 eicosanoid-producing parenchyma. preparation, 187,621 mucus, isolation of tracheobronchial mucin, 179, 6 purification of leukotriene A4 hydrolase, 187. 286 membranes, preparation, 187, 416 rabbit, purification of cytochrome P450 prostaglandin o)-hydroxylase, 187, 253 rat isolated perfused, arachidonate studies, 187, 599 isolation and perfusion, 187, 600 vasoregulation, role of eicosanoids, 187. 607

Luteinizing hormone Luteinizing hormone gonadotropin-releasing hormone-stimulated release from pituitary cells, role of protein kinase C, 168, 287 Luteinizing hormone-releasing hormone precursor processing, analysis, 168, 524, 532 Luteinizing hormone-releasing hormone receptors antibodies, production with complementary peptides, 168, 25 Luteovirus satellite RNAs, self-cleaving sequences, compilation, 180, 547 Lycopene and other nutrients in serum or plasma, simultaneous HPLC, 189, 155 Lyco persicon e s c u l e n t u m , see Tomato Lymphoblastoid cells cytoplasmic RNA, isolation, 180, 8 Lymphoblasts whole-cell suspensions, amino acid loading of lysosomes, 174, 157 Lymphocytes arachidonic acid metabolism, 187, 582 B, see B cells collection from human blood, 186, 228 cytoplasmic RNA, isolation, 180, 8 detection of prostacyclin synthase, 187, 583 effect on platelet function, 187, 584 human anti-DNA idiotype-positive determination, 178, 427 selective elimination, 178, 428 hexose transport, assay, 173, 618 separation by free-flow electrophoresis, 171, 519 immunoselective cell separation, 184, 314 murine, separation by free-flow electrophoresis, 171, 517 Na+/H + exchange, detection, 173, 777 peripheral, immunization and fusion in vitro, 182,677 -platelet interactions in prostacyclin biosynthesis, 187, 578 preparation, 187, 578 proliferation, in assay of retinoid-mediated cell-mediated immunity, 190, 265

140 rat, isolation from thymus, 173, 778 rosette formation, 184, 318 swelling, Na+/H + exchange-induced, detection, 173, 779 T, see T cells Lymphoid cells tumors involving, treatment with antiidiotype antibodies, 178, 341 Lymphokines production and response, effects of vitamin A, 190, 273 Lyophilization leaf material, 174, 537 photoproteins, 172, 170 in preparation of NMR samples, 176, 82, 89 in protein concentration and solute removal, 182, 77 Lysine chromatin DNA-binding residues, radiolaheling, 170, 369 and Gin, differentiation in CID spectra, 193, 468 spin system identification, 177, 140 Lysis auto-, see Autolysis enzymatic gram-negative bacteria, 182, 148 gram-positive bacteria, 182, 147 yeast cells, 182, 161, 164 erythrocyte membranes, 173, 29 erythrocytes from A m p h i u m a , 173, 331 for preparation of resealable white ghosts, 173, 352 higher eukaryotic cells, 182, 195 hypotonic, in preparation of nuclei, 180. II mechanical prokaryotes, 182, 151 yeast cells, 182, 160, 162 nuclei, 180, 12 photo-, see Photolysis Lysolecithin permeabilization of mammalian cells to macromolecules, 171, 866 Lysophosphatidylinositol disposition on stimulation of phosphatidylinositol breakdown, 191, 690 Lysosomal a-D-mannosidase assay, 179, 500

141

Maize

properties, 179, 503 purification from rat epididymis, 179, 501 Lysosomes cystine exodus, analysis, 174, 154 isolated, pH and proton pumping, assay, 174, 146 isolation from mammalian cell lines, 182, 209 marker enzymes/3-hexosaminidase and /3-galactosidase, assays, 182, 215 papaya, purification, 182, 335 pH determination in cell culture, 174, 140 measurement methods, 173, 755 platelet, secretion of acid glycosidase~. assay, 169, 336 proton pumping, assay, 174, 131 in proximal tubules, hydrolysis of proteins. 191, 340 Lysozyme phage T4 assembly details, 177, 226 expression system, 177, 46 first domain structure, solution, 177, 239 forbidden-echo spectra, 177.66 intermediate accessible volumes. analysis, 177, 229 isotopic enrichment, determination, 177.61 ~SN-labeled, NMR, 177, 65 I D and 2D isotope-directed NOE measurements, 177, 69 production from bacterial hosts bearing priSe plasmids, 177, 72 selective ~SN labeling, 177, 60 rotational correlation times, determination by 13C NMR, 176,414 translational motion, measurement by pulse-gradient spin-echo NMR, 176, 439

M Macroelectrodes for measurement of transmembrane potentials and ion activities, 172, 136

Macromolecules biological CODATA Task Group on, 183.58 and components, database listing, 183, 54 on intact erythrocyte membranes. crosslinking, 184, 158 mammalian cell permeabilization to, 171. 857 NMR studies, paramagnetic probes for, 177, 403 permeabilization of cells to. cell cracking method, 168, 225 proton relaxation. 176, 363 rotating-frame NOE. 176. 207 sieving data, in assessment of singlenephron glomerular filtration, 191, 128 systems, conformational exchange in. 176, 371 Macrophages HL60 cell differentiation into. induction by gangliosides, 179, 538 partial purification of phospholipase A2. 187, 223 rat. peritoneal, isolation of galactose/N-acetylgalactosaminebinding protein, 179, 324

mannose/L-fucose/N-acetylglucosamine-binding protein. 179, 324 Macroxyproteinase characteristics. 186. 493 Magnesium 28Mg2detection, 173, 548 properties, 173, 547 subcellular distribution, determination. 173, 553 in rapidly frozen cells, electron probe Xray microanalysis, 172, 203 transport in eukaryotic and prokaryotic cells, assay, 173, 546 Magnetic fields orientation of synthetic and native membranes, 172, 406 Magnetic flux constant. MS scans with, 193, 47 Maize leaves intercellular fractionation, 174, 543 protoplast isolation, 181. 153

Malaria Malaria circumsporozoite protein overlapping peptide studies of immunogens, 178, 627 T-cell antigenic sites, identification, 178, 623 induced amino acid transport systems in red cells, 173, 129 parasites, isolation with free-flow electrophoresis, 171, 521 m-Maleimidobenzoyl-N-hydroxysuccinimide ester in conjugation of B- and T-cell epitopes, 178, 665 Maleimidobiocytin derivatives, preparation, 184, 132 Maleimidohexanoic acid chloride in leukotriene B4 immunogen preparation, 187, 121 in leukotriene C4 immunogen preparation, 187, 118 3-(N-Maleimidopropionyl)bioc ytin biotinylation of /3-galactosidase, 184, 154 thiol-containing proteins, 184, 153 preparation, 184, 131 3-(N-Maleimidopropionyl)biocytin hydrazide preparation, 184, 137 3-(N-Maleimidopropionyl)biocytin Nhydroxysuccinimide ester preparation, 184, 135 L-Malic acid chloralide synthesis, 188, 387 L-Malic acid chloralide chloride synthesis, 188, 388 Malondialdehyde absorptiometry, 186, 423 assay as index of lipid peroxidation, 186, 421 with thiobarbituric acid, 186, 408 fluorometric assay, 186, 431 in food, HPLC assay, 186, 428 gas chromatographic assay, 186, 430 HPLC assay, 186, 413 in tissues, HPLC assay, 186, 428 in urine, HPLC assay, 186, 429 Malpighian tubules dissection, 192, 620 fluid transport, analysis and control, 192, 617

142 functions, 191, 168 ion channels, patch-clamp studies, 191, 181 isolation, 191, 170 lumen, perfusion, 192, 627 osmotic permeability measurements, 192, 630 saline for, 192, 619 secretion studies in vitro, 191, 170 solute transport, analysis and control, 192, 617 transepithelial potential measurement, 192, 629 tubule wall permeability analysis, 192, 623 in vitro studies, validation, 192, 625 Maltoheptaose in analysis of sputtering of alkyl esters of p-aminobenzoic acid, 193, 668 collision-induced dissociation spectra, 193, 695, 700 L-Malylchloralidyl-N-octanoylcysteamine synthesis, 188, 389 L-4-Malyl CoA synthesis, 188, 386 Malyl-CoA lyase assay in crude extracts of methylotrophs, 188, 364 Methylobacterium extorquens AMl assay, 188, 379 properties, 188, 383 purification from 188, 382 L-Malyl-N-octanoylcysteamine synthesis, 188, 390 Mammalian cells cotransformation by CaPO4-mediated cotransfection, 185, 563 direct DNA transfer into, 185, 495, 559 DNA introduction, 185, 527 heterologous genes expression experimental approach, 185, 508 overview, 185, 485 vectors for, 185, 487 selection and coamplification, 185, 537 lysates, preparation, 185, 579 permeabilization to macromolecules, 171, 857 proteins expressed in, synthesis, processing, and secretion, 185, 577

143 velocity sedimentation at unit gravity, 171, 501 Manduca sexta, see Tobacco hornworm Manganese in calibration of quin 2 fluorescence for assay of cytosolic free Ca 2+, 172, 246 -polyaminocarboxylate complexes, reactions with free radicals, 186, 117 in superoxide dismutase mimic prepared from MnO2 and desferrioxamine assay, 186, 244 valence, 186, 245 Manganese dioxide and desferrioxamine, derived superoxide dismutase mimic preparation, 186, 243 properties, 186, 244 Mannan endo- 1,6-/3-mannosidase purification from Bacillus circulans, 185, 461 Mannooligosaccharides O-linked, from chitinase, analysis, 194, 691 Mannoproteins associated N-linked oligosaccharides isolation, 185,457 NMR characterization, 185, 457 selective chemical degradation, 185,466 structural analysis with glycosidases, 185,459 yeast mnn mutant, isolation, 185,455 Mannose -N-acetylglucosamine-binding proteins, purification from mammalian sera, 179, 310 -L-fucose/N-acetylglucosamine-binding protein filter assay, 179, 323 isolation from rat peritoneal macrophages, 179, 324 3H-labeled, labeling of yeast glycoproteins. 194, 695 Mannose-6-phosphatase latency, in assessment of intactness of endoplasmic reticulum, 174, 59 Mannose-6-phosphate receptors relationship to insulin-like growth factor II receptor, 168, 323 a-Mannosidase Golgi apparatus marker, assay, 182, 219

c~- 1 , 6 - M a n n o s y l - g l y c o p r o t e i n nonspecific, purification from Oerskovia sp., 185, 464 yeast vacuole, assay, 194, 657 Mannosidases acting on yeast mannoproteins, general assay, 185,461 type i assay, 179, 464 properties, 179, 467 purification from mung bean microsomes. 179, 465 type IA assay, 179, 446 properties, 179, 449 purification from rat liver Golgi membranes. 179, 447 type 11 assay, 179, 468 properties, 179, 470 purification from mung bean microsomes, 179,469 ~- 1,3-Mannosyl-glycoprotein/3-1.2-Nacetylglucosaminyltransferase assay, 179, 363 properties. 179, 367 purification from rabbit liver, 179, 364 ~- 1,3-Mannosyl-glycoprotein/3-1,4-Nacetylglucosaminyltransferase assay, 179, 377, 405 chicken oviduct, properties, 179, 377 distribution in rat tissues, 179, 406 properties, 179, 405 c~-1,6-Mannosyl-glycoprotein/3-1.2-Nacet ylglucosaminyltransferase mung bean microsomal assay, 179, 472 properties, 179, 474 purification, 179, 473 rat liver assay, 179, 368 properties, 179, 373 purification, 179. 369 ~- 1,6-Mannos yl-glycoprotein/3-1,4-Nacetylglucosaminyltransferase assay, 179, 380 properties. 179, 381 c~-1,6-Mannosyl-glycoprotein/3-1,6-Nacet ylglucosaminyltransferase assay, 179, 378. 406 distribution in rat tissues, 179, 407 properties. 179, 379,407

fl- 1,4-Mannosyl-glycoprotein fl- 1,4-Mannosyl-glycoprotein/3-1,4-Nacetylglucosaminyltransferase assay, 179, 374,400 chicken oviduct, properties, 179, 375 distribution in rat tissues, 179, 404 properties, 179, 402 Mannosyl-glycoprotein endo-/3-N-acetylglucosaminidase applications, 179, 515 assay, 179, 506 properties on glycoprotein substrates, 179, 511 purification from F l a v o b a c t e r i u m m e n i n g o s e p t i c u m , 179, 506 Mannosyltransferase assay, 185, 469 solubilized, preparation, 185, 468 Markov model for molecular evolution, theory, mathematical formulation, and applications, 183. 571 Mass, see also Molecular weight amino acid residues in peptides, 193, 888 exact spectrometric measurements, utility, 193, 295 isotopes, values, 193, 869 molecular membrane components, determination by target analysis, 172, 410 proteins, determination by 252Cf plasma desorption MS, 193, 269 reference ions, 193, 870 reference standards, selection and applications, 193, 305 Mass analyzers electric sector, overview, 193, 38 magnetic sector, overview, 193, 40 quadrupole, overview, 193, 56 time-of-flight, overview, 193, 53 Mass spectra electron ionization, origin, 193, 6 and selectivity, differences, 193, 32 Mass spectrometry angle-resolved, collision-induced dissociation, 193, 250 array detectors, 193, 78 chemical derivatization for, 193, 314 chemical ionization -HPLC, glycosphingolipids, 193, 725 measurements, 2H introduction for, 193, 332

144 partially methylated alditol acetates, 193. 571 -supercritical fluid chromatography, glycosphingolipids, 193, 726 collision-induced dissociation high-energy oligosaccharides, 193, 689 overview, 193,240 low-energy methods, 193, 251 desorption glycosphingolipids, 193, 713 oligosaccharides coupled with hydrophobic chromophores, 193, 661 peracetylated and permethylated, 193, 647 direct measurement detectors, 193, 62 dithiothreitol-dithioerythritol mixture, 193, 878 DNA preparation and enzymatic hydrolysis for, 193, 782 electron-capture negative-ion chemical ionization lipid mediators, 187, 13 6-oxo-prostaglandin F~, 187, 20 electron ionization measurements, 2H introduction for, 193, 332 methylated/acetylated 1,5-anhydro-ohexitols, 193, 576 partially methylated alditol acetates, fragmentation rules, 193, 564 trimethylsilyl derivatives of nucleosides, 193, 825 electrospray ionization description 193, 412 interface with capillary separation schemes, 193, 430 negative ion mode, 193, 423 positive ion mode, 193, 419 proteins and Mr determination, 193, 421 energy-resolved, collision-induced dissociation, 193, 254 exact mass measurements, utility, 193, 295 fast atom bombardment, see Fast atom bombardment mass spectrometry Fourier transform, collisional activation in, 193, 259 glucuronides, 189, 142

145 glycerol, 193, 876 glycoconjugates, overview, 193, 539 glycosphingolipids ceramide composition, analysis, 193, 637 intact mixtures, 193, 623 permethylated mixtures, selected-ion curves, 193, 632 permethylated-reduced mixtures, selected-ion curves, 193, 627 -HPLC, mass reference standards, 193, 310 hybrid, collisional activation in, 193, 258 liquid secondary ion in amino acid sequencing, 193, 490 in analysis of Tyr phosphorylation in angiotensin by pp6W c, 193,485 analyte modification, 193, 210 artifacts, 193, 209 in assay of phosphate or sulfate in proteins and peptides, 193,487 and automated sequencing, for phosphoserine identification, 193, 492 derivatized oligosaccharides, 193, 668 in detection of phosphate or sulfate in proteins and peptides, 193,482 in estimation of peptide phosphorylated/dephosphorylated ratio, 193, 489 glycosphingolipids, 193, 723 ionization process, 193, 208 mass reference standards, 193, 311 matrix selection, 193, 201 spectra, 193, 875 measurements, 2H introduction for, 193, 334 phosphorylated and sulfated peptides and proteins, 193, 480 sputtering process, 193, 203 sulfated Tyr-containing peptides, 193, 486 -TLC, glycosphingolipids, 193, 725 tryptic peptides from glycogen synthase, 193, 483 magnetic, selected-ion monitoring, mechanics, 193, 99 measurements, 2H introduction for, 193, 329 methyl retinoate, 189, 104

Mass spectrometry molecular secondary ion, particle beam ionization methods, 193, 25 multiplier detectors, 193, 65 2-nitrobenzyl alcohol, 193, 879 2-nitrophenyl octyl ether, 193, 880 nucleic acid constituents, overview. 193. 771 nucleic acids, preparation of trimethylsilyl derivatives for, 193, 791 in peptide sequencing, 193, 407 plasma desorption 252Cf, proteins, 193, 263 particle beam ionization methods. 193. 24 sample preparation, 193, 432 peptides molecular weight determination. 193. 354 overview, 193. 351 problems requiring sequence information, 193, 359 prostaglandin H:, 187, 246 proteins enzymatic and chemical digestion It)r. 193, 361 mixture spectra, analysis, 193. 448 molecular weight analysis methodology, 193, 441 utility, 193, 354 overview, 193, 351 problems requiring sequence information, 193, 359 recombinant, 193, 523 quadrupole ion trap, collisional activation in, 193, 258 quadrupole, selected-ion monitoring, mechanics, 193, 101 resolution, single-ion curve window, and fragment specificity, 193. 641 retinol, 189, 101 RNA preparation and enzymatic hydrolysis for, 193, 782 selected-ion monitoring, overview, 193, 86 sensitivity enhancement, 193, 641 stable isotope dilution in assessment of vitamin A status, 189, 94 epoxy- and dihydroxyeicosatrienoic acids, 187, 175 9c 187, 51

Mass spectrometry sulfolane, 193, 881 tandem, see Tandem mass spectrometry thermospray nucleosides, 193, 807 spectra acquired in D.,O, 193, 812 1-thioglycerol, 193, 877 time-of-flight, selected-ion monitoring, mechanics, 193, 102 -TLC, assessment, 193, 642 uv laser desorption/ionization, matrix assisted, peptides and proteins, 193, 280 Mast cells bone marrow-derived activated, lipid metabolite production, 187, 517 cell culture, 187, 515 exocytosis of lipid mediators, 187, 516 phosphatidylinositol turnover, 187, 517 stimulated, 5-1ipoxygenase activation and exocytosis, differentiation, 187, 518 metabolism of albumin-bound leukotriene A 4 to C4, 187, 575 -neutrophil interactions: leukotriene A4 release and metabolism, 187, 567 sulfidopeptide leukotrienes, assay, 187, 97 Mating yeast and complementation, 194, 18 nonmating mutants, 194, 91 reaction assay, 194, 77 switching of types, applications bilateral mating defect testing, 194, 138 facilitation of complementation tests, 194, 137 isogenic strain construction, 194, 136 methodology, 194, 132 polyploid strain construction, 194, 138 Matrices affinity, see Affinity matrices for gel filtration of proteins, 182, 320 solute-absorbing, in protein concentration and solute removal, 182, 81 Matrix patterns in determination of DNA regulatory site consensus patterns, 183, 213

146 Maximum entropy method Burg, for NMR spectral estimates, 176, 218 and other non-Fourier transform methods, comparison, 176, 228 Maximum entropy reconstruction for NMR spectral estimates, 176, 225 and other non-Fourier transform methods, comparison, 176, 228 Maximum likelihood estimation in analysis of data from in situ hybridization studies, 168, 837 reconstruction of phylogenetic trees for nucleotide sequences, 183, 584 MCV VERSUS TIME program calculation of permeability coefficient to water, 172, 275 Megakaryocytes human marrow, detection of histidinerich glycoprotein, 169, 273 interaction with subendothelial extracellular matrix, 169, 87 Meiosis chromosome segregation in, analysis with yeast artificial chromosomes, 194, 765 in Saccharomyces cerevisiae entry-regulating genes and mutations, identification, 194, 104 monitoring, 194, 94 mutational analysis, 194, 99 Melting absorbance curves for RNA, determination, 180, 304 RNA, related behavior analysis, 180, 322 prediction, 180, 319 thermodynamic parameters, 180, 315 Membrane potential ascites tumor cells, assay with carbocyanine dyes, 172, 95 in cells, optical measurement, 172, 102 with defined magnitude, creation in intestinal epithelial cells, 192, 337 in determination of paracellular shunt conductance in epithelia, 191, 4 in epithelium, measurement, 171,772 EPR measurement, 172, 331 gall bladder epithelium, measurement, 192, 657

147 giant algal cells control, 174, 413 measurement, 174, 406 hepatocyte plasma membrane, effect on alanine transport, 174, 35 intestinal epithelial cells, measurement, 192, 338 intestinal secretory cells, measurement, 192, 311 measurement, 172, 64 micro- and macroelectrodes measuring, preparation and use, 172, 136 mitochondrial, in heart during ischemia and reperfusion, 186, 723 muscle fibers, in assay of Na+-K * pump activity, 173, 710 in Nitella, measurement, 174, 480 organelles, optical measurement, 172, 102 pancreatic 13 cells, measurement, 192, 235 plant, measurement with pH electrode, 174, 298 relationship to short circuit current, 191, 12 red blood cells, measurement, 173, 94 in Rhodotorula, measurement, 174, 633 vesicles, optical measurement 172, 102 yeast, determination with carbocyanines, 174, 615 lipophilic anions, 174, 616 microelectrodes, 174, 611,650 quaternary phosphonium compounds, 174, 611 Membrane proteins amphipathic, bilayer spanning domains, prediction, 172, 439 bound detergent, determination, 172, 526 carrier, isolation, 171, 17 channels, mediated ion movement, kinetics, 171, 62 characterization, 184, 629 concentration, effects on quaternary structure studies, 172, 612 crosslinking principles, 172, 611 reagents, 172, 586 sterically enforced reactions, 172, 605 topographical studies, 172, 608 -detergent complexes, size and shape, hydrodynamic studies, 172, 696

Membranes detergent removal, 182, 277 functionally active, renaturation, 182, 516 functional modulation by surface potential, 171,387 genetic analysis, 182, 517 hydrophobic bilayer spanning domains, prediction. 172,439 folding pattern, 172, 440 immunological analysis, 182, 517 integral fractionation, 182, 508 and membrane-embedded domains, photochemical labeling, 172, 630 reconstitution. 182, 516 solubilization, 182, 505 spontaneous insertion into preformed unilamellar vesicles, 171, 253 localization in plasma membrane. 184. 637 membrane skeleton, preparation from red cells, 173, 380 Triton shells, 173, 389 microsomal, solubilization, 182, 232 mitochondrial, direct interaction with triiodothyronine, 174, 128 molecular weight determination, 172, 514 native, solubilization, 182, 253 peripheral, purification, 182, 504 in-plane ordering, 172, 409 purification, 182, 499; 184, 629 resistance to proteases, 191, 852 SDS-induced dissociation, 191, 850 sample concentration and analysis, 182, 518 secondary structure and aggregation state, determination by circular dichroism, 172, 575 solubilization by nonionic surfactants, 172, 519 solubilized, extinction coefficient at 280 nm, determination, 172, 524 target analysis, literature data tabulation, 172,429 transmembrane reconstitution, 171, 265 Membranes acinar cell, preparation, 192, 29 amino acid transport, heterogeneity of

Membranes systems for, recognition and resolution, 173, 576 anion transporter orientation, 173, 423 apical epithelial, ionic permeation measurements, 171, 680 pathways, 171, 711 gall bladder, ion and water transport, patch clamp studies, 192, 676 goblet cell isolation from tobacco hornworm midgut, 192, 609 purification of vacuolar-type ATPase, 192, 612 turtle colon, ion channels, analysis, 192, 737 vesicles, isolation from renal epithelial cell lines, 191, 501 apolar phase, photochemical labeling, 172, 628 basement, gel threads, in NMR studies of intracellular metabolism, 177, 440 basolateral contamination with brush border membranes, 172, 352 effects of different vesicle preparations, 192, 450 epithelial, ionic permeation measurements, 171, 680 pathways, 171, 713 gall bladder, ion and water transport, patch clamp studies, 192, 678 highly purified, four-dimensional isolation, 192, 350 intestinal epithelial cell, Ca2+ transport, 192, 448 partially purified, isolation, 192, 351 properties, 192, 453 turtle colon, transporter identification and characterization, 192, 739 bilirubin bound to, antioxidant activity, 186, 306 binding of [~25]iodo-ovineprolactin, potency of inhibiting anti-Id antibodies, 178, 316 biological, surface potential determination, 171, 364 effects on protein function, 171, 387 lipid coumarin dye as probe, 171, 376

148 bound prolactin receptor fractions, preparation, 178, 313 brush border contamination with basolateral membranes, 172, 349 divalent cation-free, isolation, 192, 353 extraction, 192, 440 intestinal, perturbation and photoaffinity labeling by azides, 191, 767 isolated, in analysis of stimulussecretion coupling, 191, 675 isolation by divalent cation precipitation, 192, 353 for glucose transporter studies, 192, 439 for NaCI transport studies, 192, 392 purification from renal cortex, 191, 451 renal, perturbation and photoaffinity labeling by azides, 191, 767 vesicles in analysis of transport systems, evaluation, 192, 415 functional, preparation, 192, 410 transport kinetics, 192, 422 transport measurements, 192, 413 cellular effects of photoaffinity-labeling analogs of phlorizin and phloretin, 191, 755 isolated incorporation of labeled phosphoinositides, 168, 341 phosphoinositide hydrolysis, assay, 168, 338 preparation from B cells, 168, 340 leaks, effects on impedance analysis, 171, 657 ApH determination, 174, 131 cerebral, opioid receptor binding studies, 178, 251,256 for chemotaxis assays, cell counting on, 169, 241 colonic cell, electrophysiology, 192, 482 components, radiation inactivation and molecular mass determination, 172, 410 dialysis, types and applications, 168, 188 dynamics, EPR measurement, 172, 331

149 electrostatic potentials adjacent to, measurement, 171, 342 envelope, Sendai virus mediated fusion for carrier and receptor implantation into cells, 171, 829 reconstituted fusogenic activity, estimation, 171, 839 preparation, detergent selection for, 171, 842 reconstitution, 171, 835 enterocyte, isolation, 192, 341 epithelial apical and basolateral, ionic permeation measurement, 171, 680 pathways, 171, 711 gall bladder, ion and water transport, patch clamp studies, 192, 676 intestine, Ca `,+ transport, 192, 448 ionic selectivity, 171, 727 resistance properties, determination, 171, 719 erythrocyte anion-exchange protein isolation, reconstitution, and transmembrahe orientation, 173, 410 bulk D20 diffusion across, 173, 199 chemical equilibrium, relationship to cell conformation, 173, 12 cytoskeletal protein removal, 174, 41 failure mechanisms, 173, 29 ghosts, s e e Erythrocytes, ghosts human, cytoplasmic surface, protein associations with band 3,173, 513 inside-out vesicles Na*,K+-pump stoichiometry and coupling, 173, 377 preparation and properties, 173, 368 intact, macromolecular crosslinking, 184, 158 lipid core, photochemical labeling, 172, 659 material properties, 173, 8 mechanical properties, direct measurement, 173, 17 nucleoside transport, 173, 250 osmotic water flow across, 173, 204 preparation, 174, 40

Membranes reassociation of protein ligands with, 173, 516 skeleton proteins, preparation, 173. 38O water diffusion across, 173, 192 excised patches, single-channel currents recorded in, 171, 665 fractions, preparation, 182, 225 gall bladder cell ion selectivities, 192, 659 voltage and resistance measurements, 192, 657 gastric cell, permeability, 171,459 gastric gland, preparation, 192, 155 Golgi, rat liver, purification of mannosidase IA, 179, 446 hepatic, crude, for antibody mapping of sodium channels, preparation, 178, 725 hepatocyte, vesicles bile acid transport mechanisms, 192. 517 isolation, 192, 517 H ÷,K*-ATPase-containing isolated gastric, purification, 191, 734 isolation from gastric oxyntic cell, 192, 151 pH gradient formation, assay, 192, 161 hydrophobic core, chromanol incorporation, 186, 362 hydrophobic photolabeling applications, 172, 658 experimental aspects, 172, 653 intestinal epithelial cell, Ca 2+ transport, 192, 448 intestinal secretory cell electric current effect of intracellular Ca2% 192, 316 measurement, 192, 311 K + conductance characterization, 192, 309 pharmacological properties, 192, 316 intestinal, vesicles CI transport, 192, 405 electrophysiology, 192, 404 NaCI transport pathways, 192, 389 Na + transport, 192, 399 ion movement mediated by carriers and channels, kinetics. 171, 62

Membranes ion transport, assay on individual protoplasts, 174, 322 labeling by monocarboxylate transport inhibitors, 173, 326 leaflets, [~4C]phosphatidylcholine equilibration between, 173, 227 lipid core, labeling reagent requirements, 172, 633 liposomal, transfer of retinol from retinol-binding protein across, 189, 394 microsomal hepatic, preparation, 168, 311 isolation and solubilization, 182, 231 tocopherol and tocopheryl ester incorporation and hydrolysis, 186, 355 yeast, protein translocation in vitro, assay, 194, 675 mitochondrial inner Ca 2-~ transport studies, 174, 107 proton leakage, measurement and relation to protonmotive force, 174, 85 tocopherol and tocopheryl ester incorporation and hydrolysis, 186, 355 nervous system tissue, neurotensin binding, assay, 168, 464 neutrophil, phospholipase C-catalyzed inositol trisphosphate production, assay, 187, 232 nutrient, HCO3- and CI- transport, mechanistic analysis, 171, 622 order and viscosity, analysis by timeresolved fluorescence depolarization, 172, 462 orientation for time-averaged and resolved structure determinations, 172, 399 ouabain-sensitive hyperpolarization, in assay of Na+-K + pump activity, 173, 712 parietal cell, intracellular ion activities and membrane transport, assay, 192, 38 permeability, unstirred layers determination, validity, 173, 166 perturbation by azides, 191, 767 phospholipid artificial, surface potential determination, 171, 342

150 coated cytoskeleton characterization, 173, 395 preparation by erythrocyte reconstitution, 173, 392 interactions with retinoids ESR studies, 189, 383 optical spectroscopic studies, 189, 373 pituitary, purification of gonadotropinreleasing hormone receptors, 184, 288 planar electrical conductance, 171, 361 lipid-protein, formation, 171, 225 plasma, s e e Plasma membranes platelet binding of thromboxane Ajprostaglandin H2 receptor antagonists, 187, 412 crude preparations, 187, 399 rabbit, ginkgolide interactions with PAF binding sites, 187, 435, 438 solubilized, preparation, 187, 399 surface, interaction with extracellular Ca 2+, 169, 355 polypeptide sites, ion interactions at, NMR studies, 171, 286 populations, markers, 172, 29 porcine aortic smooth muscle, preparation, 187, 400 preparation for purification of associated proteins, 182, 502 proximal tubule efflux coefficients, 191, 212 permeability estimates, 191, 210 pulmonary, preparation, 187, 416 rat tissue, associated bile salt-independent retinyl ester hydrolases assay, 189, 460 properties, 189, 461 renal preparation, 191,574 solute transport, assay, 191, 430 tubule cells amplification, 191, 269 recycling, 191, 271 receptor interaction with anti-idiotypic antibodies, 178, 179 resealable, maximally depleted of cytosolic components, preparation, 173, 346

15 l retinal, retinoid transfer, 189, 411 rod outer segment, retinoid exchange with lipid vesicles, 189, 402 rotational diffusion in, fluorescence and phosphorescence measurements. 172, 472 salivary gland, preparation, 192, 29 structure permeability-based analysis, 174, 265 relationship to channel function, assays, 171, 240 subcellular, isolation, 172, 19 surface charge, effects in analysis of ionic permeation mechanisms, 171, 700 translational diffusion in, fluorescence and phosphorescence measurements, 172, 495 transport processes carrier-linked, kinetic criteria, 171, 14 experimental paradigms, 171, 24 kinetics, 171, 23 machinery, overview, 171, 3 models, 171, 33 regulation by endocytotic removal and exocytotic insertion of transporters, 172, 49 system identification, isolation, and characterization, 171, 12 vacuolar, s e e Tonoplast vesicles, s e e Vesicles water channels, identification, 191, 561 yeast structures, visualization, 194, 607 surface potential effects on transport parameters, 174, 581 Menadione induced H202 formation, assay in v i t r o , 186, 625 Mercaptan specific antioxidant protein assay, 186, 481 properties, 186, 483 purification from yeast, 186, 481 Mercaptoethylamine radioprotective effects in mice, 186, 686 5-Mercaptouridine 5'-triphosphate synthesis, 180. 399 Mercury effect on retinoid binding by parasite retinoid binding protein, 189, 349

Metabolism Meromyosin heavy, biotinylated site, avidin attachment, 184, 413 Mesangial cells glomerular cultured, eicosanoid biochemistry, 187, 544 isolation and culture, 191, 146 primary cell culture, 187, 547 structure and function. 187, 544 Mesentery feline, leukocyte adherence to microvasculature, intravital assessment, 186. 733 Mesophyll cells wheat leaf, isolation of deoxyribonucleoprotein, 170, 441 Metabolism alanine, regulation, role of alanine transport, 174, 33 arachidonate by ocular cytochrome P450, 187, 372 in parietal cells. 187, 505 in rabbit renomedullary cells, 187, 365 arachidonic acid in bone marrow-derived mast cells, 187, 514 cultured cells, 187, 535 endothelial cells, 187, 531 human platelets, 187, 589 platelet and lymphocyte preparations. 187, 582 eicosanoids in isolated perfused rat lung, 187, 608 related enzymes, nomenclature, 187, 3 furosemide-related diuretics, 191, 789 inositol lipids in endocrine cells, analysis by transient permeabilization, 168, 218 intracellular, NMR analysis agarose gel thread method, 177, 437 basement membrane gel method, 177, 440 in perfused cells, 177, 437 in perfused organs, 177,441 phospholipids, 177, 449 in isolated renal tubule segments, 191, 325 leukotriene A4 in neutrophil-mast cell interactions, 187, 567

Metabolism neutrophil-vascular cell interactions, 187, 565 leukotrienes by isolated rat hepatocytes, 187, 277 oxidative, arachidonic acid in bone marrow-derived mast cells, 187, 514 phosphatidylinositol-derived arachidonic acid, 191, 676 quinones, effects on cellular glutathione and protein thiol status, 186, 627 retinal, in retinoic acid formation in epidermal cells, 189, 530 retinoic acid by intact cells and cell extracts, 189, 525 retinoids related alcohol dehydrogenase isoenzymes assay, 189, 437 purification, 189, 438 starch gel electrophoresis, 189, 438 in retinal pigment epithelial cells in vitro, 190, 16 role of liver cells, 190, 60 retinol by intact cells and cell extracts, 189, 525 in microsomes associated effects, 189, 482 products, HPLC, 189, 485 in retinoic acid formation in epidermal cells, 189, 530 transition metals, overview, 186, 29 versus transport, assay in cultured cells, 173, 714 Metal ions, see also specific ions binding to phosphoproteins, 3~p NMR study, 177, 279 proteins, quadrupolar NMR study, 177, 325 catalytic, role in human disease, 186, 1 role in .OH production by Of-generating and other systems, 186, 17 Metalloproteins paramagnetic JH NMR, 177, 246 NOE studies, 177, 260 Metalloth'ionein copper-induced promoter construction, 185, 321

152 in induced gene expression in yeast, 185, 319 induction conditions, 185, 324 internal and 5' deletions, 185, 323 encoding gene, isolation from yeast, 185, 321 promoters for expression of cDNA genes in mammalian cells, 185, 498 Metals alkali, see Alkali methals catalytic, detection in simple buffers, 186, 125 complexes, application in neuroendocrine studies, 168, 710 higher oxidation states, superoxidedependent formation, 186, 27 transition, see Transition metals Metapyrocatechase, see Catechol 2,3dioxygenase Methane monooxygenase Methylosinus trichosporium OB3b assay, 188, 191 component protein concentrations, optimization for assays, 188, 192 properties, 188, 200 purification, 188, 197 soluble, from Methylococcus capsulatus Bath assay, 188, 181 component proteins, assay and purification, 188, 183 properties, 188, 186 purification, 188, 183 Methanol metabolizing enzymes, cytochemical staining in Hansenula polymorpha, 188, 411 peroxidation, in assay of drug-dependent H202 generation, 186, 622 Methanol:cytochrome c oxidoreductase assay, 188, 212 Methanol dehydrogenase Bacillus CI assay, 188, 223 properties, 188, 225 purification, 188, 224 Hyphomicrobium X

assay, 188, 203 properties, 188, 206 purification, 188, 204

153 Methylobacterium extorquens AMI

assay, 188, 210 properties, 188,215 purification, 188, 213 modifier protein assay, 188, 217 properties, 188, 221 purification from methylotrophs, 188, 218 separation from cytochromes CHand CL, 188, 304 in stopped-flow spectrophotometric assay of cytochromes cH and Ce, 188, 304 Methanolysis permethylated oligosaccharides, 193, 651 Methemoglobin absorption spectra, 186, 268 Methidiumpropyl-EDTA source and storage, 170, 362 Methidiumpropyl-EDTA.iron(II) digestion of nuclei, 170, 363 preparation, 170, 363 Methionine effect on sinusoidal transport of glutathione, 173, 528 3H-labeled, in pulse labeling of prerRNAs, 180, 99 in proteins, spin labeling, 177, 116 spin system identification, 177, 139 symport of Na + with, demonstration, 173,774 Methionine aminopeptidase isolation from prokaryotes, 185,407 Methionine-tRNA ligase bound reactants and products, equilibrium constant determination, 177, 372 Methionyl-tRNA synthetase, see Methionine-tRNA ligase Methotrexate resistance, in selection for dihydrofolate reductase gene amplification, 185, 544 Methoxime derivatives, formation for MS analysis, 193, 327 3-Methoxyglucose equilibrium exchange in red cell, 191, 771

Methyl epoxyeicosatrienoic acids Nr-Methyladenosine antibodies in affinity chromatography of snRNPs U2 and U4/U6, 181, 244 purification, 181, 236 in purification of snRNP particles, 181, 232 Methylamine dehydrogenase blue copper proteins as electron acceptors for, assay, 188, 285 Methylobacillus flagellatum

assay, 188, 247 properties, 188, 249 purification, 188, 248 methylotrophic bacterial assay, 188, 242 properties, 188, 245 purification, 188, 242 subunit resolution and reconstruction. 188, 245 Thiobacillus versutus

assay, 188, 236 properties, 188, 239 purification, 188, 236 Methylamine oxidase, see Amine oxidase (copper-containing) N-Methylaniline demethylation by cumyl hydroperoxide, assay, 186, 274 Methyl arachidonate epoxidation, 187. 361 Methylation in analysis of O-linked oligosaccharides, 179.22 carbohydrates, 193, 556 carboxyl, glutamate and aspartate residues, 182, 632 pre-rRNA in isolated nuclei, 180, 107 5-Methylcytosine in DNA, isotope dilution GC/MS, 193, 857 labeled, synthesis, 193, 859 localization in HI-containing nucleosomes, 170, 248 Methyl 6-deoxy-6-amino-B-D-galactopyranoside formation from methyl/~-D-galactopyranoside, 179, 258 Methyl epoxyeicosatrienoic acids saponification, 187. 363

Methyl esters Methyl esters formation by lipid peroxide transesterification, 186, 390 mass spectrometry, 193, 325, 328 Methyl fl-o-galactopyranoside conversion to 6-amino derivative for attachment of functional groups, 179, 258 a-Methylglucoside accumulation by intestinal epithelial cells, 192, 329 Methylglycoside collision-induced dissociation spectra, 193, 697 Methyl linoleate hydroperoxides 13C NMR assay, 186, 380 and dienols, HPLC, 186, 381 2-Methyl-6-(p-methoxyphenyl)-3,7-dihydroimidazo[1,2-a]pyrazin-3-one chemiluminescence, in assay for superoxide dismutase, 186, 227 N-Methyl-N'-nitro-N-nitrosoguanidine induction of mutagenesis in yeast, 194, 277 N-Methyl-N-nitrosourea induction of breast cancer, 190, 399 Methylobacillus flagellatum purification of glucose-6-phosphate dehydrogenase, 188, 335 methylamine dehydrogenase, 188, 248 6-phosphogluconate dehydrogenase, 188, 338 Methylobacterium M. extorquens AMI, purification of hydroxypyruvate reductase, 188, 374 malyl-CoA lyase, 188, 382 methanol dehydrogenase, 188, 213 soluble cytochromes c, 188, 301 M. organophilum XX, purification of serine hydroxymethyltransferases, 188, 368 Methylococcus capsulatus Bath purification of soluble methane monooxygenase, 188, 183 Methylomonas A4 purification of soluble cytochromes c, 188, 291 Methylophilus methylotrophus purification of

154 electron-transfer flavoproteins, 188, 309 methanol dehydrogenase M protein, 188, 218 Methylosinus trichosporium OB3b purifcation of formate dehydrogenase, 188, 332 methane monooxygenase, 188, 197 Methyl red induced H202 formation, assay in vitro, 186, 625 Methyl retinoate mass spectrometry, 189, 104 Methyl sulfoxide carbanion preparation, 193, 556 Metmyoglobin ZH NMR spectrum, 177, 249 Micelles mixed aggregated, extraneous protein removal, 191, 601 /3-carotene incorporation, 189, 418 solutions, preparation, 189, 419 Microautoradiography in tracing assimilates from mesophyll to phloem, 174, 294 Microbeads cationic silica, in isolation of Rhodotorula plasma membrane fractions, 174, 648 Microcapsules preparation from human erythrocytes and application in transport studies, 171, 217 Microcentrifugation in silicone oil, in assay of Mg2+ transport in suspension culture, 173, 551 Micrococcal nuclease in assembly of chromatin with oocyte extracts, 170, 611 in analysis of RNA-protecting factor interactions, 180,465 digestion of nuclei, 170, 329 inactivation of histone-specific RNA processing, 181, 82 in preparation of chromatin, 170, 127 in yeast mRNA titration, 194, 539 Microdialysis application in vivo, 168, 176

155 in assay of hypothalamic releasing factors, 168, 168 neuroendocrine peptides, 168, 166 in neuroendocrinology in vivo experiments, 168, 195 principles, 168, 186 probes design and attached membranes, 168, 186 in vitro recoveries, 168, 191 Microdissection adult pancreas, 192, 189 brain tissue, 168,400 nephron segments, 191, 308 renal tubule segments, 191, 226, 328 Microelectrodes for analysis of ion and water transport in gall bladder epithelium, 192, 651 impalement of pancreatic fl cell, 192, 242 ion-exchanger, pH measurements, 174, 336 ion-selective in analysis of cell composition, 171, 769 preparation, 192, 664 for measurement of intracellular potential and resistance, 192, 86 ion activities, 172, 136 ionic permeation properties, 171, 685 pancreatic B-cell membrane potentials, 192, 235 pH, 174, 331 plasma membrane potentials, 171, 772 transmembrane potentials, 172, 136 yeast membrane potentials, 174, 611 Microfluorometry flow, pituitary CRF-target cells, 168, 34 Fura-2, 192, 60 Microinfusion in renal research, 191, 81 Microinjection antibodies into amphibian oocytes, 170, 241 somatic cells, 170, 245 DNA into mammalian cells, 185,535 erythroid band 3 protein mRNA into Xenopus oocytes, 173, 456

Micropuncture hepatocyte couplets and triplets, 192, 511 mRNA into giant MDCK cells, 191,443 nonpermeable compounds into cells, 191, 704 photoproteins into cells, 172, 171 in renal research, 191, 81 Micromanipulators for yeast asci, 194, 21 Microneedles for separation of ascospores, zygotes, and vegetative cells, 194, 28 Microorganisms partitioning in aqueous polymer twophase systems, 171,540 Microperfusion in assessment of single-nephron glomerular filtration, 191, 126 capillary, kidney in situ, 191, 105 capillary and lumenal, kidney in situ, 191, 104 distal tubules from reptile in vitro, 191, 220 double, kidney in situ, 191, 98 lumenal, stationary or continuous, kidney in situ, 191, 102 Malpighian tubules in vitro, 191, 174 proximal tubules from reptile in vitro, 191, 203 in renal research, overview, 191, 82 renal tubules from amphibians, 191, 197 fish, 191, 192 Micropipet glass, siliconization and protein coating, 191, 344 Microplate assay kinetic, superoxide production by phagocytic cells, 186, 567 Micropressure techniques in electrophysiological assay of opioid peptides in vivo, 168, 108 Microprojectiles in DNA transfer into mammalian cells. 185, 536 Micropuncture in assessment of single-nephron glomerular filtration, 191, 123 hepatocyte couplets, 192, 511 in renal research, 191.72

Microscopes Microscopes in yeast genetic analysis, 194, 28 Microscopy, see also specific techniques in analysis of volume regulation in epithelia, 171, 751 digital in imaging of nuclear DNA movement during yeast mitosis, 194, 750 in quantitation of fluorescence, 168, 276 in situ hybridization histochemistry, 168, 811,814 intravital, in assessment of leukocyte adherence and extravasation, 186, 733 isolated tubules, in measurement of volume and shape changes, 191, 371 liver stellate cells, 190, 70 phase, platelet counting with, 169, 165 renal epithelia in situ, 191, 273 in in situ hybridization histochemistry with markers of neuronal connectivity, 168, 789 visualization of renal epithelial cell fusion, 192, 721 yeast, 194, 572 Microsomes active in protein translocation, preparation, 194, 680 detection of chemiluminescence, 186, 597 hepatic gradients, in vesicle immunoadsorption, 191, 839 isolation from rat, 187, 386 NAD÷-dependent retinol dehydrogenase, detection and characterization, 189, 520 preparation, 187, 218 production of epoxy- and dihydroxyeicosatrienoic acids, assay, 187, 182 superoxide radical generation, assay, 186, 588 intestinal, isolation from rat, 189, 453 lipid peroxidation, inhibition by 13-cisretinoic acid, 190, 281 membranes isolation and solubilization, 182, 231 yeast, protein translocation in vitro, 194, 675

156 and mitochondria, differential rate sedimentation, 172, 23 mung bean, purification of N-acetylglucosaminyltransferase II, 179, 473 glucosidase I, 179, 457 glucosidase II, 179, 462 mannosidase I, 179, 465 mannosidase II, 179, 469 ocular, preparation, 187, 374 ovine seminal vesicle preparation, 187, 247 purification of prostaglandin H2, 187, 248 preparation, 179, 423; 189, 483 pulmonary guinea pig, purification of leukotriene C4 synthase, 187, 335 preparation and solubilization, 187, 254 retinal pigment epithelial, acylCoA:retinol and lecithin:retinol acyltransferases, assays, 190, 156 retinol metabolism associated effects, 189, 482 HPLC analysis, 189, 485 sulfation of proteochondroitin, chondroitin, and chondroitin oligosaccharides, 179, 422 Microspheres fluorescent biotinylated coating with streptavidin, 184, 355 -streptavidin, coating with biotinylated probe, 184, 355 biotinylation, 184, 354 in detection of cell surface components, 184, 353 Microsurgery on Acetabularia, 174, 491 in functional analysis of hypothalamohypophyseal axis, 168, 234 Microtiter assay methanol dehydrogenase, 188, 212 Microtiter plates in serological detection of anti-idiotypic antibodies, 178, 84 Microtiter wells cell adhesion to immobilized carbohydrates, 179, 547

157 Microvasculature injury, role of leukocytes, 186, 736 mesenteric, leukocyte adherence, intravital assessment, 186, 733 Microwells plastic, adsorption of glycosphingolipids, 179, 546 Midgut isolation of goblet cell apical membrane, 192, 609 lepidopteran, isolation, voltage clamping, and flux measurements, 192, 599 Milk bovine, purification of exocellular superoxide dismutase, 186, 249 isolation and identification of neuroendocrine peptides, 168, 494 Miller chromatin-spreading method in electron microscopy of ribonucleoprotein complexes on nascent RNA. 180. 481 Mimicry biological anti-idiotypic antibodies as vaccines in infectious diseases, 178, 7 and molecular mimicry, distinction, 178, 30 molecular, s e e Molecular mimicry network connectivity-based concept, 178, 33 network versus molecular, 178, 32 structural, reovirus type 3 hemagglutinin by anti-ld antibody to viral receptor, 178, 333 Mineralocorticoid receptors cytoplasmic, assay, 191, 529 kinetics and steroid specificity, 191, 521 nuclear binding assays, 191, 532 Minichromosomes simian virus 40 electron microscopy, 170, 166 preparation, 170, 14 for electron microscopy, 170, 166 in hypotonic conditions, 170, 19 in NaCl and EDTA, 170, 17 in NaCl and MgC12, 170, 19 purified, characterization, 170, 20 radiolabeling in uiuo, 170, 16

Mitochondria subpopulations characterization, 170, 24 visualization, 170, 177 Minilobules pancreatic, preparation and incubation, 191, 686 Mitochondria ADP/ATP exchange, characterization, 171, 12 calcium transport, analysis, 174, 68 cardiac membrane potential during ischemia and reperfusion, 186, 723 superoxide radical generation, assay, 186. 588 DNA, sequence branching: analysis of human evolution. 183. 563 effects of ~x-adrenergic agonists. 174, 119 glucagon, 174, 123 triiodothyronine, 174, 127 vasopressin, 174, 119 gene evolution, quantitative estimates: effect of base composition, 183, 578 H+-ATPase coupling rules, 171, 158 intrinsic uncoupling, nonequilibrium thermodynamics, 171,437 hepatic, rat, purification of phosphate transporter, 173. 732 inner membrane Ca 2+ transport, analysis. 174, 107 proton leakage, measurement and relation to protonmotive force, 174, 85 intact, enzymes in, Ca2+-sensitive properties, 174, 98 intrasynaptosomal, Ca 2+ levels, assay, 174, 7 isolation from mammalian cell lines, 182, 208 marker enzyme cytochrome-c oxidase, assay, 182, 214 matrix, entrapment of fluorescent Ca 2" indicators, 174, 115 membrane surface potential, determination, 171, 364 and microsomes, differential rate sedimentation, 172, 23 nonrespiring, proton permeability by swelling, 174. 86

Mitochondria

158 k

preparation for studies of Ca2+-sensitive intramitochondrial enzymes, 174, 97 renal cell, density and transport rate, 191, 272 staining in cells of isolated nephron segments, 191, 258 transmembrane electrical potential difference, determination, 172, 69 yeast DNA behavior in crosses, 194, 154 introns in, description, 181, 540 isolation and analysis, 194, 158 rho ÷, replacement of wild-type by mutant gene, 194, 164 enzyme marker assay, 194, 658 function and assembly, 194, 627 gene analysis and manipulation, 194, 149 gene expression, analysis, 194, 156 gene transformation and replacement, 194, 162 immunofluorescent detection, 194, 602 isolated, functional assays, 194, 634 isolation, 194, 631 mutations isolation, 194, 150 sequence analysis, 194, 160 preparation, 182, 173 protein import, 194, 629, 637 reverse genetics, 181, 551 RNA isolation and analysis, 194, 158 splicing, genetic analysis, 181, 539 Mitogenesis EGF-dependent, effect of ganglioside GM3, 179, 525 Mitosis cells at various stages, separation, 171, 496 in yeast, nuclear DNA movement during, digital imaging microscopy, 194, 750 Mixers dynamic and static, in HPLC of proteins, 182, 398 MNNG, see N-Methyl-N'-nitro-N-nitrosoguanidine Modification interference analysis reactions with R N A substrates, 180, 369

yeast pre-mRNA sequence contribution to splicing efficiency, 181, 143 Molecular dynamics in determination of molecular structure from NMR data, 177, 204 Molecular mimicry idiotope topography in, 178, 27 mimicry of contact residues, 178, 28 versus network mimicry, 178, 32 and receptors, relationship, 178, 21 relationship to biological mimicry, 178, 30 role in autoimmunity, 178, 19 schistosomal glycanic epitope by antiidiotypic antibodies, 178, 392 Molecular recognition theory in generation of anti-idiotypic antibodies of predefined specificity, 178, 64 Molecular size parasite retinoid-binding protein, profiles, 189, 349 Molecular structure conformational antigens, detection of changes, 178, 761 cellular retinol-binding protein, 189, 513 chemical shift dependence on, 176, 368 distance geometric analysis, 177, 159 epitopes, size exclusion HPLC analysis, 178, 120 Escherichia coli EtrT°-DNA complex, changes, 185, 40 proteins and mutants, comparison, 177, 74, 282 retinoic acid analogs, 189, 37 determination from NMR data by molecular dynamics simulation, 177, 204 quaternary analysis: effects of protein concentration, 172, 612 proteins, oxidative modification, 186, 491 secondary dependent gap penalties, in sequence alignment, 183, 408 hydropathy scales, comparison, 183, 454

159 membrane proteins, determination by circular dichroism, 172, 575 native ribonucleoproteins, probing, 180, 212 predictions, comparison, 183, 450 proteins /3 turn incidence, in prediction of B-cell determinants, 178, 591 NMR determination, 177, 223 oxidative modification, 186, 491 prediction, 182, 765,771 R-loops, interpretation, 180, 254 RNA computer modeling and display, 183, 318 computer prediction, 180, 262 enzymatic probing, 180, 192 optimal and suboptimal, prediction, 183, 281 phylogenetic analysis, 180, 227 pseudoknots, 180, 289 small nuclear, native, probing, 180, 212 -ssDNA heteroduplexes, interpretation, 180, 259 around viroid RNA self-cleavage sites, 181, 603 around virusoid RNA self-cleavage sites, 181, 593 yeast leader region, effect on translation initiation, 185, 368 tertiary proteins, oxidative modification, 186, 491 RNA computer modeling and display, 183, 318 enzymatic probing, 180, 192 Molecular weight, s e e a l s o Mass histone subunits, 170, 469 mass spectrometric determination, 193, 476 membrane protein polypeptide constituents, 172, 520 membrane proteins, 172, 514 peptides, 193, 354 proteins determination by gel electrophoresis, 182, 439

Monoclonal antibodies mass spectrometric determination electrospray ionization technique, 193, 421 overview, 193,441 utility, 193, 354 screening for gel filtration, 182, 322 synthetic oligonucleotides, 193, 778 Monactin ion carrier in planar bilayers, relaxation and noise analyses, 171, 274 Monoacylglycerol lipase, s e e Acylglycerol lipase Monocarboxylate transport in red blood cells, kinetics and chemical modification, 173. 300 Monocarboxylate transporter substrate specificity, 173, 319 Monoclonal antibodies adenosine deaminase-binding protein characterization, 178, 172 in ld-anti-Id immunoassays, 178, 174 modification, 178, 174 production, 178, 172 purification, 178, 174 advantages and disadvantages, 182, 671 in analysis of Ca2÷-bindingproteins on platelet surface, 169, 368 vasopressin, 168, 574 anti-GAT, conformational idiotypic specificities on, detection, 178, 147 -antigen interactions, protein footprinting analysis, 178, 746 anti-idiotypic in analysis of cholera toxin B subunit ligand-binding domain, 178. 165 to antialprenolol antibody production and characterization, 178, 272 response in mice, 178, 272 for anti-idiotype therapy of lymphoid tumors, production, 178, 351 to antimorphine antibodies, production, 178, 232 to antischistosomal antibody associated idiotypic cascade and isotypic selection, 178, 396 for human immunoprophylaxis of schistosomiasis, 178, 402 production, 178, 392

Monoclonal antibodies description, 182, 678 generation with complementary peptides, 178, 70 hemitoxin- or holotoxin-modified, in treatment of B cell neoplasms, 178, 356 insulin receptor, anti-DM Id biological properties, 178, 307 production, 178, 303 opioid receptor, production and characterization, 178, 243 reovirus type 3 receptor development and characterization, 178. 329 structural mimicry of viral hemagglutinin, 178, 333 vaccines against poliovirus, canine parvovirus, and rabies virus, 178, 375 anti-PADGEM, preparation, 169, 312 arachidonate 5-1ipoxygenase, preparation, 187, 341 biotin production and characterization, 184, III purification from ascites fluid, 184, ll7 biotin-labeled, precipitation, 184, 518 biotinylation, 184, 346, 472 12CA5 and 9El0, for surveillance of epitope-tagged proteins, 194, 510 cell-specific, in isolation of renal epithelia, 171, 581 definition of B-cell determinants in hepatitis B antigen pre-S(2) region, 178, 639 dinitrophenylation, 184, 348 DM idiotype, production, 178, 306 epitope specificities, analysis, 184, 507 haptoglobin, in hybridoma cultures, ELISA, 182, 712 histone, preparation from immunized mice, 170, 262 nonimmunized autoimmune mice, 170, 262 to hnRNP proteins in characterization of hnRNP particles, 181, 323 preparation, 181, 320 specificity, 181, 322

160 human carbohydrate-specific, production in vitro, 179, 299 neuropathy-associated ELISA, 179, 297 IgM reacting with gangliosides, 179, 294 IgM reacting with myelin-associated glycoprotein, 179, 291 thin-layer chromatography, 179, 298 in hybridoma culture supematant, biotinylation, 184, 472 idiotypes on, detection with immunoblotting, 178, 144 idiotypic generation with complementary peptides, 178, 70 molecular modeling, 178, 103 sequence determination with Ig mRNA, 178, 94 for small ligands in anti-idiotype inhibition immunoassay, 178, 61 ascites containing, electrophoresis, 178, 56 immunoassays, 178, 51 isoelectric focusing and direct blotting, 178, 56 production, immunization methods for, 178, 49 radiolabeling, 178, 60 for immunoprecipitation of small RNPs, preparation, 180, 472 induced mobility shifts, in detection of protein in complexes, 181, 279 keratin, description, 190, 97 -mutant diphtheria toxin conjugate immunotoxins ADP-ribosylation assay, 178, 414 cytotoxicity assay, 178, 415 inactivation of protein synthesis, kinetics, 178, 418 synthesis and purification, 178, 411 -peptide interactions, size exclusion HPLC analysis, 178, 123 preparation, 182, 670 screening assays, 182, 671 single-chain, production via genetic engineering, 182, 679 subclasses, characterization, 182, 677 trinitrophenylation, 184, 347

161 Monocytes counting on chemotaxis membranes, 169, 241 HL60 cell differentiation into, induction by gangliosides, 179, 538 human, superoxide radical generation, assay, 186, 586 platelet-derived growth factor-induced chemotaxis assay, 169, 236 checkerboard analysis, 169, 241 Monoglycosylceramides perbenzoylation, 172, 544 Monosaccharides electroneutral, accumulation by Rhodotorula, energetics, 174, 641 glycoconjugate high-performance anion-exchange chromatography, 179, 76 sequence, binding position determination, 193, 591 ions, preparation for tandem MS, 193, 609 Monosialogangliosides reversed-phase HPLC, 172, 552 MONTECARLO program extension for detection of pseudoknots and hairpins in RNA, 183, 311 Morphine anti-idiotypic antibodies monoclonal, production, 178, 232 polyclonal assay, 178, 228 production, 178, 227 purification and properties, 178, 234 Mosquito isolation of Malpighian tubules, 191, 170 Mouse as model for cancer chemoprevention by retinoids, 190, 395 vitamin A-deficient, preparation, 190, 229 MSC program multiple sequence comparison, 183, 443 Mucin human tracheobronchial association with lipids, 179, 17 characterization, 179, 3 isolation, 179, 6, 15 purification, 179, 11 related sugar chains, quantitative con-

Multienzyme complexes version to radioactive oligosaccharides, 179, 261 structural analysis by N-[3H]acetylation, 179, 27O Muconate cycloisomerase assay, 188, 127 properties, 188, 129 purification from Pseudomonas putida. 188, 128 Muconolactone isomerase assay, 188, 131 properties, 188, 132 purification from Pseudomonas putida. 188, 132 Mucosa fundic, cell dispersion, 171, 485 gastric canine, endocrine cells isolation and primary culture, 192, 176 peptide release studies, 192, 184 dissociation into individual cells, 171, 445 intact, intracellular potential and resistance measurements, 192, 86 ion transport, electrophysiological studies, 192, 82 isolated effects of cimetidine and omeprazole, 191, 724 preparation, 191, 723 responsiveness to secretagogues, 191, 723 isolated cells intracellular potential and resistance measurements, 192, 91 purification, 171, 450 viability assessment, 171, 456 lumen-tubular cell pathway, resistance, 171, 624 perfusion, 191, 644 rabbit, vesicle preparation, 192, 153 transepithelial electrical measurements, 192, 83 intestinal, postischemic, leukocyte infiltration assay, 186, 729 Mucus human lung, isolation of tracheobronchial mucin, 179, 6 Multienzyme complexes existence criteria, 182, 545

Multienzyme complexes isolation problems, 182, 540 in nucleotide biosynthesis, purification, 182, 542 purification, 182, 539 reconstitution, 182, 550 Multiple indicator dilution in analysis of renal kinetics, permeation, and transport in vivo, 191, 34 Mung bean microsomes, purification of N-acetylglucosaminyltransferase II, 179, 473 glucosidase I, 179, 457 glucosidase II, 179, 462 mannosidase I, 179, 465 mannosidase II, 179, 469 Muscarinic acetylcholine receptors characterization with amiloride, 191, 747 Muscle cells, see Myocytes chloride channels, blockers, 191, 800 contraction, related coupling rules, 171, 160 fibers, stoichiometric ratio of pumped Na + and K ÷, 173, 707 net Na ÷ extrusion and K ÷ accumulation, 173, 697 skeletal, see Skeletal muscle smooth, see Smooth muscle Mutagenesis cloned suppressor tRNA gene in vitro, 181, 382 elevation in superoxide dismutasedeficient bacterial mutants, 186, 646 nondirected, in analysis of RNA splicing in Escherichia coli, 181, 523 saturation, with mixed oligonucleotides and uracil-containing MI3 templates, 185, 599 Schizosaccharomyces pombe, 194, 805 shuttle, in yeast, bacterial transposons for, 194, 329 site-directed, generated proteins conformational comparison with nonmutant proteins, 177, 74, 282 incorporation of isotopically labeled amino acids, 177, 77 Ty-induced, in Saccharomyces cerevisiae, 194, 342

162 in yeast cell culture growth conditions after mutagen treatment, 194, 274 classical methods, 194, 273 EMS- and MNNG-induced, 194, 277 mutagen selection and dose, 194, 273 by transformation with synthetic oligonucleotides, 194, 362 UV-induced, 194, 278 in vitro methods, 194, 302 Mutants arachidonate-CoA ligase-defective cell lines, production, 187, 241 bacterial, superoxide dismutase-deficient in assay of superoxide radical role in mutagenicity, 186, 647 elevated mutagenesis, 186, 646 Chlorella transport, isolation, 174, 398 diphtheria toxin coupled to monoclonal antibodies, derived immunotoxins ADP-ribosylation assay, 178, 414 cytotoxicity assay, 178, 415 inactivation of protein synthesis, kinetics, 178, 418 synthesis and purification, 178, 411 generation, 178, 410 Escherichia coli

defective in tRNA processing analysis, 181, 398 isolation, 181, 395 for minimization of proteolysis clpA, 185, 126 htpR, 185, 125 Ion, 185, 120 produced by saturation mutagenesis identification and subcloning, 185, 603 sequence subcloning, 185, 608 Saccharomyces cerevisiae

cell and nuclear fusion assays, 194, 776 with bilateral block, isolation, 194, 782 complementation and dominance tests, 194, 780 genetic screens and selections, 194, 774 isolation, 194, 787, 790

163

Mutations mnn

detection, 185,441 glycosylation defects, 185,452 isolation, 185, 446 mannoprotein isolation, 185,455 mannosyltransferase reactions, 185, 468 mnn2 transformants, isolation, 185, 448 mnn7 class, isolation, 185, 451 multiple, construction, 185, 451 N-linked oligosaccharides, isolation, 185,457 NMR characterization, 185, 457 properties, 185, 443 spt3, for elimination of chromosomal Ty transpositions, 194, 360 Ty-induced, genetic characterization, 194, 352 tRNA processing characterization, 181,415 suppressor generation, 181, 380 in vitro analysis, 181, 383 in vivo analysis, 181, 388 yeast allele identification, 194, 317 enrichment methods, 194, 276, 279 generation from cloned genes by in vitro mutagenesis and plasmid shuffling, 194, 302 mitochondrial revertants and pseudorevertants, 181, 546 splicing-defective, types, 181, 542 nonmating, mating of, 194, 91 nuclear, affecting mitochondrial splicing revertants, 181, 557 screening, 181, 553 protease-deficient biochemical analysis, 194, 444 genetic analysis, 194, 435 Rec, isolation, 194, l l9 sec, radiolabeling for protein transport analysis, 194, 669 spol3

Rec mutation identification and analysis, 194, ll7

recovery systems, 194, 111 segregation gene identification and analysis, 194, 120 spore wall, isolation, 194, 121 sugar transport, analysis, 174, 617 temperature-sensitive RNA polymerase II, transcription inhibition. 194, 418 Mutation data matrix applications, 183, 333 limitations, 183, 341 Mutations A49, C5 protein with, preparation, 181. 575 in analysis of RNA interactions with RNA and proteins, 180, 512 yeast meiosis, 194, 99 fusion, isogenic yeast MATa and MATa strains containing, 194, 780 Lon proteases, secondary phenotypes. elimination, 185, 121 at MAT, HMR, and H M L in yeast, identification, 194, 103 mitochondrial DNA isolation, 194, 150 nuclear suppressors, 181, 551 sequence analysis, 194, 160 nondirected, causing splicing defects, isolation, 181, 522 nucleotide substitutions synonymous and nonsynonymous, estimation, 183, 544 total number, estimation, 183, 531 parallel, effects on sequence distance statistics, 183, 511 point, effects on large intron of fl-globin gene, 181, 45 Rec-, identification and analysis with spol3 mutant, 194, 117 regulating yeast entry into meiosis. identification, 194, 104 reverse, effects on sequence distance statistics, 183. 511 suppressor for analysis of RNA interactions, selection in vivo, 180, 515 effects on tRNA processing in yeast. 181, 377

Mutations in tRNA processing-related genes biochemical consequences, 181, 415 genetic analysis, 181, 417 identification, 181, 401 Ty-induced, in yeast chromosomal manipulations, 194, 356 cloning, 194, 357 LYS2 and LYS5, 194, 353 Myasthenia gravis autoimvnune response in, inhibition, 178, 450 clinical, idiotypic networks in, 178, 441 experimental autoimmune analysis and treatment with antiidiotypic antibodies, 178, 433 anti-AChR response, modulation, 178, 444 idiotypic networks in, 178, 441 regulation, 178, 444 Mycobacterium M. aoium, isolation of glycopeptidolipids

and haptenic oligosaccharides, 179, 221 M. gastri MBI9, purification of 3-hexulose-6-phosphate synthase, 188, 399 M. kansasii, purification of trehalosecontaining lipooligosaccharides and constituent oligosaccharides, 179, 225 M. leprae, infected armadillo tissue, purification of phenolic glycolipid I, 179, 227 M. szulgai, purification of trehalosecontaining lipooligosaccharides and constituent oligosaccharides, 179, 226 polycyclic aromatic hydrocarbon-degrading biodegradation experiments, 188, 149 growth and culture conditions, 188, 149 isolation, 188, 148 pyrene ring oxidation, enzymatic mechanisms, 188, 151 Mycoplasma contamination of renal cell lines, detection, 191, 430 Myelin associated glycoprotein, human IgM

164 monoclonal antibodies reacting with, review, 179, 291 Myeloperoxidase activation of xenobiotics, 186, 579 in assay of leukocyte infiltration of postischemic intestinal mucosa, 186, 729 -oxidase, generation of cysteine thiyl radicals, 186, 325 Myocardium preservation during heart transplantation, role of oxygen scavengers, 186, 742 Myocytes, see also Muscle cardiac, calcium-tolerant isolation from adult rat, 173, 662 viability assessment, 173, 670 cardiac, embryonic beating in culture, monitoring, 173, 655

chicken, preparation and cultivation, 173, 637 glycoside receptors number determination, 173, 643 occupation, correlation with inhibition of active Na+,K * transport, 173,649 Na+,K*-ATPase characterization, 173, 639 effects of ouabain and low K + levels, 173, 651 inhibition, correlation with glycoside receptor occupation, 173, 649 number determination, 173,643 relationship to positive inotropic action of glycosides, 173, 655 cardiac, neonatal beating in culture, monitoring, 173, 655 glycoside receptors, number determination, 173,643 Na+,K+-ATPase characterization, 173, 639 number determination, 173, 643 rat, preparation cultivation, 173, 637 Myoglobin oriented crystals, solid-state 2H NMR, 176, 382 role in iron metabolism, 186, 34

165

Nervous system

Myosin ATPase site biotinylation, 184, 412 visualization, 184, 409 Myosin St ATPase catalyzed ~80 exchange, detection by NMR, 176, 352 Myristate and DMPC, vesicles, spontaneous insertion of membrane proteins, 171, 258 N-Myristoylation protein amino-terminal residues, 182, 640

N NADH (reduced nicotinamide adenine dinucleotide) oxidation, in assay of superoxide dismutase, 186, 209 NADH:rubredoxin reductase component of P s e u d o m o n a s o l e o v o r a n s hydrocarbon monooxygenase system, isolation, 188, 6 NADPH (reduced nicotinamide adenine dinucleotide phosphate) coupled oxidation, 186, 520 oxidation, in assay of superoxide dismutase, 186, 209 NADPH-cytochrome-c reductase yeast endoplasmic reticulum, assay, 194, 658 NADPH-cytochrome P450 (c) reductase assay, 187, 376 NAD(P)H dehydrogenase (quinone) assay, 186, 292 biological significance, 186, 297 occurrence and intracellular distribution, 186, 288 properties, 186, 293 purification, 186, 290 Naphthalene dioxygenase ferredoxin component assay, 188, 49 purification, 188, 50 iron-sulfur protein component assay, 188, 47 purification, 188, 51

reductase component assay, 188, 48 purification, 188, 49

1,4-Naphthoquinones half-wave reduction potentials, substituent effects, 186, 187 HPLC with electrochemical detection. 186, 180 National Biomedical Research Foundation data bank alignment, 183. 469 Protein Sequence Database, s e e Protein Identification Resource Natural killer cells activity, assessment, 190, 272 Needleman-Wunsch algorithm extension to three-way alignment, 183, 367 and gap penalties, in protein sequence comparison, 183, 405 gap penalty implementation, 183, 369 in implementation of hierarchical method for sequence alignment, 183, 456 for pairwise comparisons, 183, 366 programming considerations, 183, 372 Neoantigens for analysis of mycobacterial glycolipids, synthesis, 179, 239 Neocarzinostatin, s e e Zinostatin Neoglycoproteins avidin-biotinylglycan applications, 184, 658 preparation and properties, 184, 656 Neoplasms B-cell, treatment with idiotype-specific irnmunotoxins, 178, 356 Neovascularization in bioassay of arachidonic acid metabolites, 187, 382 Nepoviruses satellite RNAs, self-cleaving sequences, compilation, 180, 547 Nerves peptide-containing, characterization. 168, 800 Nervous system autonomic, s e e Autonomic nervous system central, s e e Central nervous system neuropeptide gene transcription, assay, 168, 681

Nervous system neurotensin binding assay in sectioned tissue, 168, 464 specific sites, distribution, 168, 475 tissue membranes, 168, 464 Networks idiotypic, see Idiotypic networks Neuroendocrine system diffuse binding site analysis, 168, 804 components, localization, 168, 792 cytochemical analysis, 168, 791 peptide gene expression, demonstration, 168, 802 Neurohypophyseal system in situ hybridization studies, data analysis, 168, 823 Neurokinin A mRNA, differentially spliced, assay, 168, 634 Neurons chloride channels, blockers, 191, 799 connectivity markers, combination with in situ hybridization histochemistry, 168, 778 Neuropathy associated human monoclonal antibodies, properties and analysis, 179, 291 Neuropeptide s in central nervous system, effects on autonomic nervous system, 168, 432 effects on somatostatin release from hypothalamus, 168, 728 gene transcription in nervous system tissue, assay, 168, 681 mRNA, in situ hybridization histochemical detection, 168, 702 posttranslational modification, FAB/MS analysis, 168, 72 precursors, FAB/MS analysis, 168, 90 Y, radioimmunoassay, 168, 440 Neurophysins based affinity chromatography of neurohypophyseal peptides, 168, 585 N e u r o s p o r a crassa

nuclei, isolation, 194, 743 plasma membranes, everted vesicles isolation from cell wall-bearing strains, 174, 672 cell wall-less strains, 174, 669 transport function, assay, 174, 667

166 proton-potassium symport, analysis, 174, 654 proton translocation by H+-ATPase, assays, 174, 668, 674 spherocytes, handling, 174, 655 vacuoles, intact isolated, arginine transport, 174, 515 Neurotensin fragments and analogs, characterization, 168, 465 ~H-labeled binding sites in central nervous system, distribution, 168, 470 peripheral nervous system, distribution, 168, 475 rat brain, characteristics, 168, 465 characterization, 168, 465 Neurotensin receptors radioligand binding assays, 168, 462 Neurotransmitter receptors salivary gland coupling to secretory events, 192, 30 radioligand binding assay, 192, 29 Neurotransmitters effects on somatostatin release from hypothalamus, 168, 728 exocrine activation, patch-clamp studies, 171, 663 Neutralization effects on omeprazole and SCH 28080 inhibition of gastric acid secretion, 191, 731 immuno-, see Immunoneutralization Neutrophils collection and processing, 187, 587 counting on chemotaxis membranes, 169, 241 12S-20-dihydroxyeicosatetraenoic acid production, assay, 187, 594 effects on endotoxin-induced circulatory shock, 186, 661 eicosanoid interactions, 187, 585 human activated, oxidized proteins, assay, 186, 575 cellular integrity measurement, 187, 563 interaction with erythrocytes: leukotriene B4 synthesis, 187, 553

167 vascular cells: leukotriene C4 synthesis, 187, 559 isolation, 186, 570, 580; 187, 556 mediated macromolecular binding, assay, 186, 581 PAF-induced aggregation in vitro: analysis of WEB 2086 and 2170, 187, 459 preparation, 186, 576; 187, 559 stimulated, activation of xenobiotics, 186, 579 superoxide production, kinetic microplate assay, 186, 567 -mast cell interactions: leukotriene m4 release and metabolism, 187, 567 membranes, phospholipase C-catalyzed inositol trisphosphate production, assay, 187, 232 perfused hearts, eicosanoid and plateletactivating factor studies, 187, 615 platelet-derived growth factor-induced chemotaxis assays, 169, 236 checkerboard analysis, 169, 241 preparation, 187, 569 role in ischemia-reperfusion injury of intestine, 186, 729 reperfusion injury, 186, 714 suspensions diisopropyl fluorophosphate treatment, 187, 596 for studies of eicosanoid metabolism, preparation, 187, 593 Nicotiana, see Tobacco Nicotinic receptors affinity purification with anti-idiotypic antibodies, 178, 190 Night blindness as measure of vitamin A status in human populations, 190, 246 Nitella translucens

light-induced hyperpolarization demonstration, conditions for, 174, 483 dependence on ATP hydrolysis, 174, 487 electrophysiology, 174, 480 Nitrenes general properties, 172, 635

Nitroxide Nitric oxide chemiluminescence assay, 186, 610 N-6(7-Nitro-2, 1,3-benzoxadiazol-4-yl) ceramide labeling of MDCK cells, 191, 816 3-Nitrobenzyl alcohol mass spectra, 193, 879 Nitro blue tetrazolium reduction, in assay of HL60 cell differentiation, 190, 122 Nitrocellulose adsorbent for sample application in plasma desorption MS, 193, 434 blots, in affinity purification of antibodies, 194, 581 replicas, proteins on, staining, 184, 433 Nitrofurantoin reaction with thiols, 186, 325 Nitrogen assignment in heteronuclear NMR, 176, 108 -carbon correlations in 2D NMR of proteins, 176, 46 liquid, filtration of maize leaf material under, 174, 543 ~5N incorporation into protein side chains. 177, 59 labeling of proteins for ~H and ~N NMR, 177, 44 selective labeling of T4 lysozyme. 177. 60 3 '-(4-Nitrophenylcarbinol)-2' ,4' ,6' .4-tet rahydroxy-dihydrochalcone synthesis, 191, 757 4-Nitrophenyl chloroformate activation of Sepharose CL-4B, 184, 81. 187 2-Nitrophenyl octyl ether mass spectra, 193, 880 4-Nitrophloretin synthesis, 191, 761 4-Nitrophlorizin synthesis, 191,762 Nitrosoguanidine mutagenesis of Schizosaccharomyces pombe, 194, 806 Nitroxide reduction in dermal homogenates, analysis, 186, 672

NMR NMR, s e e Nuclear magnetic resonance NOE, s e e Nuclear Overhauser effect Noise analysis ion carriers in planar bilayers, 171, 274 Nonelectrolytes permeability of isolated perfused renal tubules, 191, 368 permeators for passive permeability measurements, selection, 174, 251 transport in red blood cells and ghosts, assay, 173, 160 light scattering studies, 173, 268 Nonidet P-40 prevention of loss of rRNA terminal maturase activity in solution, 181, 370 Noradrenaline, s e e Norepinephrine Norepinephrine hemorrhage-induced release in supraoptic nucleus, microdialysis assay, 168. 201 plasma assay, 168, 435 turnover assessment, 168, 439 Northern blot analysis interstitial retinol-bindingprotein mRNA from retinoblastoma cells, 190, 147 oligodeoxynucleotide/RNase H digestion products, 181, 291 oxytocin mRNA from brain, 168, 405 3~P-labeled ssDNA as probe, 168, 403 retinoic acid-regulated genes expressed by murine teratocarcinoma cells, 190, 134 retinoid-binding protein mRNA, 190, 153 vasopressin mRNA from brain, 168, 405 yeast low-molecular-weight RNA, 194, 414 Nuclear magnetic resonance alkali metals, properties, 171, 317 in analysis of amino acid flux in red cells, 173, 137 intraceUular metabolism, 177, 437 ion interactions at membranous polypeptide sites, 171, 286 protein structure in solution, 177, 125 RNA melting transitions, 180, 322 water diffusion across red cell membrane, 173, 201

168 13C in analysis of fatty acid biosynthesis, 177, 429 labeled glycerol metabolism, 177, 421 relative fluxes through Krebs cycle, 177, 423 in assay of methyl linoleate hydroperoxides, 186, 380 pyruvate kinase flux, 177, 430 high-field, retinylidene imminium salts, 189, 112 mechanistic studies with ~80, 177, 376 in monitoring of bioenergetics, 177, 448 phospholipid metabolism, 177, 449 with off-resonance rf field, protein rotational correlation times, 176, 386 one-dimensional, labeled proteins, 177, 79 synthetic retinoic acid analogs, 189, 31, 33 caged nucleotides, 172, 297 in characterization of yeast m n n mutants, 185, 457 databases, 176, 62 detection of insensitive nuclei, 176, 134 distance geometric analysis, 177, 159 fluorine, fluorinated ligands, 177, 3 IH in characterization of protein folding intermediates, 176, 446 cholesterol 5,6-epoxides, 186, 442 computer-assisted resonance assignments, 177, 150 Fourier transform, solvent suppression, 176, 64 ganglioside G~3 derivatives, 179, 247 high-field, retinylidene imminium salts, 189, 112 instrumentation for NOE studies of ligands, 177, 340 one-dimensional, labeled proteins, 177, 79 paramagnetic metalloproteins, 177, 246 peak analysis, automation, 177, 153 proteins deuteration methods, 177, 23

169 expression and ~SN labeling for, 177, 44 resonance assignments sequential, 177, 141 stereospecific, 177, 144 retinoic acid analogs, 189, 26 spin systems identification, 177, 133, 155 sequential assignment, 177, 155 two-dimensional, oligosaccharides conformational analysis, 179, 153 primary structure determination, 179, 123 2H, solid-state, proteins, 176, 376 heteronuclear, proteins, 176, 93 heuristic refinement, in determination of protein solution structure, 177, 218 homonuclear Hartmann-Hahn, s e e Homonuclear Hartmann-Hahn spectroscopy isotope exchange processes homonuclear spin-echo experiments, 176, 345 indirect detection of labels with spin echoes, 176, 347 proton exchange, 176, 343 line shape analysis computation under influence of exchange, 176, 282 for determination of exchange rates, 176, 279 experimental considerations, 176, 281 simulated and experimental, comparison, 176, 291 macromolecules, paramagnetic probes for, 177, 403 magnetization transfer, analysis of chemical exchange reactions, 176, 311 and molecular dynamics simulation, in determination of molecular structure, 177, 204 multiple-quantum, proteins, 176, ll4 t5N mechanistic studies with ~80, 177, 376 protein expression and rSN labeling for, 177, 44 non-Fourier transform spectral estimation methods, 176, 216 nuclear Overhauser effect, s e e Nuclear Overhauser effect

Nuclear magnetic resonance nucleic acids, sample preparation, 176. 78 one-dimensional, spin-labeled proteins, 177, 95 31p adenylate kinase, 177, 365 arginine kinase, 177, 362 creatine kinase, 177, 364 in determination of biomembrane surface potential, 171, 370 electrostatic potential adjacent to membranes, 171, 352 equilibrium constants of enzymebound reactants and products, 177, 358 enzyme complexes, 177, 295 methionyl-tRNA synthetase, 177, 372 in monitoring of bioenergetics, 177, 448 phospholipid metabolism, 177, 449 mononucleotide • ribonuclease A complexes, 177, 313 3-phosphoglycerate kinase, 177, 371 phosphoproteins, 177, 263 in positional isotope exchange, 177. 390 pyruvate kinase, 177, 369 peak intensity measurements, 176, 6 proteins, sample preparation, 176, 78: 177, 79 pulse-gradient spin-echo, in measurement of translational motion, 176, 418 quadrupolar ion. relaxation methods. 171, 290 quadrupolar nuclei in analysis of ligand-protein interactions, 177, 317 experimental fundamentals and methods, 177, 318 related computer programs, availability, summaries, and critiques, 176, 477: 177, 455 related pH indicators, 172, 84 relaxation and exchange rates, related computer programs, 176, 480 internuclear, leading to 2D NOE spectrum, 176, 170

Nuclear magnetic resonance in interpretation of protein dynamics, 176, 358 matrix analysis, see Relaxation matrix analysis protons in macromolecules, 176, 363 relationship to spectral density, 176, 363 selective methods applications, 176, 197 data analysis, 176, 195 frequency-selective pulsed excitation, 176, 186 spin-lattice, selectively excited nuclei, 176, 188 retinoid binding proteins expressed in Escherichia coli, 189, 516 solid-state, protein structural studies, 176, 242 spin-labeled proteins, 177, 91 two-dimensional in enzyme kinetics in vivo, 176, 330 in metabolite detection, 176, 336 proteins cross-relaxation spectroscopy, 176, 46 data processing and storage, 176, 50 general strategies, 176, 27 instrumentation, 176, 22 J-correlated spectroscopy, 176, 28 sensitivity, 176, 24 sequential assignment methods, 176, 50 structural analysis, 176, 50 time-domain data acquisition, 176, 28 2D correlations with less sensitive nuclei, 176, 38 solvent suppression, 176, 75 spin-labeled proteins, 177, 100 three- and four-dimensional methods, 176, 59 Nuclear Overhauser effect in analysis of intermolecular protein-ligand interactions, 177, 355 ligand conformations and ligandenzyme interactions, 177, 333 correlation time estimation, 177, 337 data collection, 177, 343 data reduction, 177, 345

170 experiments in paramagnetic metalloproteins, 177, 260 instrumentation, 177, 340 interference with Hartmann-Hahn effects, 176, 159 longitudinal relaxation rate determination by selective saturation recovery, 177, 344 one-dimensional, measurements, 176, 3 rotating-frame analysis of build-up curves, 176, 215 artifactual contributions, 176, 211 cross-relaxation rates in, 176, 199 macromolecules, 176, 207 molecular motion, 176, 210 resonance-offset effects, 176, 214 small and moderately sized molecules, 176, 202 suppression of HOHAHA transfer, 176, 213 sample preparation, 177, 339 sequential NMR resonance assignments, 177, 141 staphylococcus nuclease cross-peaks, residue-type assignment, 177, 284 steady-state, theory, 177, 335 time dependence, theory, 177, 336 T4 lysozyme, 1D and 2D isotope-directed measurements, 177, 69 two-dimensional analysis of chemical exchange, 176, 319 data collection, 177, 344 structure/intensity relationship, 176, 170 Nuclear run-off assay transcription in vitro, 190, 137 Nuclear run-on assay neuropeptide gene transcription in nervous system tissue, 168, 681 Nuclease P~ in determination of RNA branch trinucleotide core composition, 181, 188 Nuclease protection assay end-labeled cDNAs in, 168, 639 -hybridization, in analysis of polyadenylation-specific events, 181, 57 RNA splicing in Escherichia coli, 181, 535

171 rRNA termini maturation in Escherichia coli, 181, 357 -solution hybridization differentially spliced substance P and neurokinin A mRNAs, 168, 634 uniformly labeled cRNAs in, 168, 645 Nuclease SI in characterization of RNA, 180, 334 digestion of DNA, 170,287 RNA-DNA hybridization reaction products, 180, 344 for RNA structure probing, specificity and properties, 180, 197 in structural probing of native snRNA, 180, 214, 221 Nucleases in analysis of chromatin structure, 170, 339 replicating chromatin, 170, 303 digestion of nuclei, 170, 284, 326 replicating chromosomes, 170, 297 transcriptionally active chromatin, 170, 317 hypersensitive sites in chromatin, analysis, 170, 269 micrococcal, see Micrococcal nuclease mung bean, for RNA structure probing, specificity and properties, 180, 197 sensitive regions of chromatin, characterization, 170, 340 specificity on DNA and chromatin substrates, 170, 292 staphylococcal encoding gene, placement under control of strong inducible promoter, 177, 76 incorporation of isotopically labeled amino acids, 177, 77 NOE cross-peaks, residue-type assignment, 177, 284 in visualization of chromatin organization at replication forks, 170, 290 Nuclei atomic, see Atomic nuclei from cell cultures, preparation, 170, 320 from chicken organs, preparation, 170, 319

Nuclei CVI cell, SV40-infected, extract preparation, 170, 46 digestion by methidiumpropyl-EDTA.Fe(II), 170, 363 nuclease, 170, 284 DNA content evaluation, 170, 322 flow cytometric analysis, 171, 561 DNA-protein crosslinking, 170, 390 Drosophila

isolated, binding of nuclear factors in vitro, 170, 286 isolation, 170,281 nuclease digestion, 170, 284 preparation from embryo, 170, 321 purification of DNA, 170, 286 erythrocyte isolation from chicken blood, 170, 434 preparation from chicken, 170, 318 genes, evolution, quantitative estimates: effect of base composition, 183, 581 HeLa cell extracts active in polyadenylation, preparation, 181, 50 active in pre-mRNA splicing, preparation, 181, 7 efficient in pre-mRNA splicing, small-scale preparation, 181, 22 fractionation, 181, 69 for immunopurification of snRNPs, preparation, 181, 239 purification of nuclear cap binding proteins, 181, 212 with RNA lariat debranching activity, preparation, 181, 183 phosphoproteins, detection and purification, 181, 29 preparation, 170, 348 and homogenates, preparation, 170, 318 isolated binding of nuclear factors in vitro, 170, 286 steroid-receptor complexes, assay, 191, 532 histone preparation, 170, 534 hnRNA, pulse labeling, 180, 82 isolation, 180, 91 pre-rRNA, methylation, 180, 107

Nuclei isolation for analysis of hypersensitive sites in chromating, 170, 281 neuropeptide gene transcription assay, 168, 683 pulse labeling of hnRNA, 180, 86 isolation from cells, 168, 621 lysis, 180, 12 mammalian cell extracts analysis, 189, 244 fractionation, 181, 87 for histone-specific RNA 3' processing, preparation, 181, 76 snRNP levels, depletion by im° munoprecipitation, 181, 83 fractionation, 189, 241 isolated, binding of nuclear factors, 170, 286 isolation for analysis of hypersensitive sites in chromatin, 170, 281 preparation of hnRNP particles, 181, 294 procedure, 182, 210 isolation of 40S hnRNP particles, 181, 294 nuclease digestion, 170, 284 preparation, 189, 241 purification of DNA, 170, 286 retinoic acid-binding proteins and receptors, identification and analysis, 189, 239 oil-isolated characteristics, 181, 40 experimental manipulation, 181, 41 preparation with detergents, 180, II by hypotonic lysis, 180, l 1 purification of DNA, 170, 286 retinoic acid receptors cloning, analysis, and function, 189, 256 isolation and binding characteristics, 189, 249 RNA flow cytometric assay, 171, 561 isolation, 180, l0 Saccharomyces cerevisiae fractionation, 194, 746 isolation, 194, 735

172 Schizosaccharomyces pombe, staining, 194, 821 sea urchin embryo functionally intact, isolation, 170, 435 isolation with detergent, 170, 437 sea urchin sperm, isolation, 170, 438 spreading, in visualization of yeast synaptonemal complexes, 194, 127 transcriptionally active nonaqueous isolation from Xenopus oocytes, 181, 36 preparation, 181, 30 transcription in vitro, in pulse labeling of hnRNA, 180, 86 Xenopus kidney cell, preparation from culture, 170, 348 yeast DNA movement during mitosis, digital imaging microscopy, 194, 750 electron microscopy, 194, 130 extract preparation, 194, 546 genes involved in mitochondrial splicing, screening, 181, 553 immunofluorescent detection, 194, 600 isolation, 170, 32 light microscopy, 194, 129 preparation, 182, 171 spread meiotic, preparation, 194, 129 staining, 194, 99 Nucleic acids constituents MS analysis, overview, 193, 771 trimethylsilyl derivatives, preparation, 193, 791 defined-length, preparation, 181, 308 hybridization probes, labeling, 184, 577 hydrolysates, nucleosides in, qualitative analysis, 193, 777 labeling with photobiotin acetate, 184, 593 -protein affinity, in purification of yeagt plasmid chromatin, 170, 35 -protein crosslinking, reagents for, 172, 604 removal from plant extracts, 182, 192 prokaryote lysates, 182, 153 protein samples, 186, 466 yeast extracts, 182, 167 sample preparation for NMR, 176, 78

173 in vitro reaction products, characteriza-

tion, 193, 780 Nucleobases damaged by radicals, gas chromatographic assay, 186, 514 Nucleohistone in identification of DNA-protein crosslinks, 186, 540 Nucleolus yeast, specific positive stains for optical and electron microscopy, 194, 717 Nucleoplasm HeLa cells extract preparation and concentration, 181, 266 preparation by cellular fractionation, 181, 321,328 Nucleoplasmin egg, in chromatin assembly in vitro, 170, 612 purification from Xenopus eggs, 170, 612 purified, desalting and storage, 170, 622 in reconstitution of nucleosome cores, 170, 579 Nucleoproteins derivatization for GC/MS, 193, 845 DNA-protein crosslinks, GC/MS-SIM analysis, 193, 855 free radical-induced products, assay, 193, 855 histone subunits, extraction, 170, 450 hydrolysis, 193, 843 isolation, 170, 433 Nucleosides characterization in simple carrier model, 173, 253 GC/MS analysis, 193, 848 natural, structure determination, 193, 772 in nucleic acid hydrolysates, qualitative analysis, 193, 777 red blood cell, transporter kinetics, 173, 254 side-chain substitution, recognition, 193, 836 thermospray mass spectra, 193, 807 transport across red cell membranes, 173, 250

Nucleosomes trimethylsilyl derivatives electron ionization MS analysis, 193. 825 preparation, 193, 792 Nucleoside triphosphates substrate specificity of T7 RNA polymerase, 180, 61 Nucleosomes alignment on DNA, 170, 594 analysis with antihistone antibodies, 170, 251 -antibody complexes fixation, 170, 186 preparation, 170, 185 -antibody interactions, gel retardation study, 170, 230 antigenic determinants, exposure, 170, 234 arcs and cylinders from purified core particles, structure, 170, 110 assembly assay, 170, 625 extent, influencing factors, 170, 626 in vitro methods, 170, 575 associated DNA, 2-D electrophoretic resolution and sequence detection. 170, 129 associated proteins, 2-D electrophoretic resolution, 170, 136 binding on H5, strong site radiolabeling, 170, 376 characterization as chromatin repeat unit, 170, 101 composition, immunocbemical analysis. 170, 230 core particles assembly from natural and reconstituted octamers, 170, 493 authenticity, DNase I footprinting analysis, 170, 580 characterization, 170, 104 crystallization, 170, 420 internal structure: DNase I sensitivity, 170, 107 large-scale preparation and analysis, 170, 112 Lys-containing DNA-binding regions on histone octamer surface, radiolabeling, 170, 374 preparation, 170, I l

Nucleosomes purified, arcs and cylinders, structure, 170, 110 pyrene-labeled, excimer formation, 170, 205 cores reconstitution, 170, 575 -transcription factor complex, reconstitution, 170, 583 digestion with DNase I and II, 170, 264 discovery by sucrose gradient analysis, 170, 97 DNA-protein crosslinking on, 170, 389 electrophoretic analysis, 170, 116 eukaryotic cell, preparation, 170, 3 evolutionary studies, 170, 229 first on arms of replication forks, distance to ends of nascent DNA, 170, 308 fluorescence analysis, 170, 192 fluorescent labeling, 170, 193 frozen hydrated suspensions, electron microscopy, 170, 174 functional studies, 170, 241 HI-containing, localization of 5-methylcytosine, 170, 248 heterogeneity, demonstration, 170, 239 immunoelectron microscopy, 170, 180 immunogens for antihistone antibodies, preparation, 170, 255 immunological probes applications, 170, 229 assay, 170, 220 epitope-specific, 170, 227 preparation and characterization, 170, 215 purification, 170, 225 internal distances, measurement, 170, 208 oligomers, preparation, 170, 10 one-dimensional electrophoretic resolution, 170, 118 140-base pair, labeling, 170, 110 organization, immunochemical analysis, 170, 230 poly-, see Polynuoleosomes positioning, analysis by indirect endlabeling and molecular cloning, 170, 4O8 positions analysis by reconstitution in vitro, 170, 630

174 exonuclease III assay, 170, 639 reconstitution effect of DNA length, 170, 636 at physiological ionic strength, 170, 593 regularly spaced, assembly in crude cell extracts, 170, 580 repeats and ladders, 170, 345 simian virus 40, electron microscopy, 170, 176 spacing in absence of prenucleosomal DNA, 170, 310 on chromatin assembled with nucleoplasmin in vitro, 170, 630 in immature chromatin, 170, 304 periodicity for poly[d(A-T)] chromatin, analysis, 170, 600 structure in absence of prenucleosomal DNA, 170, 310 in immature chromatin, 170, 304 role of histone HI, 170, 104 sucrose gradient analysis, 170, 55 5'-Nucleotidase in assay for HL60 cell differentiation, 190, 129 Nucleotide pyrophosphatases cleavage of capped mRNA pyrophosphate linkage, 180, 171 Nucleotides ambiguous, codes for sequence comparisons with FASTP and FASTA programs, 183, 98 analogs chemically cleavable biotin-labeled, synthesis, 184, 584 for RNA-protein photoaffinity crosslinking studies, 180, 394 biosynthesis, multienzyme complexes for, purification, 182, 542 biotinylated applications, 184, 576 incorporation into DNA, 184, 572 synthesis and characterization, 184, 562 caged, synthesis and properties, 172, 288 capped, identification by terminal nucleotide analysis-, 180, 161 recognition in MS analysis of trimethyl-

175 silyl derivatives of nucleosides, 193, 841 transport in Golgi complex, 174, 173 trimethylsilyl derivatives, preparation, 193, 792 Nucleotide sequence alignment consensus analysis, 183, 221 fast, algorithms for, 183, 487 and phylogeny reconstruction, unified approach, 183, 626 ancient patterns, 183, 180 arachidonate 5-1ipoxygenase cDNA, 187, 494 branching order and dates, estimation, 183, 550 cell surface receptor genes, 191, 637 comparison, aggregate similarity scores, 183, 141 consensus DNA regulatory sites, 183, 211 mRNA, requirements for translation initiation in mammalian cells, 185, 504 databases in detection of protein similarities, 183, 111 EMBL data library, 183, 23 GenBank, 183, 3 searches, 183, 99 divergence, statistical estimation, 183, 531 DNA electrophoretic detection of specific sequences, 170, 129 mononucleosomal, in determination of nucleosome positions, 170, 414 at replication forks, relationship to chromatin structure, 170, 312 genomic, protein coding regions, detection, 183, 163 group I introns, 180, 534, 536 group II introns, 180, 535, 541 HLA-A3 genes, information profiles: CpG islands, 183, 242 homologous, aligned, evolutionary tree construction from, 183,601 homology domains detection, 183, 136 reporting and display, 183, 139

Nucleotide sequence in sequence similarity detection, 183~ 133 immunoglobulin mRNA, 178, 98 multiple, simultaneous comparison, 183. 447 patterns, searches, 183, 193 phylogenetic analysis in detection of RNA pseudoknots. 180, 300 dynamically weighted parsimony method, 183, 615 maximum likelihood method, 183, 584 maximum parsimony approach, 183, 601 RNA secondary structure, 180, 227 and sequence alignment, unified approach, 183, 626 pre-mRNA, contribution to yeast splicing efficiency, 181, 122 pre-rRNAs, 180, 104 primer extension products, 180, 368 prostaglandin endoperoxide synthase cDNA, 187, 477 RNA determination by dideoxy sequencing with reverse transcriptase, 180, 121 enzymatic sequencing, 180, 154 fragments labeled at 5' and 3' ends, 181, 587 phylogenetic comparison: detection of pseudoknots, 180, 300 RNase P RNA, in phylogenetic analysis of secondary structure, 180, 227 self-cleaving, in RNA from plant virus satellite RNAs and other sources. compilation, 180, 546 sequence space, statistical geometry, 183, 505 similarity, detection with homology domains, 183, 133 snRNA, compilation, 180, 521 substitutions synonymous and nonsynonymous. estimation, 183, 544 total number, estimation, 183, 531 synthetic oligonucleotides, 193, 778 yeast DNA correlation of repeated sequences with bands in restriction spectra, 194. 175

Nucleotide sequence mitochondrial, mutations, 194, 160 by primer extension, 194, 550 yeast mRNA, requirements for efficient translation, 185, 366 Nucleotidyltransferase in 3'-end labeling of RNA, 180, 196 Nycodenz as gradient material for buoyant density cell separation, 171, 505 solutions, density determination, 171, 510 Nystatin in chloride loading of erythrocytes, 173, 438

O Octapole reaction cells in tandem MS, 193, 169 Oerskovia

purification of nonspecific mannoprotein a-mannosidase, 185, 464 Oil isolated nuclei characteristics, 181, 40 manipulation, 181, 41 isolation of oocyte nuclei under, 181, 38 Okazaki fragments release from replication forks, 170, 311 1-Oleoyl-2-acetylglycerol in assay of platelet protein phosphorylation, 169, 434 platelet secretion of serotonin and/3N-acetylglucosaminidase, 169, 435 chemical synthesis, 169, 431 synergistic action with Ca2+ ionophore in platelet secretion, 169, 436 Olfactory bulb ovine, oxytocin release, assay, 168, 199 Oligodeoxynucleotides biotinylation for visualization of neuroendocrine mRNA, 168, 753 complementary, in site-specific degradation of RNA of snRNP particles, 181, 284 primers for dideoxysequencing of RNA annealing to template, 180, 128 chain-termination reaction, 180, 129 purification, 180, 127 selection, 180, 126

176 Oligo(dT)-cellulose in affinity chromatogr.aphy of RNA, 180, 34 for poly(A)-specific fractionation by chromatography, 180, 414 Oligomerization proteins, correlation with transport, 191, 841 Oligonucleotides branched base composition, 180, 188 characterization, 180, 185 high-voltage paper electrophoresis, 180, 189 isolation from RNA labeled in vivo with inorganic [32p]phosphate, 180, 178 RNA lariats synthesized in vitro, 180, 182 48-mer, in cloning of leukotriene A4 hydrolase cDNA, 187, 489 long, in protein gene cloning from purified protein, 182, 738 MATa2, clones containing, screening, 185, 361 mixed, in saturation mutagenesis, 185, 599 mutagenic, production, 185, 602 one-dimensional analysis with polyacrylamide gels, 180, 437 for primer extension of RNA, selection and preparation, 180, 352 probes for cloning prostaglandin endoperoxide synthases, preparation, 187, 470 high-resolution in situ hybridization histochemistry specificity, 168, 765 synthesis and labeling, 168,763 ribonuclease-resistant, generation from crosslinked RNAs, 180, 431 sequence identification with FASTA program, 183, 91 short, in protein gene cloning from purified protein, 182, 738 synthetic molecular weight and sequence determination, 193, 778 transformation of yeast, 194, 362 targeted RNase H, inactivation of U7 snRNPs, 181, 85

177 Oligopeptides analysis by collision-induced dissociation of multiply charged molecular ions, 193, 428 electrospray ionization MS analysis, 193, 421 sequences, identification with FASTA program, 183, 91 Oligosaccharides affinity chromatography on Allo A-lI-agarose column, 179, 335 E4-phytohemagglutinin-agarose column, 179, 46 Asn-linked, structural studies, 193, 672 chondroitin microsomal sulfation, 179,422 properties, 179, 426 conformation, 2D ~H NMR study, 179, 153 coupled with hydrophobic chromophores desorption MS analysis, 193, 661 liquid secondary ion MS analysis, 193, 668 reversed-phase chromatography, 193, 667 derivatization, 193, 665, 693 functional role in glycoproteins, 179, 517 in glycoconjugates, structural characterization, 193, 607 glycolipid-derived, lectin affinity chromatography, 179, 30 haptenic, isolation from Mycobacterium aviurn, 179, 221 high-energy CID-MS analysis, 193, 689 high mannose, CID spectra, 193, 701 high-performance anion-exchange chromatography, 193, 517 high-performance liquid affinity chromatography, 179, 55 ion-exchange chromatography with pulsed amperometric detection, 179, 65 isolated from nicotinic acetylcholine receptor, MS analysis, 193, 679 isolation from glycoproteins, 193, 589 glycosphingolipids, 193, 590 N-linked complex, CID spectra, 193,696, 705 in mannoprotein

Onion isolation, 185,457 NMR characterization, 185,457 O-linked preparation, 193, 734 from yeast glycoproteins, autoradiography, 194, 696 peracetylated acetolysis, 193, 653 desorption MS analysis, 193, 647 high-energy CID-MS analysis, 193, 705 peracetylation in desorption MS studies, 193, 652 periodate cleavage studies 193, 589 permethylated desorption MS analysis, 193, 647 GC/MS analysis, 193, 733 high-energy C1D-MS analysis. 193. 705 methanolysis, 193, 651 permethylation in carbohydrate mapping and fingerprinting, 193, 515 desorption MS studies, 193, 647 determination of linkage positions in glycoconjugates, 193, 589 high-mass GC/MS studies, 193, 734 primary structure, 2D ~H NMR study, 179, 123 radioactive, quantitative conversion of mucin-type sugar chains to, 179, 261 reducing terminal derivatives, CID spectra, 193, 700 structure classification from specific attachment sites, 193, 512 glycosyltransferases as probes, 179, 82 tandem MS analysis, 179, 157 trehalose-containing, lipooligosaccharide-derived, purification from mycobacteria, 179, 225 underivatized, C1D spectra, 193, 695 Omeprazole effects on gastric gland preparations, 191, 729 isolated gastric mucosa, 191,724 inhibition of gastric acid secretion, effect of neutralization, 191, 731 Onion guard cell protoplasts, preparation. 174. 319

Oocytes Oocytes, see also Eggs amphibian, microinjection of antibodies, 170, 241 rat, isolation, 168, 424 Xenopus laevis

cell-free transcription extracts efficiencies, 170, 355 preparation, 170, 351 erythroid band 3 mRNA-injected, anion transport, assay, 173, 453 extracts, chromatin assembly, 170, 603 germinal vesicles isolation, 181, 513 purification of tRNA splicing endonuclease, 181,515 homogenates, preparation, 170, 353 immature, purification of transcription factor TFIIIA, 170, 357 microinjection of erythroid band 3 mRNA, 173, 456 nuclear extracts preparation, 170, 354 transcription efficiencies, 170, 355 supernatant S-150, preparation, 170, 606 transcriptionally active nuclei, nonaqueous isolation, 151, 36 unfertilized, extracts preparation, 170, 354 transcription efficiencies, 170, 355 Opiate ligands properties, 178, 223 Opiate receptors anti-idiotypic antibodies binding, 178, 240 monoclonal, production, 178, 232 polyclonal assay, 178, 228 production, 178, 227 purification and properties, 178, 234 antimorphine antibodies, properties, 178, 224 properties, 178, 222 Opioid receptors anti-idiotypic antibodies, production and characterization, 178, 243 binding assays, 178, 246 Opioids binding to rat brain membranes, analysis, 178, 256

178 solubilized receptors, inhibition, 178, 259 neural action in vitro, 168, 121 Optical probes in determination of membrane potential, 172, 72 as internal pH indicators, 172, 82 Optical spectroscopy in analysis of retinoid-phospholipid membrane interactions, 189, 373 Organ cultures fetal bone, 190, 423 human skin, 190, 211 Organelles cytoplasmic electron microscopy, 182, 223 isolation, preparation of cell homogenates for, 182, 204 intactness, determination, 182, 207 marker enzymes, assay considerations and bookkeeping, 182, 211 membrane potential, optical measurement, 172, 102 partitioning in aqueous polymer twophase systems, 171, 532 protein assay, 182, 222 proton electrochemical potential gradient in, 172, 63 subcellular, isolation from mammalian cell lines, 182, 203 yeast enrichment, 194, 666 fractionation, 194, 662 marker enzyme assays, 194, 668 membrane-bound, detection, 194, 600 Organic solutes in fluid secreted by Malpighian tubules, analysis, 191, 174 low-capacity transport systems, detection in fish renal tubules, 191, 193 in reptilian proximal tubules intracellular levels, assay, 191, 208 transepithelial transport fluxes, 191, 207 mechanism, 191, 219 Organism 4025 purification of blue copper proteins, 188, 285 Organogenesis transplacental and maternal retinoid

179 pharmacokinetics during, correlation with teratogenicity, 190, 437 Organs chicken, preparation of nuclei and homogenates, 170, 319 exposed, detection of chemiluminescence, 186, 599 perfused detection of chemiluminescence, 186, 599 isolated, hypoxia and ischemia/reperfusion oxidant stress, 186, 752 platelet distribution in vivo, radiochemical assay, 169, 172 Ornithine decarboxylase assay: retinoid modulation of phorbol ester effects in skin, 190, 348 in heterologous gene selection and coamplification in mammalian cells, 185, 556 O r y c h o p h r a g m u s violaceous

suspension culture, protoplast isolation, 181, 152 Osmium tetroxide -ferrocyanide fixation, in visualization of yeast membrane structures, 194, 607 Osmolality gradient solutions, determination, 171, 510 Osmosis chemi-, see Chemiosmosis induced red cell swelling or shrinking, in analysis of sugar transport, 173,235 Osmotic minipumps implanted in stomach, for etretinate infusion, 190,444 Osmotic permeability endoosmotic, determination, 174, 264 in Malpighian tubules, measurement, 192, 630 water in isolated renal tubule segments, 191, 249 Osmotic permeability coefficient, see Hydraulic conductivity Osmotic pressure in insect Malpighian tubules, measurement, 191, 172 systemic plasma colloid, in assessment of glomerular filtration, 191, 113, 124

Oxidation Ouabain binding to isolated nephron segments, microassay, 191, 315 Na +,K+-ATPase, in measurement of pump site density, 173, 712 whole cells in suspension, 173, 683 effect on number of sodium pump molecules in cultured heart cells, 173, 651 3H-labeled, binding by renal tubules, assay, 191, 549 sensitive membrane hyperpolarization, in assay of Na+-K + pump activity, 173, 712 Ouabain receptors in cells forming tightly adherent monolayers, assay, 173, 686 Ouchterlony double diffusion antibody detection, evaluation, 182, 668 Ovary rat, perfused, NMR studies, 177, 441 Ovomucoid commercial, purification, 182, 335 Oxanol in determination of yeast membrane potential, 174, 616 Oxidants exposure of intact cells and purified proteins, 186, 489 formation, role in human disease, 186, 59 mediated DNA damage in vivo, overview, 186, 35 role in atherosclerosis, 186, 62 Oxidant stress, see Oxidative stress Oxidase -myeloperoxidase, generation of cysteine thiyl radicals, 186, 325 Oxidation arachidonic acid by cytochrome P450 enzymes, 187, 390 auto-, see Autoxidation coupled, NADPH, 186, 520 cysteine and cystine by performic acid. 182, 598 glycoconjugates by periodate for determination of linkage positions, 193, 587 hemoglobin thiols, 186, 271

Oxidation initial ring, pyrene by Mycobacterium, enzymatic mechanisms, 188, 151 long-chain alkanes in Acinetobacter, assay, 188, 10 mass spectrometric samples by performic acid, 193, 328 NAD(P)H, based chemical system, in assay of superoxide dismutase, 186, 209 photo-, see Photooxidation proteins characterization, 186, 490 mercaptan-mediated, protein preventing, analysis, 186, 478 by performic acid, 193, 373 pyrene, demonstration with fungal lignin peroxidase, 188, 163 reducing cofactors by prostaglandin H synthase, 186, 286 -reduction free radicals, properties, 186, 93 iron: analysis of lipid peroxidation, 186, 457 tocopherols, 186, 198 Oxidative phosphorylation in isolated gastric cells, assessment, 171, 460 nonequilibriumthermodynamics, 171, 403,428 Oxidative stress assay in vivo, 186, 681 drug-induced, assay in perfused organs, 186, 759 glutathione disulfide excretion into bile as indicator, 173,531 hypoxia- and ischemia/reperfusionrelated, analysis in isolated perfused organs, 186, 752 protein degradation as indicator, 186, 485 Oxidoreductases Fe-containing, component of naphthalene dioxygenase system assay, 188, 48 purification, 188, 49 Oxine "qn-labeled, preparation for labeling of platelets, 169, 174 Oxoglutarate dehydrogenase in intact mitochondria, Ca2÷-sensitive properties, 174, 104

180 Oxonol 1V optical probe for membrane potential determination, 172, 78 5-Oxoprnlyl-peptidase cerebral, types I and II, simultaneous assay, 168, 367 serum, assay, 168, 368 6-Oxo-prostaglandin F~ capillary GC-NCI-MS, 187, 20 Oxygen active species, analysis with metal-free solutions, 186, 121 in assay of potato arachidonate 5-1ipoxygenase, 187, 297 consumption in assay of 13-cis-retinoic acid inhibition of lipid peroxidation, 190, 283 by parietal cells, determination, 192, 103 effects on red blood cell deformability, 173, 51 18O in mechanistic studies with ~3Cand 15N NMR, 177, 376 P~, detection in myosin S~ ATPase, 176, 352 radical-mediated DNA damage in presence of, 186, 511 reactive species analysis by oxystat technique, 186, 443 overview, 186, 1 phycobiliprotein fluorescence decay assay, 190, 277 phycoerythrin fluorescence-based assay, 186, 161 singlet chemical verification, 186, 604 chemiluminescence assay based on luciferin analogs, 186, 585 chemiluminescent, chemistry and sources, 186, 596 generation in peroxidase-H202-halide systems, assay, 186, 590 molecular detection during chloride peroxidase-catalyzed ethyl hydroperoxide decomposition, 186, 133 enzyme-dependent chemiluminescence, 186, 135

181 reactions in solutions, rate constants, 186, 114 role in human disease, 186, 2 Oxygenation arachidonic acid by prostaglandin H synthase, 186, 285 after ischemic injury, effects on brain, 186, 80 heart, 186, 74 intestine, 186, 70 limbs, 186, 84 organ preservation for transplantation, 186, 81 pancreas, 186, 84 in rheumatoid arthritis, 186, 83 role of endothelium, 186, 78 Oxygen electrodes in assay for acetol monooxygenase, 188, 30 acetone monooxygenase, 188, 30 alcohol oxidase, 188, 421 benzene dioxygenase, 188, 53 cyclohexanone 1,2-monooxygenase, 188, 71 cyclopentanone 1,2-monooxygenase, 188, 77 methanol dehydrogenase, 188, 203, 211 methanol dehydrogenase protein M, 188, 217 Oxygen radicals drug-dependent generation, assay in vitro, 186, 621 and hydroxyl radicals, reaction with polyaminocarboxylates, 186, 117 inorganic, reactions in solution, rate constants, 186, 115 intracellular generation in E s c h e r i c h i a coli, 186, 641 kinetics and mechanisms, 186, 93 production by photosensitization, 186, 635 pulse radiolysis studies, 186, 89 reactions in solution, rate constants, 186, 113 role in cytochrome P450 peroxygenase activity, 186, 277 disseminated intravascular coagulation, 186, 656 endotoxin shock, 186, 660

Pancreas reductive cleavage of hydroperoxides by cytochrome P450, 186, 282 reperfusion injury, 186, 711 scavengers, role in myocardial preservation during heart transplantation, 186, 742 Oxyhemoglobin absorption spectra, 186, 268 Oxynitrogen radicals organic, reactions in solution, rate constants, 186, 115 Oxyntic cells, s e e Parietal cells Oxystat technique in analysis of reactive oxygen species. 186, 443 Oxytocin mRNA, cerebral dot-blot analysis, 168, 407 nonradioactive visualization in hypothalamus, 168, 753 Northern blot analysis, 168, 405,407 preparation, 168, 400 in situ hybridization, 168, 409 solution hybridization assay, 168, 404 release from sheep olfactory bulb and substantia nigra, microdialysis assay, 168, 199

P PADGEM protein antibodies assay, 169, 314 preparation, 169, 312 assay, 169, 318 detection, 169, 322 properties, 169, 318 purification, 169, 315 PAF, s e e Platelet-activating factor Paf-acether, s e e Platelet activating factor l-Palmitoyl-2-[ 10-[4-(trifluoromethyl)diaziriniyl]phenyl]-8-[9-aH]oxadecanoyl]-sn glycero-3-phosphorylcholine synthesis, 172, 649 Pancreas acini dispersed applications, 192, 275 culture preparation, 192, 277

Pancreas perifusion, 192, 278 preparation, 192, 274 isolation, 192, 288 as second messenger models in exocrine secretion, 171, 590 secretagogue stimulation, analysis, 192, 249 arachidonate compartmentalization, 191, 685 collagenase digestion, 192, 190 ducts bicarbonate transport, analysis, 192, 269 isolated, secretion studies, 192, 266 perfusion, 192, 260 exocrine tissue, separation of islets, 192, 195 glands, isolated, secretion studies, 192, 261 ischemia-reoxygenation injury, 186, 84 islet cells, see Islet cells islets anatomical distribution and cellular organization, 192, 188 isolation from adult, 192, 189 primary culture, 192, 204 separation from exocrine tissue, 192, 195 microdissection, 192, 189 minilobules, preparation and incubation, 191, 686 rat, total RNA, purification, 180, 4 reperfusion injury, role of oxygenderived free radicals, 186, 715 secretion analysis in vivo, 192, 256 stimulation by secretagogues, 192, 247 structure and function, variations, 192, 256 whole, radiolabeling, 191, 685 Pancreatic ribonuclease digestion of internally labeled RNA, 180, 134 in mapping of RNA branches, 181, 187 -mononucleotide complex, 31p NMR studies, 177, 313 Papain cleavage of anion transporter, 173, 427 PAPA program for relating progressively aligned amino acid sequences

182 application, 183, 665 availability, 183, 669 Paper chromatography in assay of endo-/3-galactosidase releasing Gal(al---~3)Gal, 179,496 glucosidase I, 179, 456 mannosidase I, 179, 464 Papillomavirus bovine, vectors for heterologous gene expression in mammalian cells, 185, 492 Papovaviruses vectors for heterologous gene expression in mammalian cells, 185, 488 Paracellular shunt conductance in epithelia, determination, 191, 4 electrical resistance, determination, 191, 16 Paracoccus denitrificans

purification of methanol dehydrogenase M protein, 188,218 methylamine dehydrogenase, 188, 242 Paraformaldehyde platelet fixation with, 169, 151 stock solutions for immunogold labeling, 194, 625 Paramagnetic probes for NMR studies of macromolecules, 177, 403 Paraquat induced H202 formation, assay in vitro, 186, 625 Parasites, see also specific parasites anti-idiotype antibodies to, 178, 13 Parathyroid hormone biotinylation, 184, 277 chromatography, 184, 277 Parathyroid hormone receptors flow cytometric detection, 184, 279 Paratopes related idiotopes, detection by inhibition ELISA, 178, 383 Parietal cells acid production, effects of prostaglandins, 187, 512 activity, determination, 192, 103 arachidonate metabolism, 187, 505 Ca2+ metabolism, analysis with Fura-2, 192, 67

183 Ca 2÷ pumping and internal store refilling, 192, 69 Ca 2+ uptake during reloading, 192, 70 dye leakage, 192, 53 gastric cyclic nucleotide-related measurements, 191, 649 functional correlation with cAMP metabolism, 191, 659 functional measurements, 191, 647 intact, cAMP-dependent protein phosphorylation, 191,654 isolation, 191, 640 purification, 191, 646 H+ flux rates, assay, 192, 56 H+,K+-ATPase distribution, 192, 158 intracellular Ca 2., assay, 192, 60 intracellular ion activities, assay, 192, 38 intracellular potential and resistance measurements in v i t r o , 192, 91 intracellnlar signaling, role of Ca -~÷, 192, 72 isolated enrichment, 191, 733 preparation, 192, 100 isolation of H +,K+-ATPase-containing membranes, 192, 151 loading of acids, mechanisms, 192, 55 bases, mechanisms, 192, 54 2' ,7'-bis(carboxyethylcarboxy) fluorescein, 192, 50 fluorescent dyes general methods, 192, 39 toxic effects, 192, 45 membranes, ion transport assay, 192, 38 Na + fluxes and pump activity, assay, 192, 78 permeabilization, 192, 108 pH, measurement, 192, 49 preparation, 187, 505 prostaglandin E2 production, effects of gastric secretagogues, 187, 513 Parotid gland rat agonist activity of antisubstance P anti-idiotypic antibodies, 178, 296 substance P receptors, binding of antiidiotypic antibodies, 178, 292

PDGF Parsimony methods dynamically weighted, phylogeny determination, 183, 615 evolutionary, analysis of molecular phylogenies, 183, 648 maximum analysis of molecular phylogenies. 183,646 evolutionary tree construction from aligned homologous sequences, 183, 601 after progressive alignment, analysis of amino acid sequences, 183, 659 Particle analyzers electronic, s e e Electronic particle analyzers PAS, s e e p-Aminosalicylic acid Passive coupling reduction by random fractional deuteration, 177, 30 Patch-clamp studies algal cell electrophysiology, 174, 431 anterior pituitary cells identified by reverse hemolytic plaque assay. 168, 144 demonstration of messenger-mediated channel opening by single-channel current recording, 171. 667 whole-cell current recording, 171, 671 epithelial activation by hormones and neurotransmitters, 171, 663 human red cell single-channel currents. 173, 112 ion transport in gall bladder epithelium, 192, 672 gastric mucosa, 192, 92 isolated guard cell protoplasts and vacuoles, 174, 312 isolated renal tubules, 191, 299 Malpighian tubule ion channels, 191, 181 water transport in gall bladder epithelium, 192, 672 whole-cell, membrane K ÷ conductance in intestinal secretory cells, 192, 309 Patency long-term, in blood-contacting prostheses, predictive value of acute e x v i v o tests, 169, 100 PCR, s e e Polymerase chain reaction PDGF. s e e Platelet-derived growth factor

Pea Pea guard cell protoplasts, preparation, 174, 318 Pectate lyase digestion of polygalacturonic acid, 179, 567 Penicillinase -avidin conjugates, preparation, 184, 179 Pentafluorobenzyl esters formation by lipid peroxide transesterification, 186, 391 Pentagastrin effect on prostaglandin E2 production by parietal cells, 187, 513 Pentetic acid in spin trapping of free radicals, 186, 131 Pentose cycle modeling: metabolism of ~3C-labeled glycerol, 177, 421 Pepsin cleavage of anion transporter, 173, 427 Pepsin A digestion of proteins for MS analysis, 193, 370 Pepsinogen assay, 192, 131 granules isolation, 192, 166 morphology, 192, 167 isolation and identification, 192, 133 secretion from isolated gastric glands, 192, 124 Peptide N4-(N-acetyl-fl-glucosaminyl) asparagine amidase applications, 179, 515 assay, 179, 508 properties on glycoprotein substrates, 179, 511 purification from Flavobacterium meningosepticum, 179, 506 Peptide esters enzymatic hydrolysis, 193, 347 Peptide mapping by FAB/MS, 168, 85 3H-reductively methylated histones after tryptic digestion, 170, 379 one-dimensional, by SDS-PAGE, 182, 622 overview, 182, 613 prostaglandin endoperoxide synthase proteolytic fragments, 187, 484

184 proteolytic, calmodulin-binding peptides, 184, 465 tryptic, in identification of in vitro glycated sites of Cu-Zn superoxide dismutase, 179, 575 Peptides amino acid residues, mass values, 193, 888 amino groups, acetylation, 193, 328 -antibody interactions, size-exclusion HPLC analysis, 178, 122 antigenic potency, enhancement, 178, 630 -antigen system, high-density multiple, for antipeptide antibody preparation, 168, 7 arachidonate 5-1ipoxygenase-derived, sequence analysis, 187, 492 atrial natriuretic, see Atrial natriuretic peptide B-cell determinant containing, design, 178, 599 biotinylated, structural analysis, 184, 281 calmodulin-binding, proteolytic mapping, 184, 465 carrier-mediated transport from brain to blood, assay, 168,652 complementary antibody-antigen interactions, 168, 27 antiparallel and parallel, preparation, 168, 16 in antireceptor antibody production, 168, 25 • in solid-phase binding assays, 168, 23 synthesis based on consensus sequences, 168, 19 continuous-flow FAB/MS, 193, 222 cRNA-encoded, in generation of antiidiotypic antibodies, 178, 63 disulfide bonds, reduction, 193, 328 dose-response testing by hippocampal brain slice recording, 168, 129 electrospray-tandem MS analysis, 193, 178 epitopes B-cell, addition of Th-cell epitopes, 178,661 with enhanced immunogenicity, vaccines based on, 178, 659 presentation on hepatitis B core protein, 178, 669

185 size-exclusion HPLC analysis, 178, 107 FAB-tandem MS analysis, 193, 174 fingerprinting, in identification of thrombospondin, 169,264 FMDV, with added foreign Th-cell epitopes, immunogenicity, 178, 666 fragment ions, nomenclature, 193, 886 fragments, N-acetylation, 193, 478 gastrin-releasing, s e e Gastrin-releasing peptide gene expression in diffuse neuroendocrine system, demonstration, 168, 802 heparin-binding, in thrombospondin, analysis, 169, 265 high-energy CID spectra, computeraided interpretation, 193, 472 histone, as immunogens for antihistone antibodies, preparation, 170, 253 from HIV gpl60 envelope protein, overlapping studies of immunogens, 178, 627 homonuclear Hartmann-Hahn spectroscopy, 176, 159 hormones, biotinylation, 184, 275 HPLC mapping applications, 182,625 for N-terminal sequence analysis, 182, 607 hydroxylated, FAB/MS, 168, 84 isolation-associated artifacts, FAB/MS analysis, 168, 88 from malaria circumsporozoite protein, overlapping studies of immunogens, 178, 627 mass spectrometry by matrix-assisted uv laser desorption/ ionization, 193, 280 molecular weight determination, utility, 193, 354 overview, 193, 351 multiple antigenic, in production of sitespecific antibodies, 178, 739 nerves containing, characterization, 168, 800 neuro-, s e e Neuropeptides neuroendocrine gene expression, regulation, 168, 618 hnRNA, assay, 168, 626

Peptides isolation from milk and identification, 168, 494 microdialysis assay, 168, 166 mRNA, assay, 168, 626 transcription assay, 168, 622 neurohypophyseal affinity chromatography, 168, 585 conjugation to carrier protein, 168. 576 radiobinding assays, tracer effects. 168, 581 radiolabeling and tracer purification. 168, 576 nucleotide, gel electrophoresis, 170, 407 O-glycosylated, FAB/MS, 168, 82 opioid, electrophysiological analysis. 168, 103 phosphate assay, 193, 487 phosphate detection, 193, 482 phosphorylated/dephosphorylated ratio, estimation, 193, 489 phosphorylated, liquid secondary ion MS analysis, 193, 480 phosphoserine identification, 193, 492 physalaemin-related immunoreactive purification, 168, 450 structural analysis, 168, 454 poly-, s e e Polypeptides precursors, structure and processing. 168, 518 preprogastrin-releasing, translation, 168. 677 processing in vivo, evaluation, 168, 519 produced by peptidylglycine monooxygenase reaction, radioimmunoassay. 168, 355 protonated, fragmentation and keV collisions, 193, 456 -quinone adducts, o-semiquinone radicals from, characterization, 186, 339 recombinant human erythropoietinderived, identification, 193, 532 release from gastric mucosa endocrine cells, analysis, 192, 184 reversed-phase HPLC, 193, 389, 407 sequence analysis by liquid secondary ion MS. 193, 490 tandem MS, 193, 493 tandem MS and high-energy CID. 193, 455 signal, processing site determination, 185, 220

Peptides STP-2, primary structure, 193, 407 sulfate assay, 193, 487 sulfate detection, 193, 482 sulfated, liquid secondary ion MS analysis, 193, 480 synthetic antibodies, in mapping sodium channel functional sites, 178, 714 for development of antisera to procholecystokinin, 168, 4 hepatitis B antigen composite immunogen with nucleocapsid T cell and envelope B cell epitopes, 178, 654 immune response of nonhuman primates, 178, 657 priming of T-helper activity and antibody production to HBsAg, 178, 652 in immunoassay distinguishing HIV-I and HIV-2 infections, 178, 693 immunochemical antigen recognition, 178, 731 inhibin fragments, antisera, in detection and purification of inhibin, 168, 588 rhinovirus receptor, in production of site-specific antibodies, 178,677 serologic applications, 178, 694 sodium current modification by antibodies, assessment, 178, 736 thin-layer electrophoresis-thin-layer chromatography, 182, 623 tryptic doubly charged, tandem MS analysis, 193, 426 from glycogen synthase, liquid secondary ion MS analysis, 193, 483 from interstitial retinol-bindingprotein, preparation and characterization, 189, 212 from reductively methylated histones fractionation and purification, 170, 381 radiolabeled, identification and specific radioactivities, 1711, 383 tyrosine-containing, sulfated, liquid secondary ion MS analysis, 193, 486 vasoactive intestinal, s e e Vasoactive intestinal peptide

186 Peptidylglycine monooxygenase incubations, 168, 355 radioimmunoassay, 168, 351 Peracetylation oligosaccharides, 193, 589,652 Perbenzoylation glycosphingolipids with benzoic anhydride, 172, 547 benzoyi chloride, 172, 544 Perchloric acid extracts of erythrocytes, preparation, 173, 61 Percoll gradient material for buoyant density cell separation, 171, 506 nonparenchymal liver cells in, densitygradient centrifugation, 190, 66 Perfluorokerosene reference ion masses, 193, 871 Perfluorotributylamine reference ion masses, 193, 871 Performic acid oxidation of cysteine and cystine, 182, 598 mass spectrometric samples, 193, 328 proteins prior to digestion, 193, 373 Perfusion, s e e a l s o Reperfusion in analysis of platelet-surface interactions, 169, 55 chambers for, in analysis of platelet interactions with subendothelial extracellular matrix, 169, 83 vessel walls, 169, 43 gastric gland basolateral surface, 192, 42 gastric mucosa, 191, 644 giant algal cells cytoplasm, 174, 437 vacuoles, 174, 434 hippocampal slices for dose-response testing of peptides, 168, 132 internal cell, during patch-clamp studies, applications, 171, 672 intracellular, C h a r a cells, 174, 446 isolated rat heart, 186, 724 in isolation of Ca2÷-tolerant atrial and ventricular myocytes, 173, 667 renal cortical cells, 191, 381 renal medullary cells, 191, 387

187 kidney blood-free methods, 191, 131 with collagenase, 191, 227 defined nephron segments, 191, 254 from frog, 192, 713 isolated tubules, rate, 191, 241 proximal convoluted tubules, 191, 344 tubules from birds, 191, 224 elasmobranchs, 191, 184 liver enzymatic methods, 190, 62 from rat, 173, 564 Malpighian tubule lumen, 192, 627 micro-, s e e Microperfusion organ, system selection, 186, 759 pancreatic duct, 192, 260 rat lung, 187, 600 rectal gland from shark, 192, 757 tubules, 191, 183 salivary gland, 192, 10 sweat duct in vitro, 192, 591 xylem, in transport studies, apparatus, 174, 282 Perhydroxyl radicals reactions with metalpolyaminocarboxylate complexes, 186, 117 in solution, reaction rate constants, 186, 114 Perifusion human hypothalamic and pituitary tissue in vitro, 168, 206 pancreatic acini, 192, 278 pituitary cell aggregates, secretory responses during, 168, 57 system for membrane potential measurements, 192, 236 Periodate oxidation of glycoconjugates for linkage position determination, 193, 587 Permeability capillary, as index of microvascular injury, 186, 736 epithelial membranes to ions, selectivity, 171, 727 inner mitochondrial membrane to protons by mitochondrial swelling, 174, 86

Permeability constant isolated perfused renal tubules to ions, 191, 369 nonelectrolytes, 191, 368 Malpighian tubule wall analysis, 192, 623 in vitro studies, validation, 192, 625 membrane isolated gastric cells, 171,459 unstirred layers determination, validity, 173, 166 osmotic, s e e Osmotic permeability passive measurement in analysis of membrane structure. 174, 265 experimental aspects, 174, 251 principles, 174, 248 one-step inside-out vesicles, 173, 375 to solutes, measurement by endoosmotic methods, 174, 252, 262 partial concentration method, 174, 256 to water, measurement, 174, 255,263 phospholipid membranes, effects of retinoids ESR studies, 189, 391 optical spectroscopic studies, 189, 375 plasma membrane to normally impermeant compounds. alterations, 171, 858 R h o d o t o r u l a , to H + and K ÷, 174, 646 proximal tubule membrane, estimates. 191, 210 renal tubules and vesicles to water, optical measurement, 191, 561 reptilian distal tubule to water, 191, 221 transport system, application to transport system isolation, 172, 36 Permeability coefficient for isolated perfused renal tubules, calculation, 191, 368 osmotic, s e e Hydraulic conductivity solute determination from single-time course, 173, 220 water, calculation with desk-top hybrid computers, 172, 274 Permeability constant calculation, 174, 257

Permeabilization Permeabilization cells by digitonin, in determination of subcellular isotope distribution, 173, 553 to macromolecular probes by cell cracking, 168, 225 chemical methods, 192, 283 electrical methods, 192, 282 giant algal cell plasmalemma, 174, 440 mammalian cells to macromolecules, 171, 857 pancreatic acinar cells by digitonin or saponin, 192, 283,292 c~-toxin and streptolysin O, 192, 285, 296 parietal cells, 192, 108 plasma membrane by high-voltage pulses, 171, 822 pore-forming chemicals, 171, 827 red cells to Ca2+ by A23187, 173, 107, 110 transient, endocrine cells: analysis of inositol lipid metabolism, 168, 218 PERMEAR2V program modeling of solute movement and volume changes, 172, 282 PERMEA/R2V AND PLOT program modeling of solute movement and volume changes, 172, 281 Permeases high levels, yeast with, preparation, 174, 628 Permeation ionic mechanisms in epithelia, 171,678 models, 171, 696 nonelectrolyte, characterization, 171, 703 nonspecific, characteristics, 174, 567 renal, multiple indicator dilution studies in v i v o , 191, 34 Permethylation gangliosides, for MS/MS analysis, 193, 759 glycolipids, 193, 753 glycosphingolipids, 193,589 oligosaccharides for carbohydrate mapping and fingerprinting, 193, 515

188 desorption MS studies, 193, 647 determination of linkage positions in glycoconjugates, 193, 589 high-mass GC/MS studies, 193, 734 Permselectivity glomerular, multiple indicator dilution studies, 191, 49 peritubular capillary, multiple indicator dilution studies, 191, 55 Peroxidase biotinylation, 184, 474 complexes, and luminol, antigen detection, 184, 442 generation of thiyl radicals of cysteine and glutathione, 186, 319 -H202-halide systems, singlet oxygen generation, assay, 186, 590 in immunocytochemical detection of human T- and B-cell antigens, 184, 363 in oxidation of tocopherols, 186, 201 Peroxidation hydro-, s e e Hydroperoxidation lipid, s e e Lipid peroxidation methanol, in assay of drug-dependent H202 generation, 186, 622 Peroxisomes marker enzyme catalase, assay, 182, 219 yeast, immunofluorescent detection, 194, 601 Peroxyl radicals kinetics and mechanisms, 186, 96 lipid-soluble, generation by azo compounds, 186, 100 scavengers, phycoerythrin fluorescencebased assay, 186, 162 spin trapping, 186, 131 water-soluble, generation by azo compounds, 186, 100 Perthiyl radicals chemical properties, 186, 178 pH control in HCO3-/CO2solutions, 192, 42 cytoplasmic, effect on ion transport in C h a r a cells, 174, 473 effects on detergent properties, 182, 256 phenylglyoxal modification of anion transport system, 173, 473

189 superoxide dismutase inhibition, 186, 224 vesiculation of one-step inside-out vesicles, 173, 375 fluorescein as probe, 174, 132 gradients dissipation, measurement with acridine orange, 192, 403 driven C1- uptake by intestinal membrane vesicles, 192, 407 driven Na + uptake by intestinal membrane vesicles, 192, 399 formation by H+,K+-ATPase-contain ing vesicles, assay, 192, 161 in isoelectric focusing determination, 182, 473 instability, 182, 466 transmembrane, in analysis of monocarboxylate transport, 173, 316 in human red cells, modification, 173, 178 as indicator of alkali metal/proton exchange. 173, 344 indicators in determination of biomembrane surface potential examples, 171, 374 theory, 171, 377 fluorescein isothiocyanate-dextran, 174, 137 internal, 172, 82 for NMR, 172, 84 intracellular, erythrocytes varying in, preparation, 173, 57 measurement in cell cultures, 174, 140 across cell membranes, organelles, and vesicles, 174, 131 in cells of isolated nephron segments, 191, 261 in cell suspensions, 172, 87 in cytoplasm: detection of Na+/H + exchange in lymphocytes, 173, 777 in cytosol, 172, 85 in endosomes, 173, 755 external to yeast, 174, 596 gradients in cells. 172, 91; 191, 470 EPR techniques, 172, 331

Pharmacokinetics across Rhodotorula plasma membrane, 174, 632 in vesicles, 191, 470 intercellular, microelectrode techniques, 174, 331 intracellular Amphiuma erythrocytes, 173. 333 compartments in living cells, 172, 85 microelectrode techniques, 174, 331 parietal cells, 192, 49 Rhodotorula, 174, 632 yeast, 174, 596 in isolated organelles, 174, 146 in lysosomes, 173, 755 in membrane vesicles formed in vitro. 174, 153 in single identified cells, 172, 89 transmembrane differences, 172, 79 in vacuoles of living cells, 173, 745 3tp NMR titration curves for phosphoamino acids, 177, 274 yeast, effect on cation uptake, 174, 608 Phadebas amylase test in analysis of secretagogue stimulation of dispersed pancreatic acini, 192. 249 Phagocytes response to retinoids, 190, 269 superoxide production, kinetic microplate assay, 186, 567 Phagocytosis immune, in assay for HL60 cell differentiation. 190, 128 Phagosomes pH and proton pumping, assay in cell culture, 174, 145 Phalloidin fluorochrome-conjugated, staining of actin, 194, 729 Phanerochaete chrysosporium purification of lignin peroxidase, 188, 166 Pharmacodynamics embryonic retinoic acid derived from maternally administered retinol. 190. 320 Pharmacokinetic s retinoids in evaluation of developmental toxicity, 190, 406

Pharmacokinetics during organogenesis, correlation with teratogenicity, 190, 437 overview, 190, 291 Pharmacology biochemical, inflammatory hepatic injury in mice, 186, 675 ergothioneine, 186, 316 Phase diagrams nucleosome core particles, in formulation of crystallization strategies, 170, 423 Phase partitioning cells and organelles in aqueous polymer two-phase systems, 171, 532 paramagnetic labels, quantitation, 172, 334 in preparation of plasma membranes, 172, 12 proteins from exocytic transport vesicles, 191, 821 in subcellular fractionation of enterocytes, 192, 348 in subcellular membrane isolation, 172, 23 o-Phenanthroline chelatable copper, assay, 186, 40 inhibition of transcription in yeast, 194, 418 Phenol acidic, extraction of RNA, 180, 78 and chloroform, extraction of DNAprotein complexes, 170, 394 extraction of high-molecular-weight yeast RNA, 194, 399 oxygen atoms, acid-catalyzed exchange with H21sO, 193, 346 Phenoxyl radicals kinetics and mechanisms, 186, 97 Phenylalanine spin system identification, 177, 138 Phenylglyoxal exofacial modification of resealed ghosts, 173, 469 modification of anion-transport system, 173, 466 reaction with arginyl residues, 173,467 reversible inhibition of CI- exchange, 173, 476 Pheromones production by yeast, assay, 194, 84 yeast response, assay, 194, 86

190 Phloem transport loading, 174, 288 over long distances, 174, 298 unloading, 174, 308 uptake of [14C]sucrose, 174, 289 Phloretin photoaffinity-labeling analogs, synthesis and effects on cell membranes, 191, 755 Phloridzin membrane binding, renaturation after Na+-D-glucose cotransporter solubilization, 191, 590 photoaffinity-labeling analogs, synthesis and effects on cell membranes, 191, 755 Phlorizin, s e e Phloridzin Phorbol esters pretreatment of pituitary cells, 168, 288 Phosphatase in 5' labeling of DNA primers, 180, 356 Phosphate buffer stock solutions, 182, 34 -citrate buffer stock solutions, 182, 32 -H + cotransport system, repression and derepression studies, 174, 608 indicator for analysis of transport in isolated renal cells, 191, 404 initial transport rates, direct determination, 173, 737 inorganic in goblet cell apical membrane, assay, 192, 614 phosphorylation of sodium pump active site, assay, 173, 688 regulation of cloned gene expression in yeast, 194, 385 low-resolution gels, preparation, 170, 555 -phosphate exchange, N-ethylmaleimide-sensitive, assay, 173, 736 32P-labeled intraliposomal, assay, 173, 738 labeling of newly synthesized RNA, 150, 72 in proteins or peptides assay, 193, 487 detection, 193, 482 transport in renal epithelial cell lines, 191, 494

191 Phosphate transporter covalent modification with N-ethylmaleimide, 173, 738 purification from rat liver mitochondria, 173,732 purified effects of covalent labeling agents, 173,742 properties, 173,739 reconstitution, 173, 732 Phosphatidic acid disposition on stimulation of phosphatidylinositol breakdown, 191,688 platelet, endogenous assay by fatty acid quantitation, 169, 426 extraction, 169, 421 phosphorus assay, 169,426 TLC, 169,422 Phosphatidylcholine t4C-labeled equilibration between membrane leaflets, 173,227 introduction into intact erythrocytes, 173, 226 donor vesicles, preparation, 173, 225 enzymatic hydroperoxidation, 186, 453 positionally defined compounds, preparation, 189, 452 -retinol carrier vesicles, preparation, 190, 15 specific transfer protein, preparation, 173, 226 transbilayer mobility in red blood cell, 173, 223 Phosphatidylcholine hydroperoxide HPLC, 186, 308 in human plasma, HPLC assay, 186, 159 Phosphatidylinositol breakdown, subsequent disposition of moieties in, analysis, 191, 685 derived arachidonic acid, metabolism and function, 191, 676 hydrolysis in isolated cell membranes, assay, 168, 338 by phospholipase C, 187, 206 platelet, endogenous assay by fatty acid quantitation, 169, 426 extraction, 169, 421

P h o s p h o l i p a s e A2 phosphorus assay, 169, 426 TLC, 169, 422 turnover in bone marrow-derived mast cells, 187, 517 stimulated by ligand binding to bombesin/gastrin-releasing peptide receptors, 168, 490 Phosphatidylinositol 4,5-bisphosphate hydrolysis in isolated cell membranes, assay, 168, 338 platelet, endogenous assay by fatty acid quantitation, 169, 426 extraction, 169, 421 phosphorus assay, 169, 426 TLC, 169, 422 6-Phosphogluconate dehydrogenase

Arthrobacter globiformis assay, 188, 343 properties, 188, 344 purification, 188, 343

Methylobacillus flagellatum assay, 188, 337 isoenzymes, partial purification, 188, 338 properties, 188, 338 Phosphoglycerate kinase promoter, high-efficiency yeast expression vector based on, 185, 329 3-Phosphoglycerate kinase bound reactants and products, equilibrium constant determination, 177, 371 Phosphoglycerides arachidonate-containing, separation and characterization, 187, 157 Phosphoinositide cascade relationship to Ca2+ dependency of phospholipase A2, 191,682 Phosphoinositides in hepatocytes, assay, 173, 534 hydrolysis in isolated cell membranes, assay, 168, 338 phospholipase C-catalyzed, in suspension, assay, 187, 230 Phospholipase A2 Caz+ dependency, relationship to phosphoinositide cascade, 191, 682

Phospholipase A2 macrophage, sn-2-arachidonic acidspecific assay, 187, 216 partial purification, 187, 223 properties, 187, 224 role in arachidonate liberation from phospholipids, 191, 678 Phospholipase C activity, role of arachidonate and metabolites, 191, 683 catalyzed inositol trisphosphate production by neutrophil membranes, assay, 187, 232 hydrolysis of phosphoinositides in suspension, assay, 187, 230 phospholipids, 187, 206 phosphoinositide-specific, assay, 187, 226 role in arachidonate liberation from phospholipids, 191, 677 Phospholipases red cell incubation with, 173, 227 Phospholipid hydroperoxide glutathione peroxidase characteristics, 186, 448 purification from porcine heart, 186, 455 spectrophotometric assay, 186, 453 in tissues, assay, 186, 455 Phospholipid hydroperoxides production and extraction, 186, 453 Phospholipids arachidonate liberation, mechanisms, 191, 677 artificial membranes, surface potential determination, 171, 342 chromatography, 191, 687 classes, separation, 187, 198 coating of erythrocyte cytoskeletons, 173, 394 extraction from pancreatic tissue, 191,686 tissues, 187, 195 HPLC, 172, 560 LC/MS analysis, 172, 570 membrane coated cytoskeleton characterization, 173, 395 preparation by erythrocyte reconstitution, 173, 392

192 derived molecules, synthesis in bone marrow-derived mast cells, 187, 514 interactions with retinoids ESR studies, 189, 383 optical spectroscopic studies, 189, 373 metabolism in tumor cells, NMR monitoring, 177, 449 molecular species analysis, 187, 195 reversed-phase HPLC, 172, 563 removal from plant extracts, 182, 192 vesicles ionic currents, estimation, 172, 341 preparation, 189, 374 turbidity measurement, 189, 380 Phosphomannosyl receptors assay, 179, 305 properties, 179, 309 purification from bovine testis, 179, 306 Phosphonium compounds quaternary, in determination of yeast membrane potential, 174, 611 Phosphoproteins effects of substrates and other ligands, 177, 277 liquid secondary ion MS analysis, 193, 480 metal ion binding, analysis, 177, 279 nuclear, detection and purification, 181, 29 pH titrations, 177, 274 3~p NMR studies, 177, 263 yeast, radiolabeling, 194, 423 Phosphoenolpyruvate carboxylase acetyl-CoA-independent, assay in crude extracts of methylotrophs, 188, 363 Phosphorescence in measurement of rotational and translational diffusion in membranes, 172, 471 Tb3+, in determination of biomembrane surface potential, 171, 372 Phosphorus in assay for platelet endogenous inositol phospholipids, 169, 426 Na+/P~ coupling ratio in inside-out vesicles, measurement, 173, 378

193 Phosphorylation cAMP-dependent, proteins in intact parietal cells, 191, 654 -dephosphorylation ratio, peptides, estimation, 193, 489 EGF receptor effect of ganglioside GM3, 179, 525 inhibition by IysO-GM3,179, 531 stimulation by de-N-acetyl-GM3,179, 531 mutagenic primers, 185,604 oxidative, see Oxidative phosphorylation proteins overview, 182, 642 in platelets, assay with membranepermeable diacylglycerol, 169, 434 sodium pump active site with inorganic phosphate, assay, 173, 688 tyrosine in angiotensin by pp60~', 193, 485 Phosphoserine identification by liquid secondary ion MS and automated sequencing, 193, 492 Photoaffinity labeling membranes, in identification of bile transport protein, 174, 28 Photoaffinity labels amiloride analogs, 191, 752 azides, 191, 767 Photobiotin acetate preparation and applications, 184, 588 Photobleaching fluorophores, minimization, 194, 576 Photochemical labeling bacteriorhodopsin, 172, 666 glycophorin A, 172, 662 hydrophobic, membranes applications, 172, 658 experimental aspects, 172, 653 integral membrane proteins, 172, 630 light-harvesting polypeptide B870-a from Rhodospirillum rubrum, 172, 667 membrane apolar phase, 172, 628 membrane-embedded protein domains, 172, 630 membrane lipid core in intact human erythrocytes, 172, 659 reagent requirements, 172, 633

Photon correlation spectroscopy proton-translocating portion F0 of FtF0ATP synthase, 172, 663 reagents for design and synthesis, 172, 648 distribution analysis, 172, 656 photoactivation, 172, 655 structure and application, 172, 672 Photochemistry caged nucleotides, 172, 298 in lipid bilayers, 172, 646 in organic solutions, 172, 646 Photodiode arrays in mass spectrometry, 193, 79 Photography in electron microscopy for RNA mapping, 180, 254 in immunoelectron microscopy of idiotype-anti-idiotype complexes, 178, 135 in in situ hybridization histochemistry with markers of neuronal connectivity, 168, 789 in 2D electrophoresis of proteins. 170. 137 Photolysis radical generation, principles, 186, 346 Photometry carbohydrate receptor ligand uptake by isolated cells, 179, 119 enzymes, substrates, and pathways in isolated nephron segments. 191. 337 low-angle light scattering, see Low-angle light-scattering photometry in measurement of endosomal and lysosomal pH, 173. 756 platelet aggregation, 169, 117 whole-cell, in measurement of cytoplasmic free Ca 2., 173, 765 Photomicroscopy yeast, 194, 572 Photomultiplier in measurement of fluorescence, 192. 43 Photon correlation spectroscopy in determination of vesicle size distribution, 172, 330 polydisperse samples, data evaluation, 172, 378

Photooxidation Photooxidation generation of oxygen radicals in vitro methods, 186, 637 in vioo methods, 186, 641 sensitized 2'-deoxycytidine, mechanism, 186, 510 thymidine, mechanism, 186, 506 tocopherols, 186, 200 Photoproteins assay, 172, 167 calcium contamination, control, 172, 168 calcium-regulated, as intracellular Caz+ indicators, 172, 164 desalting, 172, 170 introduction into cells, 172, 171 light signals, detection, 172, 181 lyophilization, 172, 170 sources, 172, 169 storage, 172, 170 Photoreceptors interpbotoreceptor matrix, see Interphotoreceptor matrix Photosensitization production of oxygen radicals, 186, 635 Phthalate dioxygenase assay, 188, 62 properties, 188, 66 purification from Pseudomonas cepacia, 188, 63 Phthalate dioxygenase reductase assay, 188, 62 properties, 188, 66 purification from Pseudomonas cepacia, 188, 65 Phycobiliprotein -avidin conjugates, preparation, 184, 188 -biotin conjugates, preparation, 184, 188 fluorescence, in assay for reactive oxygen species and retinoid antioxidant activity, 190, 277 Phycoerythrin -avidin conjugates, preparation, 184, 193 -biotin conjugates, preparation, 184, 193 fluorescence, in assay for reactive oxygen species, 186, 161 sources and preparation, 186, 162 Phylogenetic analysis, see also Evolution aspartic proteinases, 183, 682 cytochromes c, 183, 685 globins, 183, 688

194 molecular, statistical tests, 183, 645 nucleotide sequence divergence maximum likelihood methods, 183, 584 statistical estimation, 183, 531 protein sequences, tree construction, 183, 375 reconstruction and sequence alignment, unified approach, 183, 626 RNA secondary structure complementary sequences, 180, 234 hairpin structures, 180, 235 pseudoknots, 180, 300 sequence alignment, 180, 233 structure prediction from sequence, 180, 238 three-dimensional protein structures, 183, 670 tree determination with dynamically weighted parsimony method, 183, 615 visual pigment proteins, 183, 108 Physalaemin antibodies, selection for studies of physalaemin-related molecules, 168, 448 related molecules radioimmunoassay, 168, 448 recognition, purification, and structure, 168, 444 Physical maps Saccharomyces cerevisiae, 194, 827 Phytohemagglutin E4, -agarose affinity column, in chromatography of oligosaccharides, 179, 46 Pichia pastoris

purification of benzylamine/putrescine oxidase, 188, 429 Pisum sativum argenteum, see Pea Pithing shark rectal gland, 192, 762 Pituitary anterior, see Anterior pituitary GH3 cells cracked characterization, 168, 229 prolactin secretion activated by Ca2+, 168, 231 permeabilization to macromolecular probes, 168, 225

195 transient, in analysis of inositol lipid metabolism, 168, 218 human, perifusion in vitro, 168, 206 rat cell culture for in vitro bioassay of inhibin, 168, 602 preparation and phorbol ester pretreatment, 168, 288 protein kinase C-depleted, characterization, 168, 291 membranes, purification of gonadotropin-releasing hormone receptors, 184, 288 stimulus-response coupling, role of protein kinase C, 168, 287 surgical manipulation, 168,236 PK 11195 binding to isolated nephron segments, microassay, 191, 315 Placenta human, purification of cellular retinoic acid-binding protein, 189, 330 retinoid transfer, pharmacokinetics during organogenesis, correlation with teratogenicity, 190,437 Plants, see also specific plants cells breakage, 182, 183 components, preparation, 182, 175 elastic coefficients, 174, 205 implantation of pH microelectrode, 174, 335 passive permeability measurements, 174, 252, 262 preparation for plasmolysis, 174, 239 water relations dynamic relations, 174, 186 overview, 174, 191 pressure probe technique, 174, 338 water and solute flow, direct coupling, 174, 199 chloroplast extract preparation, 182, 186 extracts gel filtration posttreatment, 182, 193 nucleic acid and phospholipid removal, 182, 192 preparation, 182, 174 protein determination, 182, 182

Plasma frozen hydrated bulk specimens cryofracturing, 174, 270 quantitative X-ray microanalysis, 174, 274 growth coupling with water transport, 174, 211 limitation by solute relations, 174, 213 mechanical properties, coupling to water transport, 174, 204 phloem transport, analysis, 174, 288 thylakoid extract preparation, 182, 191 tissues elastic coefficients, 174, 205 preparation for plasmolysis, 174, 239 slicing, in isolation of vacuoles, 174, 554 water relations, 174, 191 water transport, 174, 189 water flow coupling to other processes, overview, 174, 183 theory, 174, 184 xylem transport, analysis, 174, 277 Plasma, see also Blood, Serum aromatic retinoids, HPLC, 189, 167 a- and fl-carotenes and other nutrients, simultaneous HPLC, 189, 155 catecholamines, assay, 168,435 citrated platelet-rich effects of aggregating agents, 169, 131 platelet aggregation in, effects of pH and divalent cations, 169, 120 preparation and characteristics, 169, 120 l l-dehydrothromboxane B2, assay, 187, 39 extracellular superoxide dismutase, automated assay, 186, 232 flow rate, in assessment of glomerular filtration, 191, 113, 125 glutathione and glutathione disulfide, analysis: assay of oxidant stress in vivo, 186, 682 high-molecular-weight kininogen antisera antibodies, preparation and characterization, 169, 280 coagulant assay, 169, 278 competitive ELISA, 169, 281 radiolabeling, 169, 279

Plasma histidine-rich glycoproteins immunoblot analysis, 169, 273 isolation and purification, 169, 269 [3H]retinol-labeled, preparation, 190, 305 human isolation of platelets by albumin gradient and gel filtration, 169, 11 centrifugation and washing, 169, 3 lipid hydroperoxides, HPLC-isoluminal chemiluminescence assay, 186, 375 phosphatidylcholine hydroperoxides, HPLC assay, 186, 159 platelet-rich labeling of platelets in, 169, 457 purification of platelet-derived growth factor, 169, 213,219 purification of high-molecular-weight kininogen, 169, 279 platelet-activating factor acetylhydrolase, 187, 352 retinol-bindingprotein, 189, 199 isotretinoin, HPLC, 189, 167 lycopene and other nutrients, simultaneous HPLC, 189, 155 purification of retinol-bindingprotein, 189, 2O3 rabbit, platelet-rich, labeling of platelets in, 169, 457 rat, purification of retinol-bindingprotein, 189, 195 retinoids, HPLC with automated column switching, 189, 146 retinol and other nutrients, simultaneous HPLC, 189, 155 tocopherols and other nutrients, simultaneous HPLC, 189, 155 Plasmalemma algal active transport studies, 174, 374 passive uniport, analysis, 174, 376 giant algal cell, permeabilization, 174, 440 Plasma membranes apical and basolateral, separation, 172, 18 basolateral isolation, 192, 396

196 vesicles for analysis of hepatic transport, preparation, 192, 535 canalicular, vesicles for analysis of hepatic transport, preparation, 192, 535 Chara cell isotopic fluxes, 174, 450 net fluxes, 174, 459 functional analysis, 172, 32 fusion-mediated implantation of exogenous proteins, 171, 846 hepatic isolation of bilitranslocase, 174, 50 isolation from rat, 182, 227 hepatocyte alanine transport, 174, 31 apical and basolateral vesicles protein analysis, 191, 836 resolution, 191, 831 vesiculation, 191, 829 biogenesis, 191, 825 sheets characterization, 191, 828 isolation from rat, 191, 826 sinusoidal, identification of bile acid transport protein, 174, 25 transport of glutathione species, 173, 523 isolation from polar cells and tissues, 172, 18 strategy, methods, and markers, 172, 3 markers alkaline phosphodiesterase I, assay, 182, 218 antigenic, enzymatic, and functional, 172, 15 Neurospora crassa, everted vesicles, isolation and transport function, 174, 667 permeability to normally impermeant compounds, alterations, 171, 858 permeabilization by high-voltage pulses, 171, 822 pore-forming chemicals, 171, 827 platelet, Na gradient, effect on calcium transport, 169, 383 proteins immunofluorescent detection, 194, 599 localization, 184, 637

197 purification, assessment, 172, 14 receptor purification, 184, 634 removal, 171, 819 Rhodotorula effect of N,N'-dicyclohexylcarbodiimide, 174, 647 electrochemical proton gradient, determination, 174, 632 fractions, isolation with cationic silica microbeads, 174, 648 permeability to H ÷ and K +, 174, 646 synaptosomal, Ca 2÷ cycling across, assay, 174, 8 tocopherol and tocopheryl ester incorporation and hydrolysis, 186, 355 vesicles contralumenal isolation from kidney, 191, 450 purity, 191,460 isolation from medullary thick ascending limb, 191, 461 renal inner medulla, 191, 465 lumenal isolation from kidney, 191,450 purity, 191, 460 yeast, enzyme marker assay, 194, 658 Plasmids ADH2 promoter, construction, 185, 309 in construction of genomic banks for yeast transformation, 194, 199 construction by homologous recombination in vivo, 185, 294 DNA transmission to yeast by conjugation with bacteria, 194, 187 dual, repressible system for Bacillus subtilis, 185, 208 with Escherichia coli trp promoter, construction, 185, 56 GAPDH expression cassette, construction, 185, 344 instability in Bacillus subtilis, 185, 200 toxic gene-related, prevention, 185, 80 integrative transformation of Bacillus subtilis, 185, 225 yeast, 194, 281 vectors for protein production by secretion from yeast, 194, 494

Plasmids manipulation, 185, 35 in manipulation of yeast genome, 185. 280 with multicopy insert in head-to-tail orientation, construction, 182, 522 pACYC177-1aciO, for transcription optimization in Escherichia coli, 182, 96 pAdD26SVpA3, for gene coamplification in mammalian cells, 185, 560 pASKI9, expression vector for functional Fv and Fab fragments in Escherichia coli, 178, 507 pASK22, expression vector for functional Fv and Fab fragments in Escherichia coli, 178, 507 pATH, high-expression vector for construction of trpE fusion genes, 194, 477 pBLI, inducible secretion vectors for Bacillus subtilis, construction. 185. 213 pDH51, for protoplast transformation. 181. 154 pET, in T7 RNA polymerase-directed expression of cloned genes. 185, 65 pG-l, in constitutive gene expression in yeast, 194, 391,397 pG-2, in constitutive gene expression in yeast, 194, 391,397 pG-3, in constitutive gene expression in yeast, 194, 391,397 pGAL-HO, delivery of double-strand breaks, 194. 140 pGTy, in mutagenesis in Saccharomyces cerevisiae, 194, 343 pGV403, inserted DNA, nucleosome core particle assembly on, 170,496 pGX2134, apr[BamP]-based, description. 185, 215 priSe, bacterial hosts bearing, production of T4 lysozyme from, 177.72 pHZI8, construction, 181, 123 pJDB219, based vectors for propagation of cloned genes in yeast, 185, 252 pL. for transcription optimization in Escherichia coli, 182, 94 pLyE, in T7 RNA polymerase-directed gene expression in Escherichia coli, 185, 63

Plasmids pLyS, in T7 RNA polymerase-directed gene expression in Escherichia coli, 185, 63 pMA91, PGK promoter-based, as highefficiency yeast expression vectors, 185, 329 pMON5743 construction, 185, 116 description, 185, 119 enhancement of foreign gene translation in Escherichia coli, 185, 115 pREP9 foreign gene introduction into, 185, 207 inducible secretion vectors for Bacillus subtilis, construction, 185, 213 vector for inducible gene expression in Bacillus subtilis, 185, 205 pRIT20 and pRIT30 series design and construction, 185, 152 fusion proteins from, expression and purification, 185, 155 for staphylococcal protein A fusions, selection, 185, 153 pSV2, in production of chimeric antibodies, 178, 462 pSV2gP', for expression of recombinant Igs with effector functions, construction, 178, 522 pT7, for transcription optimization in Escherichia coli, 182, 96 pUCI8 and pUC19 fusion vectors for translation optimization, 182, 104 with lac promoter, for transcription optimization in Escherichia coli, 182, 96 p2UG, vector for inducible gene expression in yeast, 194, 394 for recombinant protein production in mammalian cells, selection, 185, 567 rnpA-carrying, cells with, purification of C5 protein, 181, 573 rnpB-carrying, cells with, preparation of M1 RNA, 181,578 Schizosaccharomyces pombe description, 194, 806, 810 integration into genome, 194, 814 recovery, 194, 814 stability, testing, 194, 813 transformation methods with, 194, 808

198 for secretion of heterologous gene products, components, 185, 422 shuffling, in generation of mutant yeast from cloned genes, 194, 302 shuttle vectors for expression of cloned genes in yeast, 194, 388 recovery from yeast into Escherichia coli, 194, 319 shuttling, yeast DNA miniprep for, 185, 293 td , in analysis of RNA splicing in Escherichia coli, 181, 522 templates for small RNA synthesis, 180, 54 transfection of Sf9 cells, 182, 121 transient expression in COS-I cells, 185, 507 for transient transfection of HeLa cells in analysis of splicing, 181, 45 for trpE fusions, construction, 185, 163 2-/zm applications, 185, 250, 268 composition, 185, 237 copy levels, 185, 249 examples, 1.85, 238, 261,270 for high copy propagation of cloned genes in yeast, 185, 252 in propagation and expression of cloned genes in yeast, 185,234 stability, 185, 248 vectors in Escherichia coil, design and construction, 185, 14 yeast ADH2 promoter-containing, construction, 185, 311 allowing regulated secretion of heterologous proteins, construction, 185, 312 bacterial transformation, 194, 323 centromere, in genomic manipulation, 185, 290 chromatin, isolation, 170, 26 integrating: applications of homologous recombination, 185, 285 preparation, 194, 322 shuffling in vitro, 194, 302 2-/zm circle, in propagation and expression of cloned genes, 185, 234

199 vectors for constitutive and inducible gene expression, 194, 389 expression of cloned genes, 194, 373 heterologous gene expression, 185, 231 YCp vectors, in genomic manipulation, 185, 290 YEp and YCp vectors borne genomic banks acquisition by mail, 194, 214 applications, 194, 224 construction de nooo, 194, 216 incompleteness, 194, 229 for construction of genomic banks, selection, 194, 209 properties, 194, 199 YIp vectors, in genomic manipulation, 185, 285 Plasminogen activator differentiation marker for embryonal carcinoma cells, 189, 344 human urokinase-type, secretion from yeast, screening, 194, 502 tissue-type in oocytes assay, 168, 424 neuroendocrine regulation, 168, 422 recombinant antifibrin antibody with, production and analysis, 178, 515 Plasmodium falciparum

isolation by free-flow electrophoresis, 171, 521 Plasmolysis alteration of protoplast, 174, 229 applications, 174, 241 experimental procedures, 174, 233 form and time, influencing factors, 174, 229 in measurement of passive permeability, 174, 250 Plasmolyticurn selection and preparation, 174, 233 Plastic cell adhesion, 190, 82 Plate detectors in mass spectrometry, 193, 62 Platelet-activating factor antagonists WEB2086 and WEB2170, properties and applications, 187, 455

Piatelets binding sites, interactions with ginkgolides, 187, 433 bioassay by [3H]serotonin release, 187, 130 rabbit platelet aggregation, 187, 125 GC/MS assay, 187, 134 GC-NCI-MS assay, 187, 142 half-life determination, 187, 347 hybrid sector-quadrupole MS analysis, 193, 199 production by endothelial cells, 187, 527, 533 release and effects in isolated coronaryperfused mammalian heart, 187, 610 TLC, 187, 152 Platelet-activating factor acetylhydrolase assay, 187, 345 properties, 187, 348 purification, 187, 352 substrate specificity, 187, 355 Platelet-activating factor receptors antagonists, screening in vitro, 187, 447 Platelet-derived growth factor cell culture assays, 169, 214 characterization, 169, 215 induced chemotaxis of inflammatory cells and fibroblasts assays, 169, 236 checkerboard analysis, 169, 241 localization in platelet a granules, 169. 212 purification from human platelets, 169, 215 outdated platelet-rich plasma, 169, 219 sources of purification and characterization, 169, 213 Platelet-derived growth factor receptors associated tyrosine kinase, effects of gangliosides GM3 and GMI, 179, 523 Platelet factor 4 low-affinity, purification from platelets, 169, 225 Platelet heparitinase assay, 169, 343 properties, 169, 350 purification, 169, 346 Platelets adhesion definition, 169, 39 to polymeric surfaces, ex vivo measurement, 169, 91

Platelets adhesive macromolecules, dark-field electron microscopy, 169, 326 agglutination/aggregation tests, 169, 149 aggregation in analysis of platelet histidine-rich glycoprotein, 169, 274 assay, 187, 447 congenital abnormalities, 169, 133 general characteristics, 169, 118 measurement in vitro by microscopic and electronic particle counting, 169, 134 PAF-induced, in analysis of WEB 2086 and 2170, 187, 458 photometric measurement, 169, 117 a2-antiplasmin, radioimmunoassay, 169, 298 arachidonic acid metabolism, 187, 582 aspirin treatment, 187, 588 classification, 169, 38 collection and processing, 187, 587 cytoplasmic Ca2+ levels, measurement, 169, 386 deposition data, 169, 104 definition, 169, 40 eicosanoid interactions, 187, 585 fixation, 169, 149 fixed in bioassay for von Willebrand factor, 169, 155 yon Willebrand factor testing with, applications, 169, 160 in flowing blood, interactions with vessel wall components, 169, 37 function, effects of lymphocytes, 187, 584 granule proteins, immunocytochemical localization, 169, 301 a granules, localization of plateletderived growth factor, 169, 212 high-molecular-weight kininogen coagulant assay, 169, 278 expression on activated platelet surface, 169, 290 function, 169, 295 [3H]serotonin release, in bioassay of platelet-'activating factor, 187, 130 human applications of lipid assays, 169, 427

200 arachidonic acid metabolism, 187, 589 binding of thromboxane A2/prostaglandin H2 receptor antagonists, 187, 410 calpain, natural substrates, 169, 454 comparison with porcine platelets, 169, 25 degradation of extracellular matrix components, 169, 85 function, role of surface-bound Ca2+, 169, 369 isolation from plasma by albumin gradient and gel filtration, 169, 11 centrifugation and washing, 169, 3 labeling in platelet-rich plasma, 169, 457 membranes crude preparations, 187, 399 solubilized, preparation, 187, 399 preparation, 169, 356 purification of calpains and calpastatin, 169, 445 platelet-derived growth factor, 169, 215 thrombospondin, 169, 256 sample collection for thrombospondin radioimmunoassay, 169, 262 stimulated collection and processing, 169, 416 endogenous inositol phospholipids, analyses, 169, 419 surface-bound Ca2÷, assay, 169, 365 surface membrane Cae+-binding proteins in, identification and characterization, 169, 365 interaction with extracellular Ca2~, 169, 355 thrombospondin secretion, 169, 251 unstimulated, surface-bound Ca2+, assay, 169, 356 washed binding of ~2q-labeledligands, 187, 401 preparation, 187, 398 suspensions, preparation and labeling, 169, 459 "lln-labeled, in vivo turnover and organ distribution, 169, 172

201 intact, calcium transport, analysis, 169, 371 integrity during and after loading with aequorin, 169, 391 Quin 2, 169, 404 interactions with artificial surfaces in vitro, 169, 104 endothelial monolayers, 169, 71 subendothelial extracellular matrix, 169, 76 isolation from laboratory animals, 169, 27 for yon Willebrand secretion studies, 169, 245 labeling with radioactive arachidonate, 187, 590 -lymphocyte interactions in prostacyclin biosynthesis, 187, 578 membranes, binding of thromboxane AJ prostaglandin H2 receptor antagonists, 187, 412 partial purification of protein kinase C, 169, 439 perfused hearts, eicosanoid and plateletactivating factor studies, 187, 614 plasma membrane Na gradient, effect on Ca transport, 169, 383 platelet-derived growth factor cell culture assays, 169, 214 characterization, 169, 215 localization in a granules, 169, 212 purification sources, 169, 213 porcine comparison with human platelets, 169, 25 isolation from blood, 169, 22 preparation, 169, 284; 187, 578 preparations, effects of anticoagulants, 169, 147 purification of heparitinase, 169, 346 high-molecular-weight kininogen, 169, 279 histidine-rich glycoprotein, 169, 269 PADGEM protein, 169, 315 platelet basic protein and low-affinity platelet factor 4, 169, 225 rabbit aggregation, in bioassay of pafacether, 187, 125

Platelets in vivo studies, application of prelabel-

ing methods, 169, 466 labeled, preparation for transfusion. 169, 468 labeling in platelet-rich plasma. 169. 457 membranes, ginkgolide interactions with PAF binding sites, 187. 435. 438 protein phosphorylation, assay, 169. 434 secretion of /3-N-acetylglucosaminidase, 169, 435 serotonin, 169, 435 serotonin release in blood, assay, 169, 470 washed, suspensions, preparation and labeling, 169, 458 radiolabeling, 169, 94 responses to aggregating agents, 169. 131 secreted yon Willebrand factor, multimeric structure, 169, 249 secretion of adenine nucleotides, assay. 169, 195 high-molecular-weight kininogen, 169. 286 histidine-rich glycoprotein, 169, 272 lysosomal acid glycosidases, assay. 169, 336 serotonin assays, 169, 196, 435 effects of diacylglycerol and Ca 2~ ionophore, 169, 436 fluorimetric assay, 169, 208 radioactive assay, 169, 206 yon Willebrand factor assay by radioquantitative electrophoresis, 169, 248 release experiments, 169, 247 secretory pathways, overview, 169, 191 semiautomated counters, in particle analyzers, 187, 447 separation from plasma proteins, 169. 33 -surface interactions analysis artery segments for, processing. 169, 62 perfusates for, preparation. 169, 50 perfusion chambers for. 169, 43

Platelets effects of physical and chemical factors, 169, 41 evaluation in absence of fibrin formation, 169, 59 in presence of fibrin formation, 169, 69 test surfaces, preparation, 169, 47 suspensions radiolabeled, experimental use, 187, 591 washed, preparation, 187, 588 thrombin-stimulated releasate, immunoblot analysis, 169, 273 fl-thromboglobulin antigens, identification and distribution, 169, 227 thrombus formation, volume, height, and growth, definition, 169, 41 volume and number determination, 169, 164 washing to obtain von Willebrand factor, 169, 246 for yon Willebrand factor release studies, 169, 247 Plating keratinocytes on fibroblast feeder layer, 19tl, 22 replica, see Replica plating cis-Platinum, see Cisplatin PLFASTA program evaluation of local sequence similarity, 183, 88 Polar cells isolation of plasma membranes, 172, 18 Polarization fluorescence, in analysis of nucleosomes, 170, 202 Polarographic assay gentisate 1,2-dioxygenase, 188, 102 methane monooxygenase, 188, 192 methylamine oxidase, 188, 227 protocatechuate 2,3-dioxygenase, 188, 95 protocatechuate 3,4-dioxygenase, 188, 83 protocatechuate 4,5-dioxygenase, 188, 89 Polidexide in ion-exchange chromatography of RNA, 1811, 26 Poliovirus monoclonal anti-idiotypic antibody vaccines, 178, 375

202 Poly-N-acetyllactosamine glycans cell surface, specific labeling, 179, 270 Poly-N-acetyllactosaminoglycans biosynthesis, 179, 359 Polyacrylamide beads -antibody, in solid-phase immunoassay of human IgA, 182, 709 Polyacrylamide gels calibration with marker proteins for electrophoresis, 182, 439 casting, 182, 431 covalent immobilization of carbohydrates on, 179, 542 formation and properties, 182, 427, 462 granulated, for preparative isoelectric focusing, 182, 475 native, preparation, 180, 448 RNA in, extraction, 180, 429 Polyadenylate in analysis of nucleotide linkage of RNA ligase product, 181, 493 binding proteins identification, 181, 333 manipulation in Saccharomyces cerevisiae, 181, 347 purification and characterization, 181, 332 synthesis requirements, 181, 165 Polyadenylate polymerase antibodies, effect on polyadenylation in vitro, 181, 67 assay, 181, 167 properties, 181, 164 purification, 181, 162 Polyadenylation detection, 181, 55 effect of specific antibodies in vitro, 181, 67 mammalian extracts active in, preparation, 181, 49 mediating factors, identification, 181, 67 precursor mRNA, in expression of cDNA genes in mammalian cells, 185, 502 reaction conditions, 181, 54 RNA substrates, preparation, lgl, 52 specific complexes, detection, 181, 63 specific events, assays, 181, 54 Polyaminocarboxylates chelation of metals, 186, 116

203 -metal complexes, reactions with free radicals, 186, 117 reaction with OH/O radicals, 186, 117 Poly(A) polymerase, see Polyadenylate polymerase Polybrene, see Hexadimethrine bromide Polybuffer separation from protein, 182, 389 Polyclonal antibodies advantages and disadvantages, 182, 664 antihistone, preparation, 170,256 anti-idiotypic to antimorphine antibodies assay, 178, 228 production, 178, 227 purification and properties, 178, 234 antiopioid receptor, production and characterization, 178, 243 generation with complementary peptides, 178, 66 anti-PADGEM, preparation, 169, 313 arachidonate 5-1ipoxygenase, preparation, 187, 493 detection methods, comparison, 182, 668 idiotypic, generation with complementary peptides, 178, 66 preparation animal selection, 182, 666 antigen purification and preparation, 182, 663 from rabbit serum, 182, 665 purified, biotinylation, 184, 471 Polycyclic aromatic hydrocarbons biodegradation by M y c o b a c t e r i u m , 188, 149 Poly(l ,Nr-ethenoadenylic acid) fluorescence, in measurement of polyadenylate-bindingprotein dissociation rates, 181, 347 Polyethylene glycol -dextran-water system for partition of cells and organelles, 171, 532 precipitation of proteins, 182, 301 reference ion masses, 193, 872 removal from protein, 182, 305 Polyethyleneimine precipitation of nucleic acids and phospholipids in plant extracts, 182, 192 Polygalacturonase in digestion of polygalacturonic acid, 179, 567

Polypeptides Polygalacturonic acid enzymatic digestion, 179, 567 oligomer isolation, 179, 568 Poly(glutamic acid) in reconstitution of chromatin from purified components, 170, 585 Polymerase chain reaction cDNA amplification for nuclease digestion method of intron mapping, 180. 333 in cloning of nuclear retinoic acid receptors, 189, 256 in hybrid receptor and protein synthesis, 189, 261 Polymerases types 1, II, and II1, encoding genes, identification, 180,497 Polymerization B-cell peptide with T-cell peptide, 178. 664 Polymeric shunts merits in ex vivo studies, 169, 98 platelet and thrombus deposition data, 169, 104 preparation and protein preadsorption. 169, 95 surgical placement, 169, 95 Polymers coated surfaces, platelet adhesion, ex vivo assay, 169, 91 Polymin P in immunoprecipitation of RNA polymerase II, 194, 517 Polymyxin B rotating-frame NOE, 176, 202 Polynucleosomes preparation, 170, 185 Polynucleotide kinase in 5'-end labeling of RNA, 180, 194 in labeling of RNA digests for fingerprinting, 180, 135 T4, in 5' labeling of DNA primers, 180. 357 Polypeptides B870-c~ from Rhodospirillum rubrurn, photochemical labeling, 172, 667 components of Na+-D-glucose cotransporter, partial purification, 191, 603 cotranslational N-terminal processing, 185, 398

Polypeptides domains crosslinked to DNA, mapping and identification, 170, 403 hybrid, yeast, purification for antigenic use, 194, 488 membranous sites, ion interactions, NMR studies, 171, 286 small nuclear ribonucleoprotein applications, 181, 263 isolation, 181, 257 transit through yeast secretory pathway, 194, 662 in tRNA splicing endonuclease, analysis, 181,480 Polyproteins HIV-1 gag, k-tuple frequency analysis, 183, 244 Polyribonucleotide synthase (ATP) -AMP complex, assay and characterization, 181,497 in 3'-end labeling of RNA, 180, 196 wheat germ assay, 181, 488 associated activities assay, 181, 488 purification and characterization, 181,480 product nucleotide linkage, analysis, 181, 492 purification and characterization, 181, 480 Polyribosomes discovery by sucrose gradient analysis, 170, 97 extraction of rRNA, 181, 362 preparation, 181, 362 salt extraction, 181, 205 Polysaccharides acidic, separated by gel electrophoresis, immunoblotting, and immunobinding, 179, 95 biotinylation, 184, 539 capsular, bacterial, gel electrophoresis, 179, 104 conformation, 2D ~H NMR studies, 179, 153 linkage analysis by reductive cleavage method, 193, 573 Polysomes, see Polyribosomes Polystyrene balls, immobilization of proteins, 184, 488

204 Poly(U)-Sepharose in affinity chromatography of RNA, 180, 36 Populations human, vitamin A status, biochemical and histological assessment, 190, 242 Pores kinetics, comparison with simple carrier, 173, 152 Porins outer membrane, OmpF, secretion from Escherichia coli, 182, 136 Position-resolved arrays in mass spectrometry, 193, 84 Postacceleration detectors in mass spectrometry, 193, 72 Potassium activation site of H+,K+-ATPase, reversible antagonists, interaction with ATPase, 191, 737 active transport in turtle colon, 192, 736 conductance in intestinal secretory cell membranes characterization, 192, 309 pharmacological properties, 192, 316 turtle colon basolateral membrane analysis, 192, 740 role of intracellular Ca 2÷, 192, 742 fluxes through sodium pump, 173, 92 - H ÷ symport in Neurospora, 174, 654 electrophysiological analysis, 174, 659 ion flux analysis, 174, 662 properties, 174, 664 intracellular, assay in intestinal epithelial cells, 192, 337 low levels, effect on number of sodium pump molecules in cultured heart cells, 173, 651 Na÷/K+/C1 - cotransport system analysis in intestinal cell lines, 192, 371 identification and purification, 191, 786 Na+-K + pump in muscle fibers, stoichiometric ratio, 173, 707 rate measurements with radioactive tracers, 173, 700 stoichiometry and coupling in erythrocyte inside-out vesicles, 173, 377 net accumulation in muscle, 173, 697

205 net flux in lepidopteran midgut, agreement with Is¢, 192,607 outflow from yeast, assay in presence of glycine, 174, 625 permeability of R h o d o t o r u l a plasma membrane, 174, 646 in sweat gland tissue, assay, 192, 596 transport in kidney, effects of furosemide-related diuretics, 191, 785 Potassium channels Ba2+-sensitive, ion fluxes in membranes from renal outer medulla, 172, 162 Ca2*-activated, in human red cells, patch-clamp studies, 173, 119 Ca2*-dependent, in one-step inside-out vesicles, assay, 173, 374 characterization with amiloride analogs, 191, 746 electrically gated, detection in R h o d o t o r u l a , 174, 647 in intestinal cell lines, analysis, 192, 372 Potassium chloride precipitation of dodecyl sulfate-protein complexes, 170, 395 Potato purification of arachidonate 5-1ipoxygenase, 187, 296 tubers, purification of arachidonate 5lipoxygenase, 187, 268 Potentiometry optical, in measurement of membrane potential of cells, organelles, and vesicles, 172, 102 signal optimization, 172, 112 Precipitation alum, antibody coupled to carrier protein, 178, 46 cetylpyridinium chloride-induced, in platelet heparitinase assays, 169, 343 co-, s e e Coprecipitation in detergent removal from membrane proteins, 182, 279 differential in isolation of brush border membranes, 191, 451 in preparation of plasma membranes, 172, 11 divalent cation, in isolation of brush border membranes, 192, 353

Pressure dodecyl sulfate-protein complexes, 170. 395 heat denaturative, in enzyme purification, 182, 298 immuno-, s e e lmmunoprecipitation mannoprotein with Cetavlon, 185,456 Na ~ cotransport proteins, 191, 587 nucleic acids from plant extracts, 182, 192 protein samples, 186, 466 pH denaturative, in enzyme purification, 182,298 phospholipids from plant extracts, 182. 192 in protein concentration and solute removal, 182, 78 proteins with acetone and acid, 182, 592 bulk methods, 182, 292 fractionation with ammonium sulfate, 182, 293 heat denaturative methods, 182, 298 historical overview, 182, 285 with NaCI and other univalent-univalent salts, 182, 298 with organic solvents, 182, 296 pH denaturative methods, 182, 298 with polyethylene glycol, 182, 301 theory, 182, 287 with trichloroacetic acid and deoxycholate, 182, 593 selective, biotin-labeled antigens or monoclonal antibodies, 184, 518 uncrosslinked proteins from D N A protein sample, 170, 393 Preproenkephalin transcripts, detection in rat brain and cultured adrenal medulla, 168, 688 Presaturation in solvent suppression for NMR, 17@ 66 Preservation yeast strains, 194, 18 Pressure effects on amino acid transport in red cells, 173, 156 collision-induced dissociation, 193, 242, 253 hydraulic conductivity, 174, 197

Pressure hydraulic, transcapillary, in assessment of glomerular filtration, 191, 110, 123 negative, measurement, 174, 358 osmotic, see Osmotic pressure pressure probe measurement, constralnts, 174, 356 relaxation, analysis, 174, 342 root, probe method, 174, 218 Pressure probe method in determination of elastic modulus of cell wall, 174, 353 hydraulic conductivity, 174, 354 negative pressure, 174, 358 reflection coefficients, 174, 349 pressure clamp experiments, 174, 343, 362 pressure measurements, constraints, 174, 356 theory, 174, 339 volume measurements, constraints, 174, 356 Primer extension analysis indirect mapping of UV-induced crosslinks, 180, 438 mapping of RNA branches, 181, 184 oligodeoxynucleotide/RNase H digestion products, 181, 292 RNA, 180, 347 artifacts, 180, 367 hybridization, 1811, 361 interpretation, 180, 366 primer selection and preparation, 180, 352 products characterization, 180, 368 sequence determination, 180, 368 rRNA termini maturation in Escherichia coli, 181, 359 sensitivity, increase with continuous labeling, 180, 367 structural probing of native snRNA, 180, 221 yeast D N A sequencing and footprinting, 194, 550 yeast pre-mRNA sequence contribution to splicing efficiency, 181, 126, 141 Primers mutagenic, synthesis and phosphorylation, 185, 604

206 Procholecystokinin antisera, development, 168, 3 Prochymosin bovine, secretion from Escherichia coli, 182, 136 yeast, screening, 194, 498 Profile analysis sequence data, 183, 146 Profilometry optical, in analysis of effects of topical retinoids on photoaged skin, 190, 360 Prohormones processing, analysis with antibodyHPLC method, 168, 517 Proinsulin human, encoding gene, expression with two-cistron system, 185, 101 Prokaryotes extract preparation enzymatic lysis, 182, 147 mechanical lysis, 182, 151 nucleic acid removal, 182, 153 Mg2÷ transport, assay, 173, 546 proton electrochemical potential gradient in, 172, 63 subcellular isotope distribution, determination, 173, 553 Prolactin binding to membrane fractions assay, 178, 315 inhibiting anti-idiotypic antibodies, potency, 178, 316 secretion from anterior pituitary single cells, cell blot assay, 168, 327 Ca2+-activated, from cracked GH3 cells, 168, 231 superfusion and static cell culture assay methods, 168, 254 Prolactin receptors anti-idiotypic antibodies as probes, 178, 308 membrane-bound, preparation, 178, 313 Proline spin system identification, 177, 139 Prolyl endopeptidase interference with glutaminylpeptide cyclase assay, 168, 363

207 Pronase in dissociation of gastric mucosa into individual cells, 171, 446 in isolation of nonparenchymal rat liver cells, 190, 53 Propane metabolizing enzymes, cell-free assays, 188, 26 Propane oxygenase assay, 188, 28 Propionate-CoA ligase assay, 188, 31 Propionyl-CoA synthetase, s e e Propionate-CoA ligase Prostacyclin biosynthesis via platelet-lymphocyte interactions, 187, 578 circulating in rabbits, assay, 169, 462 production by endothelial monolayers, role of platelets, 169, 75 synthesis by human umbilical vein endothelial cells, 187, 539 Prostacyclin synthase detection in lymphocytes, 187, 583 Prostaglandin E~ -Triton X-100 complex, sedimentation coefficient and partial specific volume, 172, 703 Prostaglandin E2 production by parietal cells, effects of gastric secretagogues, 187, 513 synthesis by NIH-3T3 murine fibroblasts, 187, 541 Prostaglandin endoperoxide synthase, s e e Prostaglandin synthase 9a, 1l/3-Prostaglandin F2 stable isotope dilution MS assay, 187, 51 Prostaglandin H2 biosynthesis, 187, 247 platelet-derived, role in prostacyclin synthesis by lymphocytes, 187, 583 purification and analysis, 187, 245 Prostaglandin H synthase, s e e Prostaglandin synthase Prostaglandin 12, s e e Prostacyclin Prostaglandins biosynthesis-related enzymes, nomenclature, 187, 5 catabolism-related enzymes, nomenclature, 187, 5

Protein A effects on isolated perfused mammalian heart. 187, 613 parietal cell acid production, 187, 512 synthesis, determination with isolated glomeruli, 191, 135 Prostaglandin synthase apoenzyme assay, 187, 480 preparation and proteolysis, 187, 479 cyclooxygenase activity, activation, in assay of hydroperoxides, 186, 435 generation of glutathione thiyl radicals. 186, 324 murine cloning, 187, 469 primary structure, 187, 477 ovine cloning, 187, 469 isolation from seminal vesicles, 186. 432 primary structure, 187, 477 oxidation of reducing cofactors, 186. 286 oxygenation of arachidonic acid, 186, 285 proteolytic fragments, peptide mapping. 187, 484 trypsin-cleaved, reversed-phase HPLC, 187, 483 trypsin digestion, 187, 482 Protamine displacement, in isolation of histone dimers and tetramers, 170, 447 PROTEAN program protein solution structure determination from NMR data by heuristic refinement, 177, 222 Protease A Saccharomyces,

see Saccharomyces

aspartic proteinase Protease B yeast, s e e Yeast proteinase B Proteases, s e e Proteinases Protein A biotinylated, in ELISA of JE antibodies, 184, 497 component of soluble methane monooxygenase, purification from M e t h y l o c o c c u s c a p s u l a t u s Bath, 188, 184

Protein A in matrix for detection and purification of inhibin, 168, 605 staphylococcal, based gene fusion expression system, 185, 144 Proteinase inhibitors in buffers for solubilization of membrane proteins, 182, 259 exogenous, prevention of proteolysis during yeast extract preparation, 182, 170 prevention of proteolysis, 182, 87 type I, synthesis in tomato plants, induction by di- and triuronides, 179, 569 Proteinases deficient yeast, design, 185, 385 defective hosts, in enhancement of protein product stability, 182, 106 Lon, mutations minimizing proteolysis in Escherichia coli, 185, 120 membrane protein resistance to, analysis, 191, 852 purified, in assay of protein degradation, 186, 502 secreted by Bacillus subtilis, effects on protein stability, 185, 200 yeast, see Yeast proteinase B Protein B purification from Methylococcus capsulatus Bath, 188, 185 Protein C assay, 188, 183 purification from Methylococcus capsulatus Bath, 188, 186 Protein folding analysis with antibodies, 191, 851 intermediates, ~H NMR studies dynamics at equilibrium, 176, 462 folding-unfolding transitions at equilibrium, 176, 447 pathways, characterization by hydrogen exchange labeling, 176, 465 stability, 176, 451 recombinant proteins, 185, 187 recombinant tissue inhibitor of metalloproteases, 185, 192 secretory leukocyte protease inhibitor, 185, 190 solubilized protein aggregates from Escherichia coli, 182, 270

208 Protein-glutamine y-glutamyltransferase type I assay, 190, 43 down-regulation by retinoids, 190, 42 Protein Identification Resource applications, 183, 34 database acquisition, 183, 49 database type, 183, 32 entries fully annotated, 183, 38 minimal, 183, 42 history, 183, 36 as international collaboration, 183, 45 newsletters, acquisition, 183, 49 nomenclature, 183, 47 software, acquisition, 183, 49 superfamily-based organization, 183, 37 Protein kinase cAMP-dependent assay in gastric parietal cells, 191, 650 hormonal activation, detection in gastric parietal cells, 191,652 isozyme identification, 191, 651 characterization with amiloride and analogs, 191, 746 Protein kinase C activation, role of arachidonate metabolism, 191, 684 assay, 168, 289 depleted pituitary cells, characterization, 168, 291 effects of Gm and derivatives in A431 cells, 179, 536 extraction from pituitary cells, 168, 289 platelet assay, 169, 437 partial purification and properties, 169, 439 rat brain assay, 168, 348 subspecies resolution, 168, 347 role in arachidonate release from phospholipids, 191, 682 stimulus-response coupling in pituitary, 168, 287 Proteins acetylation, inhibition, 185, 406 ACTH-binding assay, 184, 267

209 biotinylation, 184, 264 isolation, 184, 262 properties, 184, 270 active sites, structure-function analysis with anti-idiotypic antibody, 178, 163 activity-based analysis, 182, 557 activity loss, evaluation, 182, 88 adenosine deaminase-binding, competitive idiotype-anti-idiotype enzyme immunoassay, 178, 171 adipocyte lipid-binding purification from human and mouse, 189, 363 retinoid binding, 189, 366 ADP-ribosylation, 182, 637 ADR1, overexpression, effects on A D H 2 promoter, 185, 315 adsorption to gold particles, 179, 114 affinity chromatography general methods, 182, 357 on immobilized reactive dyes, 182, 343 for protein concentration, 182, 83 aggregates in Escherichia coli inclusion bodies containing, washing, 182, 268 purification, 182, 273 refolding, 182, 270 solubilization, 182, 264 alanine transport, identification in hepatic cell membrane, 174, 38 aliphatic side chains, connectivities along, tracing, 176, 162 amino acid analysis, overview, 182, 587 amino acid sequences in prediction of B-cell determinants, 178, 586 sensitivity comparison, 183, 352 amino-terminal residue N-myristoylation, 182, 640 structure selection, 185, 404 ampholyte removal, 182, 389,477 analysis by collision-induced dissociation of multiply charged molecular ions, 193, 428 anion-exchange, isolation, reconstitution, and transmembrane orientation, 173, 410 anion transport, see Anion transporter

Proteins antigenic segments, identification by hydrophilicity plotting, 178~ 571 aqueous humor, levels: bioassay of arachidonic acid metabolites, 187, 381 AraC family, detection with DNA databases, 183, 121 aspartate residues /3-carboxylation, 182, 635 carboxyl methylatiom 182. 632 assays cluster tray method, 171, 138 colorimetric methods, 182, 57 glass cuvettes in, 182, 65 interfering substances, removal, 182. 67 overview, 182, 50 in presence of high lipid levels, 172. 393 protein standards, 182, 66 ultraviolet absorption methods, 182. 50 association with band 3 at cytoplasmic surface of erythrocyte membrane. 173, 513 -avidin conjugates, preparation, 184, 177 band 2.1, purification from red cell membranes, 173, 385 band 3 binding of stilbene disulfonates, fluorescent assay, 173, 510 circular dichroism spectroscopy, 173, 511 cyanogen bromide coupling, 173, 507 detection in Sendal virus envelopes, 171, 838 erythroid, mediated anion transport in mRNA-injected X e n o p u s oocytes. 173, 453 functional asymmetry, effects on substrate and inhibitor binding, 173,432 fusion-mediated implantation into Friend erythroleukemia cell plasma membranes, 171. 846 immobilization, 173, 506 in intact red cell, protein ligand association with, 173, 518 membrane domain, purification, 173, 508

Proteins monomers, matrix binding, 173, 506 oligomers crosslinking, 173, 512 matrix binding, 173, 506 protein associations at cytoplasmic surface of erythrocyte membrane, 173, 513 purification and characterization, 173, 494 solubilization, 173, 499 in solution native structure, assay, 173, 509 protein ligand association with, 173, 517 -Triton X-100, sedimentation coefficient and partial specific volume, 172, 700 band 4.1 purification from red cell membranes, 173, 385 radiolabeling, 173, 387 barrier, activity in yeast, assay, 194, 89 bile acid transport, identification in hepatocyte sinusoidal plasma membranes, 174, 25 binding of halide ion, quadrupolar NMR study, 177, 330 4-hydroxyalkenal, 186, 405 binding to RNA, protein sequestering assay, 181, 277 biotin-binding nonavidin, overview, 184, 93 overview, 184, 49 biotinylated affinity purification, 184, 53 detection, 184, 435 effect of spacer length, 184, 489 biotinylation via amino groups, 184, 139 with biotinyl N-hydroxysuccinimide ester, 184, 140 on blots, 184, 422 before blotting, 184, 418 via carboxyl groups, 184, 154 with p-diazobenzoylbiocytin, 184, 151 enzymatic, at carboxyl terminal, 184, 160 on exocytic transport vesicle or plasma membrane surfaces, 191, 822

210 with 3-(N-maleimidopropionyl)bioc ytin, 184, 153 on nitrocellulose replicas, 184, 434 for visualization of intracellular trafticking, 184, 380 on blots, analysis, 184, 415 blue copper assays, 188, 285 as electron acceptors for methylamine dehydrogenase, 188, 285 properties, 188, 288 purification from organism 4025, 188, 285 C5 with A49 mutation, preparation, 181, 575 and MI RNA, large-scale reconstitution into RNase P holoenzyme, 181, 581 preparation, 181, 572 purification from cells with rnpAcarrying gene, 181, 573 Ca2+-binding, in platelet surface membrane, identification and characterization, 169, 365 Ca2+-H + antiporter, assay in Neurospora crassa, 174, 669, 674 calmodulin-binding, identification, 184, 451 cAMP-dependent phosphorylation in intact parietal cells, 191,654 capsid, rhinovirus, binding to antipeptide antibodies in immunoblotting, 178, 688 carboxyl terminal residues, amidation, 182, 628 carboxymethylation, 182, 599 carrier conjugation to hapten, 178, 539 vasopressin, 168, 576 coupling of peptides to, 178,701 mediated transport of peptides from brain to blood, assay, 168, 652 cell surface, sialylation, analysis with glycosyltransferases, 179, 94 cellular, flow cytometric assay, 171, 563 centrifugation, 182, 83 252Cfplasma desorption MS analysis, 193, 263

211 channels anion-selective, in human red cells, patch-clamp studies, 173, 120 electrical characterization, 171, 239 function, relationship to membrane structure, 171, 240 messenger-mediated opening, patch clamp studies, 171, 667 number per red cell, 173, 120 single, permeation properties, 171, 683 charge configurations, statistical evaluation, 183, 391 charge runs and periodic patterns, identification, 183, 397 chemical cleavage products, reversedphase HPLC, 193, 389 chemical digestion for MS analysis, 193, 361 chromatofocusing, 182, 380 chromatography on hydroxyapatite columns, 182, 329 cleavage for peptide mapping, 182, 621 in sequencing of N-terminally blocked proteins, 182, 609 cloned, proteolysis in Escherichia coli, minimization, 185, 128 coating of glass micropipet, 191, 344 coding frames, determination, 183,249 coding regions in genomic sequences analytical methods, 183, 169 codon preference and base composition bias, 183, 168 constraints, 183, 164 detection, 183, 163 statistical properties, 183, 164 cofactors and prosthetic groups, 182, 643 colligative properties, 182, 570 in complexes, detection by antibodyinduced mobility shifts, 181, 279 complex family relationships, detection with DNA databases, 183, 124 composition, 182, 568 composition-based analysis, 182, 557 concentration protein stability during, 182, 85 and solute removal, overview. 182.68 conformation comparison to mutants by labeling with :H and 13C, 177, 74, 282

Proteins exchange between states, analysis, 176, 111 heteronuclear NMR analysis, 176, 93 conjugation to glycopeptides, 179, 253 continuous-flow FAB/MS, 193, 222 core from hnRNP particles, purification. 181, 293 snRNP, preparation, 181, 272 crosslinking, as evidence of multienzyme complexes, 182, 547 crystallization, 182, 646 crystals large, growth, 182,657 properties, 182,650 cytoskeletal, removal from erythrocyte membranes, 174, 41 degradation assay with cell-free systems and purified proteins, 186, 499 in intact cells, 186, 493 with purified proteinases, 186, 502 as indicator of oxidative stress, 186. 485 detection in electrophoretic gels, 182. 437 detection of phosphate or sulfate, 193, 482 deuteration for IH NMR, 177, 23 dialysis, 182, 72 direct expression with Escherichia coli trp promoter, 185, 54 disulfide bonds assignment, 168, 96 localization, 193, 374 DNA-binding purification, 182, 521 reverse biochemical analysis, 194, 529 -DNA complexes analysis, 170, 395 crystallization, 182, 658 electrophoretic analysis, 170, 116 footprinting with y-rays, 186, 545 polypeptide domains, mapping and identification, 170, 403 recovery, 170, 392 structure, 31p NMR analysis, 177, 310 -DNA crosslinking in nuclei, 170, 390

Proteins on nucleosomes and soluble chromatin, 170, 389 to partly depurinated DNA, 170, 388 by ultraviolet light, 170, 391 -DNA crosslinks identification in nucleohistone, 186, 540 labeling with [32p]ATP, 170, 406 mapping on DNA fragment, 170, 401 -dodecyl sulfate complexes, precipitation, 170, 395 dynamics interpretation by NMR relaxation measurements, 176, 358 solid-state -'H NMR analysis, 176, 382 electroblotting, 182, 604, 680 electron microscopy, 182, 585 electrophoresis, 182, 82 electrospray ionization MS analysis, 193, 421 elution from gels, 488 engineering, application of hydrophilicity analysis, 178, 584 enzymatic digestion for MS analysis, 193, 361 enzymatic digests, reversed-phase HPLC, 193, 389 epitopes size-exclusion HPLC analysis, 170, 107 tagging and surveillance, 194, 508 eukaryotic, export and secretion from Escherichia coli, 185, 180 exo-, see Exoproteins expressed in heterologous cells, classification, 185, 3 expressed inmammalian cells carbohydrate addition or processing, inhibitors, 185, 589 fatty acylation, 185, 593 gel electrophoresis, 185, 584 glycosylation, analysis, 185, 587 immunoprecipitation, 185, 582 radiolabeling, 185, 579 synthesis, processing, and secretion, 185, 577 tyrosine residues, sulfation, 185, 592 expressed by T7 RNA polymerase-based system

212 analysis, 185, 85 influencing factors, 185, 86 extraneous, removal from aggregated mixed micelles, 191, 601 FAB/MS, 168, 90 footprinting in analysis of antigen-antibody interactions, 178, 751 in epitope mapping, 178, 759 fragmentation nonspecific methods, 193, 383 specific methods, 193, 378 function, effect of N-terminus state, 185, 399 fusion affinity purification, 185, 136 in assay of gene expression products in yeast, 194, 387 localization in Escherichia coli, 185, 132 for minimization of proteolysis in Escherichia coli, 185, 128 from pRIT20 plasmid series, expression and purification, 185, 155 from pRIT30 plasmid series, expression and purification, 185, 157 site-specific cleavages, 185, 140 solubility, 185, 131 G characterization with amiloride, 191, 747 coupled receptors, computer identification, 183, 75 galactose/N-acet ylgalactosamine-binding filter assay, 179, 323 isolation from rat peritoneal macrophages, 179, 324 GAL4, yeast, Western blotting, 185, 304 gel electrophoresis molecular size determination, 182, 581 N-terminal sequence analysis, 182, 604, 613 one-dimensional, 182, 425 for peptide mapping, 182, 615 purity determination, 182, 557 two-dimensional high-resolution, 182, 441 nonequilibrium pH gradient, 182, 446

213 on polyacrylamide minigels, 182, 451 gel filtration molecular weight estimation, 182, 579 overview, 182, 317 in protein concentration and solute removal, 182, 79 purity determination, 182, 562 glucose-H + cotransporter, cDNA clone, identification, 174,401 glutamate residues 3,-carboxylation, 182,634 carboxyl methylation, 182, 632 N~-(3,-glutamic)lysinecrosslink, analysis, 182, 636 -glutathione mixed disulfides, quinoneinduced formation, 186, 630 glycation, nonenzymatic, 182, 645 glycoinositol phospholipid membrane anchors, analysis, 182, 629 glycolipid transfer assay, 179, 559 properties, 179, 564 purification from non-porcine sources, 179, 566 porcine brain, 179, 562 in goblet cell apical membrane, assay, 192, 616 a granule, induced chemotaxis of inflammatory cells and fibroblasts, 169, 233 handling procedures, 182, 84 heat shock encoding yeast genes, induction, 194, 711 high-molecular-weight, charge cluster identification, 183, 399 hsp70, promoter, for cDNA gene expression in mammalian cells, 185,498 hepatitis B virus core, presentation of peptide epitopes on, 178, 669 hepatocyte plasma membrane immunoprecipitation, 191, 833 pulse-chase metabolic radiolabeling, 191, 832 heterologous, s e e Heterologous proteins high-molecular-weight, electrophoretic transfer for immunostaining, 172, 687

Proteins high-performance chromatofocusing, 182, 391 high-performance liquid chromatography on immobilized reactive dyes, 182, 355 for N-terminal sequence analysis, 182, 607 various modes, 182, 392 HIV core, immunoassay, 184, 556 in hnRNP complexes, affinity chromatography, 181, 326 homologies, detection with nucleotide sequence databases, 183, I l I homonuclear Hartmann-Hahn spectroscopy, 176, 159 hybrid denaturing electrophoresis, 194, 487 extraction, 194, 487 a-factor leader-containing, expression in yeast, 185, 412 induction by Trp starvation, 194, 486 production with pATH plasmids, 194. 479 synthesis by polymerase chain reaction, 189, 261 hydrolysis for amino acid analysis, 182, 594 lysosomal, in proximal tubules, 191, 340 hydrophobic chromatography, 182, 339 immobilization on polystyrene balls, 184, 488 immunoblotting, for detection of idiotypes on monoclonal antibodies, 178, 146 immunodetection, 182,683 immunoprecipitation, 182, 688 import by yeast mitochondria, 194, 629 inactivation, causes, 182, 84 inactive, solubilization, 185, 188 -inhibitor contacts, assignment by labeling, 177, 289 internal erythrocyte plasmalemmal, crosslinking, 184, 158 intracellular trafficking, visualization, 184, 379 intron-encoded in yeast mitochondria, genetic studies, 181, 543 ion-exchange chromatography, 182, 309

Proteins iron-sulfur, component of naphthalene dioxygenase system assay, 188, 47 purification, 188, 51 isoelectric focusing overview, 182, 459 in 2D gel electrophoresis, 182, 444 labeling with 15N for ~H and XSNNMR, 177, 44 for peptide mapping, 182, 616 labeling agents, in analysis of anion transporter orientation in membrane, 173, 429 lac repressors in expression of heterologous genes in Bacillus subtilis, 185, 203 headpiece full atomic representation, 177, 241 molecular dynamics simulation, 177, 212 leukocyte surface blotting and avidin-horseradish peroxidase detection, 184, 431 identification, 184, 429 -ligand complexes, intermolecular NOE studies, 177, 355 -ligand interactions, quadrupolar NMR studies, 177, 317 ligands association with band 3 in intact red cell, 173, 518 in solution, 173, 517 purification, 173, 515 reassociation with membranes, 173, 516 solubilization, 173, 515 -lipid interactions, analysis, 172, 632 -lipid interface, mapping, 172, 631 -lipid vesicles, reconstituted, properties, 171, 263 lyophilization, 182, 77 LysR family, detection with D N A databases, 183, 114, 118 malaria circumsporozoite, analysis with program for T-cell epitope recognition, 178, 627 mammalian, expression with insect cells and viruses, 182, 117 mannan-binding,purification from mammalian sera, 179, 310

214 manno-, see Mannoproteins mannose/L-fucose/N-acetylglucosamine-

binding filter assay, 179, 323 isolation from rat peritoneal macrophages, 179, 324 marker, for electrophoretic gel calibration, 182, 439 mass spectrometry by matrix-assisted uv laser desorption/ ionization, 193, 280 molecular weight determination, 193, 354 overview, 193, 351 membrane, see Membrane proteins mercaptan-specific antioxidant assay, 186, 481 properties, 186, 483 purification from yeast, 186, 481 -metal ion binding, quadrupolar NMR study, 177, 325 metallo-, see Metalloproteins metal replication with tungsten, 169, 328 minichromosome-associated, in oivo labeling, 170, 17 modules, detection, 183, 141 molecular weight determination by 252Cfplasma desorption MS, 193, 269 chemical methods, 182, 568 by electrospray ionization MS, 193, 421 by 1-D gel electrophoresis, 182, 439 overview, 193, 441 scattering methods, 182, 584 transport methods, 182, 571 multiple-quantum NMR, 176, 114 multiple sequence alignment and flexible pattern matching, 183, 403 Na+/Ca2÷ exchanger, characterization, 191, 744 Na+/H+ exchanger, characterization, 191, 743 NMR with insensitive nuclei, assignment problems, alleviation, 176, 134 ~SN, measurements, 177, 62 sample preparation for, 176, 78 two-dimensional databases, 176, 62

215 overview, 176, 12 sequential assignment and structural analysis, 176, 50 nonconditional glycosylation defects, m n n mutants with, isolation and characterization, 185,440 N-terminally blocked, sequencing, 182, 609 nuclear cap binding assay, 181, 210 properties, 181, 215 purification from HeLa cells, 181, 212 -nucleic acid affinity, in purification of yeast plasmid chromatin, 170, 35 -nucleic acid crosslinking, reagents, 172, 604 oligomeric structure, characterization by chemical crosslinking, 191, 847 oligomerization analysis with antibodies, 191, 851 correlation with transport, 191, 841 overexpression in E s c h e r i c h i a coli, vectors, hosts, and strategies, 182, 93 eukaryotes, overview, 182, 112 oxidatively modified, carbonyl content assay, 186, 464 oxidative modification, effects on hydrophobicity, 186, 493 molecular structure, 186, 491 proteolytic susceptibility, 186, 491 oxidized, in systems with activated neutrophils or HL60 cells, assay, 186, 575 Pil regulation of glutamine synthetase formation, 182, 807 uridylylation, effect on adenylyltransferase, 182, 799 PADGEM, s e e PADGEM protein pelleting with proteoliposomes, estimate, 173,737 phosphate in, assay, 193, 487 phosphatidylcholine-specific transfer, preparation, 173, 226 phosphorylation analysis, 182, 642 in rabbit platelets, 169, 434 photo-, s e e Photoproteins

Proteins plasma platelet separation from, 169, 33 preadsorption on polymer shunts, 169, 95 precoated polymer surfaces, platelet deposition data, 169, 104 preparation, 169, 93 radiolabeling, 169, 94 platelet, adhesive, dark-field electron microscopy, 169, 326 platelet basic, purification, 169, 225 platelet granule, immunocytochemical localization, 169, 301 -planar lipid membranes, formation, 171, 225 in plant extracts, assay, 182. 182 poly-, s e e Polyproteins polyadenylate-binding characterization, 181, 332 identification, 181, 333 manipulation in S a c c h a r o m y c e s c e r e v i s i a e , 181, 347 purification, 181, 332 Polybuffer removal, 182, 389 postelectrophoretic detection, 182, 490 precipitation with acetone and acid, 182, 592 bulk methods, 182, 292 effects of protein-protein and proteinligand interactions, 182, 304 in enzyme purification, 182, 298 with NaC1 and other univalent-univalent salts, 182, 298 with organic solvents, 182, 296 with polyethylene glycol, 182, 301 in protein concentration and removal, 182, 78 with trichloroacetic acid and deoxycholate, 182, 593 production by secretion from yeast. 194, 491 proteolysis effects on protein purification. 182, 87 for N-terminal sequence analysis. 182. 611 prevention with protease inhibitors. 182, 87 purification laboratory setup, 182, 19

Proteins for N-terminal sequence analysis, 182, 6O2 overview, 182, 9 for peptide mapping, 182, 614 procedures, evaluation, 182, 779 role of detergents, 182, 239 purified encoding gene, cloning, 182, 738 exposure to radicals/oxidants, 186, 489 vectorial crosslinking and biotinylation, 184, 159 purity determination, 182, 555 S-pyridylethylation, 182, 599 -quinone adducts, o-semiquinone radicals from, characterization, 186, 339 radioiodination for peptide mapping, 182, 617 radiolabeling, 182, 721; 191, 345 recombinant C termini and peptides from nonspecific cleavage, identification, 193, 532 disulfide bond analysis, 193, 535 mass spectrometry, 193, 523 production by mammalian cells, optimal cell and culture environment, 185, 567 proteolysis, 193, 521 refolding, 185, 187 reversed-phase HPLC, 193, 522 virus-encoded, detection, 182, 128 related elements, in design of expression plasmid vectors, 185, 29 renal brush border transport, reconstitution and fractionation, 191, 583 retinal-containing, resonance Raman and IR difference spectroscopy, 189, 123 retinaldehyde-binding, cellular form and interstitial retinol-bindingprotein, separation, 189, 327 pigment epithelial, binding of 1l-cisretinaldehyde, 189, 369 retinal photoreceptor, ultrastructural localization, 184, 370 retinoic acid-binding, cellular form assays, 189, 309, 331 gel electrophoresis, 189, 299 purification from human placenta, 189, 330

216 neonatal rat skin and analysis, 189, 307 tissue distribution, 189, 315 types I and II ligand-binding assay, 189, 360 purification from neonatal rat pups, 189, 356 retinoic acid-binding, nuclear form in mammalian cells, identification and analysis, 189, 239 retinoid-binding specificity, 189, 245 retinoid-binding binding properties, 189, 372 cellular assay, 190, 151 in embryonal carcinoma cells, analysis, 190, 148 RNA isolation and Northern analysis, 190, 153 electrophoresis, 189, 302 expressed in Escherichia coli, isotopic labeling for NMR, 189, 514 eye interphotoreceptor matrix assay, 189, 218 partial purification with emphasis on ligand retention, 189, 216 purification, 189, 214 mRNA, in situ hybridization, 189, 286 in parasites isolation, 189, 349 mercurial sensitivity, 189, 351 molecular size profiles, 189, 349 retinol-ivermectin cross-competition for, 189, 352 specificity, 189, 349 radioassay, 190, 142 size-exclusion HPLC, 190, 445 retinol-binding cellular crystallization, 189, 519 gel electrophoresis, 189, 299 and interstitial form, separation, 189, 327 ligand interactions, 189, 514 mammalian, structure-function analyses, 189, 506 radioimmunoassay, 189, 276 tissue distribution, 189, 315 crystal preparation, 189, 283

217

Proteins gene structure and expression, 189, 2O3 interstitial assay, 189, 330 and cellular retinaldehyde- and retinol-binding proteins, separation, 189, 327 cloning, 189, 207 mRNA from retinoblastoma cells, Northern blot analysis, 190, 147 purification, 189, 207 retinoids in, extraction, 189, 328 sequencing, 189,207 tryptic peptides, characterization, 189,210 Western blot analysis, 190, 144 parasite, retinol-ivermectin crosscompetition for, 189, 352 physicochemical properties, 189, 193 purification from plasma, 189, 193 for X-ray crystallography, 189, 282 radioimmunoassay, 189,271 retinol transfer to liposomes and across liposomal membranes, 189, 394 serum, gel electrophoresis, 189, 299 X-ray crystallography, 189, 281 retinoylated, detection, 189, 234 reversed-phase HPLC, 182, 80 in ribonucleoprotein, immunoprecipitared, analysis, 180, 477 ribosomal BLI6 and ELI8, purification, 181, 373 yeast amino acid sequences, 194, 457 core protein preparation, 194, 469 functional analysis, 194, 467 homologies with nonyeast ribosomal proteins, 194, 465 nomenclature correlations, 194, 457 nuclear localization signals, identification, 194, 476 -rRNA interactions, 194, 470 structural analysis, 194, 453 2D minigel electrophoresis, 194, 455 -RNA complexes, isolation and analysis, 1811, 406

and RNA, composition in snRNPs, comparison, 181, 242 -RNA interactions analysis by photoaffinity crosslinking, 1811, 383 genetic analysis, 180, 510 nuclease probing, 180, 454 rotational correlation times, determination by ~3C NMR, 176, 386 secondary structure determination from NMR data, 177, 223 secretion by Bacillus subtilis, overview, 185, 199 parotid acini, assay, 192, 3 l sedimentation equilibrium, 182, 564. 571 sedimentation velocity, 182, 563,574 side chains, ~SN incorporation, 177, 59 single-strand binding, enhancement of electron microscopy of chromatin, 170, 160 size determination, 182, 566 solubility factors, 182, 652 solute-absorbing matrices, 182, 81 solution structure, NMR determination computer programs for, 176, 477: 177. 455 heuristic refinement methods, 177, 218 sorting to yeast vacuole, fidelity, 194, 659 spin-labeled, NMR spectroscopy, 177. 91 spin labeling, 177, 86 stability assessment trials, 182, 86 effect of N-terminus state, 185, 399 enhancement by fusion to/3-galactosidase, 182, 109 by intracellular localization or secretion, 182, 107 with protease-defective hosts. 182, 106 maintenance, 182, 83 solvent conditions for, 182, 85 staining in electrophoretic gels, 182, 477 on nitrocellulose replicas, 184, 433

Proteins preelectrophoretic, with fluorescent stains, 182, 480 quantitation, 182, 487 -steroid conjugates, preparation, 184, 487 storage conditions, 182, 86 structural, as differentiation markers for embryonal carcinoma cells, 189, 345 structure computer analysis, 182, 751 solid-state NMR studies, 176, 242 adjacent peptide planes, relative orientations, 176, 268 peptide backbone structure, 176, 246 in subcellular organelles, assay, 182, 222 subunits, presence of, determination, 182, 566 sulfate in, assay, 193, 487 sulfated, liquid secondary ion MS analysis, 193, 480 superfamilies in organization of Protein Identification Resource, 183, 37 profile analysis, 183, 158 symport Na ÷ with methionine, demonstration, 173, 774 Pi/H +, purification, 173, 734 synthesis inhibition by amiloride and analogs, 191, 748 monoclonal antibody-mutant diphtheria toxic conjugate, 178, 418 Tamm-Horsfall, identification, 187, 368 thiol-containing, biotinylation, 184, 153 thiol status during quinone metabolism, 186, 627 thiyl radical metabolites, 186, 318 three-dimensional structure comparison by rigid-body superposition, 183, 673 derived phylogenetic relationships, 183, 670 multifeature comparison, 183, 679 in solution, NMR analysis, 177, 125 total, in exocytic transport vesicles, analysis, 191, 820 translational motion, measurement by pulse-gradient spin-echo NMR, 176, 418

218 transport identification in ChloreUa, 174, 399 interaction with irreversible inhibitor and substrate, 171, 130 reversible inhibitor and substrate, 171, 117 yeast, analysis by radiolabeling, 194, 669 tryptic fragments, doubly charged, tandem MS, 193, 427 -ubiquitin conjugates degradation, 185, 390 formation, 185, 388 immunological detection in vivo, 185, 392 ultrafiltration, 182, 75 ultraviolet-induced crosslinking to precursor RNA, 181, 68 RNA in vivo, 180, 410 uncrosslinked, depletion from crosslinked samples, 170, 393 unfolded, structural characterization, 176, 458 uniaxially oriented, peptide planes in, solid-state NMR studies, 176, 264 uniporter, mitochondrial Ca2+, transport studies, 174, 75 Ul-specific, UI snRNP particles lacking, chromatography, 181, 254 vacuolar, extracellular, detection by colony immunoblotting, 194, 660 velocity gradient sedimentation, 191, 842 viscosity, 182, 584 visual pigment, phylogeny, 183, 108 vitamin K-dependent, carboxylation, role of DT-diaphorase, 186, 297 yeast activity screening, in gene isolation with Xgtl I system, 194, 234 epitope-tagged immunoprecipitation, 194, 515 radiolabeling in vivo, 194, 513 extract preparation, 194, 819 functional and structural domains, mapping, 194, 533 glycosylation state, analysis, 194, 684 overproduction in genetic studies, 194, 239 tools for, 194, 239

219 secretory, DNA encoding, transcription in vitro, 194, 675 subunit structure determination, 194, 534 synthesis by reverse biochemical methods, 194, 520 in vitro methods, 194, 536 translation, assay, 194, 681 translocation assay, 194, 681 across microsomal membranes in vitro, assay, 194, 675 Protein sequestering assay RNA binding to protein, 181, 277 Proteochondroitin microsomal sulfation, 179,422 properties, 179, 426 Proteoglycans heparan sulfate, in subendothelial extracellular matrix, degradation by platelets, 169, 85 labeling, 179, 430 sulfation, determination, 179, 431 undersulfation by incubation of cells in chlorate-containing medium, 179, 429 sulfate-depleted medium, 179, 428 Proteoliposomes nontransporting, formation, 191, 593 pelletable, protein associating with, estimate, 173, 737 preparation for analysis of membrane protein function modulation by surface potential, 171, 389 conditions for oriented reconstitution, 173, 421 for isolation of phosphate transporter, 173,736 transporting large, formation, 191, 595 Proteolysis anion transporter, 173,423 apoprostaglandin endoperoxide synthase, 187, 479 in cell-free systems, assay, 186, 501 effects on protein purification, 182, 87 in Escherichia coli, minimization by genetic methods, 185, 119 for N-terminal sequence analysis, 182, 611

Protons prevention with protease inhibitors, 182, 87 recombinant proteins, 193, 521 site-specific, fusion proteins from Escherichia coli, 185, 140 susceptibility to, effects of oxidative modification, 186, 491 ubiquitin-dependent, detection and inhibition, 185, 387 during yeast extract preparation, prevention, 182, 168 Protocatechuate 2,3-dioxygenase assay, 188, 95 properties, 188, 99 purification from Bacillus macerans, 188, 97 Protocatechuate 3,4-dioxygenase assay, 188, 83 properties, 188, 87 purification from Brevibacterium fuscum, 188, 85 Protocatechuate 4,5-dioxygenase assay, 188, 89 properties, 188, 92 purification from P s e u d o m o n a s testosteroni, 188, 90 Protonation selective, in protein NMR, 177, 23 Proton current -protonmotive force plots, determination, 174, 90 Protonmotive force -proton current plots, determination, 174, 90 relationship to proton leakage across mitochondrial inner membrane, 174, 85 Protons -alkali metal exchange electroneutrality, assessment, 173, 336 kinetics, 173, 337, 344 net flux studies on A m p h i u m a erythrocytes, 173, 331 pharmacological tests, 173, 336, 343 thermodynamics, 173, 340, 345 -carbohydrate cotransport in Rhodotorula, 174, 629 conductance, determination from state 4 respiration, 174, 89

Protons efflux from human red cells, assay, 173, 181 electrochemical gradient across plasma membrane, determination, 174, 632 exchange with deuterons, in NMR sample preparation, 176, 81 in kinetic analysis of lactate dehydrogenase, 176, 343 extrusion in yeast, analysis, 174, 592 flux rates in parietal cells, assay, 192, 56 - K + symport in Neurospora spherocytes, 174, 654 electrophysiological analysis, 174, 659 ion flux analysis, 174, 662 properties, 174, 664 leakage across mitochondrial inner membrane, assay and relation to protonmotive force, 174, 85 mitochondrial pumps, intrinsic uncoupling, nonequilibrium thermodynamics, 171, 437 Na+-H + exchange in human red cells assay, 173, 181 kinetics, 173, 176 intestinal cell lines, analysis, 192, 381 intestinal membrane vesicles, 192, 399 lymphocytes detection, 173, 777 induced cell swelling, detection, 173, 779 red blood cells, analysis, 173, 292 permeability of nonrespiring mitochondria by swelling, 174, 86 Rhodotorula plasma membrane, 174, 646 -phosphate cotransport system, repression or derepression studies, 174, 608 in protein side-chains, site-specific assignments, 177, 290 pumping in lysosomes and related organelles, assay, 174, 131 spin systems, identification by heteronuclear NMR, 176, 110 translocation by H+oATPase in Neurospora crassa, assay, 174, 668, 674

220 transport ATP-driven, in renal cortical vesicles, 191, 505 driven by Na + gradients, 192, 192, 401 effects of H+,K+-ATPase blockers, 191, 734 in urinary epithelia, regulation, 172, 54 uptake by yeast, and K + outflow, assay in presence of glycine, 174, 625 Protoplasm volume, in determination of nonsolvent space, 174, 244 Protoplasts alteration by plasmolysis, 174, 229 Escherichia coli

fusion, 185, 565 preparation, 185, 564 fusion, in gene transfection for production of chimeric antibodies, 178, 466 guard cell patch-clamp studies, 174, 312 preparation, 174, 313 individual, membrane ion transport measurements, 174, 322 isolation of vacuoles, 174, 555 plant isolation from leaves, 181, 153 suspension cultures, 181, 152 RNA isolation, 181, 155 transfected, pre-mRNA processing, analysis, 181, 148 transfection, 181, 153 transformation, expression vectors for, 181, 154 subceUular fractionation by silicone oil centrifugation, 174, 520 wheat leaf, with short quench times, subcellular fractionation, 174, 526 yeast, preparation, 174, 508 Provitamin A carotenoids supplying, food composition tables for, assessment, 190, 237 Pseudomonas

B13, purification of chlorocatechol 1,2dioxygenase, 188, 125 NCIB 9816, purification of naphthalene dioxygenase, 188, 49 NCIMB 9872, purification of cyclopentanone 1,2-monooxygenase, 188, 78

221

Pyrocarbonic acid diethyl ester

P. acidovorans, purification of gentisate

1,2-dioxygenase, 188, 103 P. aeruginosa

purification of hydroxybenzoate hydroxylase, 188, 139 quinoprotein alcohol dehydrogenase, 188, 34 strain 2x, purification of catechol 2,3dioxygenase, 188, 116 P. cepacia, purification of phthalate dioxygenase and components, 188, 63 phthalate dioxygenase reductase, 188, 65 P. oleovorans, isolation of hydrocarbon monooxygenase, 188, 7 P. putida

purification of muconate cycloisomerase, 188, 128 muconate isomerase, 188, 132 strain FI, purification of toluene dioxygenase, 188, 42 strain ML2, purification of benzene dioxygenase, 188, 56 strain NCIB 12190, purification of cisbenzene dihydrodiol dehydrogenase, 188, 136 P. testosteroni, purification of protocatechuate 4,5-dioxygenase, 188, 90 W6, purification of glucose-6-phosphate dehydrogenase, 188, 347 Psoralen biotinylated, for labeling nucleic acid hybridization probes, 184, 577 crosslinking of chromatin, 170, 155 Psoriasis human, retinoid treatment: murine papilloma test, 190, 382 Pulse-chase analysis heterologous protein expression in Bacillus subtilis, 185, 219 Pulse dampers for pumping systems in HPLC of proteins, 182, 397 Pulse labeling hnRNA in isolated nuclei, 180, 82 pre-rRNAs, 180, 99 prolactin, for assay of secretion from anterior pituitary cells, 168, 259

Pulse radiolysis in analysis of oxygen radicals, 186, 89 Pulse schemes in ~3C off-resonance rotating-frame Ttbased NMR, 176, 394 composite, for decoupling, mixing based on, 176, 156 for reverse correlation in detection of NMR-insensitive nuclei, 176, 136 Pulse sequences for selective excitation in solvent suppression for NMR, 176, 69 Puncture micro-, see Micropuncture Purines -amino acid crosslinks, GC/MS analysis. 193, 850 biotinylated derivatives, probing of adenosine receptors, 184, 668 in DNA, free radical-induced products. GC/MS analysis, 193, 842 free derivatives, GC/MS, 193, 846 modified, in DNA, identification, 193. 853 in RNA, modification by diethyl pyrocarbonate, 180, 374 Pyrene initial ring oxidation by Mycobacterium, enzymatic mechanisms, 188, 151 labeled core nucleosomes, excimer formation in, 170, 205 oxidation, demonstration with fungal lignin peroxidase, 188, 163 S-Pyridylethylation proteins, for amino acid analysis, 182. 599 Pyrimidines -amino acid crosslinks, GC/MS analysis, 193, 850 in DNA, free radical-induced products, GC/MS analysis, 193, 842 free derivatives. GC/MS analysis, 193. 846 modified, in DNA, identification, 193, 853 in RNA, modification by hydrazine, 180, 374 Pyrocarbonic acid diethyl ester in modification interference analysis of

Pyrocarbonic acid diethyl ester reactions with RNA substrates, 180, 369 in structural probing of snRNA, 180, 214, 221 Pyroglutamyl peptide hydrolase interference with glutaminylpeptide cyclase assay, 168, 363 Pyrophosphate -Fe(IV) complex, generation in aqueous solutions, 186, 1l0 Pyrroloquinoline quinone covalently bound, assay, 188, 277 free, assay, 188, 268 isolation, 188, 261 labeling, 188, 266 synthesis, 188, 262 Pyruvate entry into Krebs cycle, competition of enzymatic pathways, 177, 423 Pyruvate carboxylase competition with pyruvate dehydrogenase for pyruvate entry into Krebs cycle, 177, 423 Pyruvate dehydrogenase competition with pyruvate carboxylase for pyruvate entry into Krebs cycle, 177, 423 in intact mitochondria, Ca2÷-sensitive properties, 174, 98 Pyruvate kinase bound reactants and products, equilibrium constant determination, 177, 369 flux rate, 13C NMR assay, 177, 430 Pyruvate synthase catalyzed reactions, role of thiyl radicals, 186, 328

Q Quadrupole reaction cells if-only directional focusing, 193, 162 trajectory stability, 193, 161 translational energy considerations, 193, 166 QUANTA modeling monoclonal idiotypes and anti-idiotypes, 178, 106

222 Quantile plots in analysis of data from in situ hybridization studies, 168, 830 Quenching collisional, fluorescent-labeled nucleosomes, 170, 200 Quin 2 and aequorin, signal comparison, 169, 409 affinity for Ca 2÷, 172, 241 in assay of cytosolic free Ca2÷, 172, 230 platelet cytoplasmic Ca 2+, 169, 399 as calcium-sensitive probe, 168, 271 in control of red cell Ca 2÷ content, 173, 105 entrapment by mammalian mitochondrial matrix, 174, 115 fluorescence calibration for assay of cytosolic free Ca2÷, 172, 240 recording, 172, 238 loading into cytoplasm, 173, 763 intact cells side effects, 172, 253 variables, 172, 233 platelets, cell integrity during and after, 169, 404 localization in cells, 172, 243 Quinacrine in vital staining of yeast vacuoles, 194, 646 Quinohemoprotein alcohol dehydrogenase assay, 188, 36 properties, 188, 38 purification from Comamonas testosteroni, 188, 37 Quinone epoxides HPLC with electrochemical detection, 186, 194 Quinone reductase activity of DT-diaphorase, evaluation, 186, 297 Quinones -amino acid adducts, o-semiquinone radicals from, characterization, 186, 339

223

Radiochemical assay

depletion of cellular and cytoskeletal protein thiols, role of glutathione, 186, 635 glutathione, 186, 630 formation of glutathione disulfides, 186, 630 half-wave reduction potentials, substituent effects, 186, 187 HPLC with electrochemical detection, 186, 180 metabolism, effects on cellular glutathione and protein thiol status, 186, 627 -peptide adducts, o-semiquinone radicals from, characterization, 186, 339 -protein adducts, o-semiquinone radicals from, characterization, 186, 339 Quinoprotein alcohol dehydrogenase assay, 188, 33 properties, 188, 35 purification from Pseuclomonas aeruginosa, 188, 34

R Rabbit anesthesia, 192, 4 Rabies virus monoclonal anti-idiotypic antibody vaccines, 178, 375 Radiation gamma, in footprinting of protein-DNA complexes, 186, 545 inactivation of membrane components, 172, 410 induced damage associated with electron probe X-ray microanalysis, 172, 221 protective effects of ascorbic acid, desferal, and mercaptoethylamine in mice, 186, 693 role of thiyl radicals, 186, 329 ultraviolet, see Ultraviolet radiation Radioactivity in in situ hybridization, assay, 168, 747 specific, tryptic peptides from reductively methylated histones, 170, 383 Radiochemical assay amino acid transport in hepatocyte cultures, 173, 569

avidin, 184, 208 biotin, 184, 208 calcium flux in hepatocytes, 173, 539 /3-carotenoid 15,15'-dioxygenase, 189, 426 cellular retinoic acid-binding proteins, 189, 309 ceramide glycanase, 179, 479 cystine exodus from lysosomes, 174, 160 cytochrome P450 arachidonate oxygenase, 187, 386 cytochrome P450 prostaglandin tohydroxylase, 187, 260 enzymes, substrates, and pathways in isolated nephron segments, 191, 338 erythrocyte anion flux, 173, 58, 63 long-chain alkane oxidation in Acinetobacter, 188, 10 mannosidase IA from rat liver Golgi membranes, 179, 446 membrane-bound NAD+-dependent hexadecanol dehydrogenase, 188, 17 monocarboxylate transport in red blood cells, 173, 302 Na+-K + pumping in muscle rate, 173, 700 stoichiometric ratio, 173, 707 naphthalene dioxygenase components ferredoxin, 188, 49 iron-sulfur protein, 188, 47 nucleotide transport in Golgi complex. 174, 174 phospholipase C-catalyzed phosphoinositide hydrolysis in suspension, 187. 230 platelet-activating factor acetylhyrolase, 187, 345 platelet heparitinase, 169, 343 protein degradation in intact cells, 186, 494 protein kinase C, 168, 348 retinoid isomerase system of eye, 189, 494 retinyl ester hydrolases bile salt-independent, 189, 459 hepatic, 189, 490 RNA cleavage products, 181, 59 RNA ligase and associated activities, 181, 488 RNase BN, 181,431

Radiochemical assay RNase D, 181, 422 RNase T, 181, 427 RNA 3'-terminal phosphate cyclase, 181, 500 serotonin secretion by platelets, 169, 206 streptavidin, 184, 208 sugar transport in red blood cells, 173, 243 tRNA guanylyltransferase, 181, 451 tRNA nucleotidyltransferase, 181, 435 tRNA splicing endonuclease from Saccharornyces cerevisiae, 181, 472 water diffusion across red cell membrane, 173, 193 Radioenzymatic assay catecholamines, 168, 435 Radiofrequency field off-resonance, in t3C NMR determination of protein rotational correlation times, 176, 386 Radioimmunoassay adenosine deaminase-bindingprotein, 178, 174 anti-idiotypic antibodies, 178, 87 anti-PADGEM protein antibodies, 169, 315 cellular retinoic acid-binding protein, 189, 276 cyclic nucleotides in nephron segments, 191, 310 deamido-thyrotropin-releasing hormone, 168, 572 1l-dehydrothromboxane B2, 187, 34 enzymes, substrates, and pathways in isolated nephron segments, 191, 339 estrogen, in granulosa cell aromatase bioassay of FSH, 168, 421 gastrin-releasing peptides, 168, 665 glutaminylpeptide cyclase, 168, 358 histidylproline diketopiperazine, 168, 572 idiotopes carried by antiallergen antibodies, 178, 160 inhibin, 168, 590 interference from passive immunoneutralization, 168, 509 leukotriene C4, 187, 563 leukotrienes, 187, 123 LH-RH precursor, 168, 530, 532 liquid-phase, free ligand binding by antisera, 178, 54

224 neuropeptide Y, 168, 440 peptide processing in vivo, 168, 521 peptidylglycine monooxygenase, 168, 353 phosphomannosyl receptors fi'om bovine testis, 179, 305 physalaemin-related molecules, 168, 448 platelet a2-antiplasmin, 169, 298 platelet PADGEM protein, 169, 318, 322 retinol-bindingproteins cellular form, 189, 276 serum form, 189, 271 solid-phase free ligand, in screening hybridomas, 178, 52 idiotypic and anti-idiotypic antibodies to insulin receptor, 178, 302 solid-state, anti-antimorphine activity, 178, 231 thrombospondin, 169, 262 thyrotropin-releasing hormone, 168, 568 thyrotropin-releasing hormone deamidase, 168, 370 vasopressin metabolites, 168, 389, 394 vegetative tissue lectins, 179, 343 Radioimmunoelectrophoresis platelet von Willebrand factor, 169, 248 Radioimmunoprecipitation monoclonal antibody, in protein footprinting method, 178, 748 Radiolabeling amines and azides with 3H, 191, 765 blotted bands, 184, 426 bombesin, 168, 484 carbohydrate portion of glycoprotein, 194, 690 cell extracts for immunoadsorption, 182, 689 cell surface molecules with glycosyltransferases, 179, 94 cell surface poly-N-acetyllactosamine glycans with [t4C]galactose, 179, 270 crosslinking sites in DNA-protein complexes, 170, 406 DNA at 5' end, 170, 633 DNA primers continuous labeling, 180, 367 at 3' end, 180, 358 at 5' end, 180, 356

225 endothelial cells with [aH]arachidonate, 187, 532 epidermal cells, in assay of retinoid effects on renewal, 190, 77 a-factor, 194, 702 gastrin-releasing peptide, 168, 484 GTP, in screening RNA for group I introns, 181, 537 HeLa cells for analysis of high-molecular-weight kininogen with Na125I, 169,279 histone H5 strong nucleosome binding site, 170, 376 hnRNA and hnRNP, 181, 321 indanyloxyacetic acid IAA-94, 191, 574 ligands for assays of retinoid-binding proteins, 190, 142 lysines binding chromatin DNA, 170, 369 metabolic MDCK filter cultures, 191, 815 pulse-chase, hepatocyte plasma membrane proteins, 191, 832 5-methylcytosine, 193, 859 minichromosomes and associated RNA chains and proteins, 170, 16 mitochondrial translation products for analysis of gene expression, 194, 156 monoclonal antibodies with 3H, 178, 60 mRNA and caps, 180, 174 with 15N, for ~H and 15N NMR, 177, 44 neurohypophyseal hormones, 168, 576 in N-terminal analysis of heterologous proteins secreted by Bacillus subtilis, 185, 221 oligonucleotides, 168, 763 140-base pair nucleosomes, 170, 110 platelets with SICr, 169, 94 effects on ex vioo studies of adhesion to polymeric surfaces, 169, 101 with ~qn, 169, 94, 174 in platelet-rich plasma, 169,457 prelabeling in vivo, 169, 466 with radioactive arachidonate, 187, 590 in suspensions of washed platelets, 169, 458 prolactin. 168, 259

Radioligand binding assay protein 4.1,173, 387 proteins for activity evaluation, 182, 736 for analysis of degradation, 186, 499 endocytosis and lysosomal hydrolysis in proximal tubules, 191, 345 direct iodination, 182, 722 expressed in mammalian cells, 185, 579 indicator molecules, 182, 721 two-stage methods, 182, 732 proteoglycans, 179,430 pulse, see Pulse labeling red cells with p-[3H]azidobenzylphloretin, 191, 778 replicating SV40 chromosomes, 170, 299, 302 RNA for crosslinking to proteins, 180, 413 at 3' end, 180, 157,373 at 5' end, 180, 158, 372 fragments, at 3' ends, 181,586 newly synthesized, with ortho[32p]phosphate, 180, 72 Schizosaccharomyces pombe cells, 194. 820 single-site, circular DNA, 170, 604 snRNAs at 3' end, 181, 24 Tetrahymena intervening sequence at 5' and 3' ends, 181, 562 transfectomas for analysis of chimeric antibody production, 178, 471 whole pancreata, 191, 685 yeast, in analysis of protein transport, 194, 669 yeast glycoproteins, 194, 695 yeast phosphoproteins, 194, 423 yeast proteins in vivo, 194, 513 yeast RNA overview, 194, 423 with 32p in vivo, 194, 408 Radioligand binding assay insulin-like growth factor II receptors, 168, 312 neurotensin receptors, 168, 462 neurotransmitter receptors, 192, 29 thromboxane AJprostaglandin H2 receptors, 187, 397

Radiolysis Radiolysis pulse, see Pulse radiolysis radical generation, principles, 186, 346 Radiometric assay benzoate-CoA ligase, 188, 154 toluene dioxygenase, 188, 40 vasopressin-convertingaminopeptidase, 168, 386 Radionuclides uptake and efflux in whole intestinal cells, analysis, 192, 367 Radioprotection by ascorbic acid, desferal, and mercaptoethylamine, 186, 686 RALIGN program consensus alignment of DNA, 183, 233 Raman spectroscopy in determination of biomembrane surface potential, 171, 367 resonance, retinal proteins, 189, 123 Rapid-reaction methods in assay of ion flux in membrane vesicles, 172, 301 Rat anesthesia, 192, 4 model for cancer chemoprevention by retinoids, 190, 395 vitamin A-deficient, preparation, 190, 229 Rate equations steady-state, for enzyme and carriertransport, generation, 171, 164 RDF2 program evaluation of sequence similarities, 183, 85 Receptors, see also specific receptors blotted, labeling, 184, 424 cell surface amino acid sequencing, 191, 636 binding of radiolabeled ligands, 191, 619 biological activity, 191, 609 covalent crosslinking with ligand, 191, 634 gene sequencing, 191, 637 identification, 191, 609 solubilization, 191, 631 implantation into cells by Sendai virus envelope-mediated fusion, 171, 829

226 mediated Ca 2÷ signaling, analysis, 191, 691 membrane, see interaction with antiidiotypic antibodies, 178, 179 and molecular mimicry, overview, 178, 21 Recombination homologous integrating plasmids, applications, 185, 285 in integrative DNA transformation in yeast, 194, 285 in oivo, in construction of plasmid vectors, 185, 294 intragenic, and haploidization, monitoring in yeast, 194, 96 Reconstitution anion-exchange protein from erythrocyte membrane, 173, 410 ATP-dependent Ca2÷ pump vesicles before purification, 172, 35 bacteriorhodopsin into lipid vesicles, 171, 267 chloride channels, 191, 579 chromatin from purified components, 170, 585 related structures, 170, 598 template allowing transcription, 170, 581 cytochrome P450 arachidonate oxygenase, 187, 392 erythrocytes: preparation of phospholipid membrane-coated cytoskeletons, 173, 392 frozen HL60 cells, 190, 122 glucose transporter from human erythrocytes, 174, 39 Na+-dependent activity, 192, 438 hnRNP complexes, 181, 310 for identification of amino acid transporters, strategies, 173, 159 integral membrane proteins, 182, 516 methylamine dehydrogenase subunits, 188, 245 multienzyme complexes, 182, 550 Na + cotransport after solubilization of transport proteins, 191, 591 natural and hybrid histone octamers, 170, 484

227 nucleosomes cores, 170, 576 core-transcription factor complex, 170, 583 effect of DNA length, 170, 636 at physiological ionic strength, 170, 593 in vitro, in analysis of positioning, 170, 630 phosphate transporter from hepatic mitochondria, 173, 732 renal brush border transport proteins, 191, 583 Sendai virus envelopes, 171, 835 transmembrane, membrane proteins, 171, 265 Rectal gland shark gross anatomy, 192, 756 isolated cells, separation, 192, 765 perfusion, 192, 757 pithing, 192, 762 tubule isolation and perfusion, 191, 183 Rectum electrical measurements, 192, 459 Redox cycling assay free pyrroloquinoline quinone, 188, 277 Reductase NADH oxidoreductase component of methane monooxygenase, assay, 188, 195 Reduction alditol acetates, 193, 560 cholesterol 5,6-epoxides, analysis, 186, 442 cysteine, for amino acid analysis, 182, 598 cytochrome c in assay of HL60 cell differentiation, 190, 126 dye-mediated, 186, 638 free radicals at epidermal surface of skin biopsies, 186, 674 glycolipids by borane, 193, 755 glycoproteins, 193,508 hydroperoxides by iodide, 186, 519 nitro blue tetrazolium, in assay of HL60 cell differentiation, 190, 122

Relaxation times nitroxide in dermal homogenates, 186, 672 -oxidation free radicals, properties, 186, 93 iron: analysis of lipid peroxidation, 186, 457 proteins prior to digestion for MS analysis, 193, 373 Reduction potentials half-wave, quinoid compounds, substituent effects, 186, 187 Reductive cleavage hydroperoxides by cytochrome P450. 186, 278 in linkage analysis of polysaccharides, 193, 573 Reflectance analysis in immunoblot assay of tyrosine monooxygenase, 168, 382 Reflection coefficient determination with pressure probe, 174, 349 from single time course, 173, 220 for direct coupling of water and solute flow in plant cells, 174, 199 in red cell suspensions, measurement. 173,213 transepithelial, in reptilian renal tubules. determination, 191,205 Relaxation matrix analysis complete, in 2D NOE analysis of macromolecules, 176, 169 geometry of aqueous structures, 176. 178 Relaxation rates computer calculation, 177, 458 excess longitudinal in determination of ion binding constants, 171, 312 plots for gramicidin channel interactions, 171, 325 longitudinal, determination by selective saturation recovery, 177, 344 in rotating-frame NOE, 176, 199 Relaxation times based solvent suppression in NMR, 176, 68 in heteronuclear NMR, measurement, 176, 107, 113

Relaxation times in quadrupolar ion NMR, determination, 171, 295 reduction during mixing in HOHAHA, 176, 158 spin-lattice in absence of chemical exchange in quadrupolar nuclei NMR, 177, 319 in determination of protein rotational correlation times, 176, 386 fluorinated ligands, data interpretation, 177, 12 paramagnetic metalloproteins in 1H NMR, 177, 249 phosphorylated amino acids in 3~p NMR, 177, 271 spin-spin in absence of chemical exchange in quadrupolar nuclei NMR, 177, 319 fluorinated ligands, data interpretation, 177, 12 paramagnetic metalloproteins in ~H NMR study, 177, 249 Relay experiments heteronuclear, detection of NMR-insensitive nuclei, 176, 143 Renaturation functionally active membrane proteins after purification, 182, 516 phlorizin binding after solubilization of Na÷-D-glucose cotransporter, 191, 590 polyadenylate-binding proteins, 181, 335 Renin -angiotensin system in juxtaglomerular interstitial cells, assay, 191, 156 Renomedullary cells rabbit, isolation and arachidonate metabolism, 187, 365 Reovirus transcription, in synthesis of capped mRNA in vitro, 180, 168 Reovirus type 3 hemagglutinin polyclonal anti-idiotypic antisera against in characterization of viral receptor on cells, 178, 325

228 development and characterization, 178, 322 structural mimicry by anti-Id antibody, 178, 333 Reovirus type 3 receptors characterization with anti-idiotypic antibody, 178, 325 monoclonal anti-idiotypic antibody development and characterization, 178, 329 structural mimicry of viral hemagglutinin, 178, 333 Repeffusion, see also Peffusion induced injury endothelial cell trigger mechanism, 186, 717 free radical-mediated, in renal transplantation, assay, 186, 748 neutrophil amplification, 186, 714 role of oxygen-derived free radicals, 186, 711 after ischemia heart eicosanoid and PAF release and effects, 187, 616 mitochondrial membrane potential during, measurement, 186, 723 intestine, leukocyte involvement during, 186, 729 related oxidant stress in isolated perfused organs, 186, 752 Replica plating yeast, 194, 18, 185 Replication simian virus 40 chromosomal DNA in vitro, 170, 48 Resealing erythrocyte ghosts, 173, 357 Resins anion exchange preparation for glutaminyl peptidase assay, 168, 362 in isolation of yeast mnn mutants, 185, 449 embedding media for immunoelectron microscopy, selection, 194, 612 epoxy removal from yeast ultrathin sections, 194, 625 section etching methods, 194, 626

229 Resistance electrical, see Electrical resistance multidrug, in heterologous gene selection and coamplification, 185, 555 Resistivity epithelial, large intestine, in vivo measurements, 192, 466 Resonance energy transfer microscopy in visual colocalization of fluorescent lipid probes in liposomes, 171, 850 Resonance-offset effects in rotating-frame NOE, 176, 214 Respiration in gastric parietal cells, measurement, 191, 647 in isolated gastric cells, assessment, 171, 460 state 4, proton conductance from, 174, 89 Respiratory burst stimulation in leukocytes, 186, 552 Restriction endonucleases in analysis of repeated DNA sequences in yeast, 194, 175 cutting of chromatin, 170, 12 digestion of purified DNA, 170, 288 protection, as probe of protein binding, 170, 281 Restriction enzymes database, 183, 26 purification and redigestion of DNA, 170, 330 Rete testis fluid ovine, purification of inhibin, 168, 606 Reticulocytes intact, protein degradation, assay, 186, 494 Retina intermembrane transfer of retinoids, 189, 411 photoreceptor proteins, ultrastructural localization, 184, 370 pigment epithelial cells, short-term incubation, in visual cycle analysis, 190, 3 rat, 4-hydroxynonenal, GC/MS analysis, 186, 399 Retinal binding protein, cellular form

Retinoic acid pigment epithelial, binding of 1 l-cisretinaldehyde, 189, 369 retinol-binding protein separation from, 189, 327 chromatography and characterization. 189, 429 detection: relationship to retinoic acid synthesis, 189, 482 isomers identities, confirmation, 189, 77 preparation and purification, 189, 76 metabolism in epidermal cells: retinoic acid formation, 189, 530 proteins with, resonance Raman and IR difference spectroscopy, 189, 123 tritiated, preparation from tritiated retinol, 189, 532 1l-cis-Retinal

binding to cellular retinaldehyde-binding protein from pigment epithelium, 189, 369 Retinaldehyde, s e e Retinal 1 1 - c i s - R e t i n a l d e h y d e , see 1 l - c i s - R e t i n a l

Retinal oxime isomers, HPLC, 189, 73 preparation from retinal, 189.78 Retinoic acid acylation of protein, 189, 233 administration to amphibians, 190, 190 bead impregnation, 190, 204 binding protein, cellular form assay, 189, 309, 331 gel electrophoresis, 189, 299 purification from human placenta, 189, 330 neonatal rat skin, 189, 307 radioimmunoassay, 189, 276 tissue distribution, 189, 315 types I and II ligand-binding assay, 189, 360 purification from neonatal rat pups, 189, 356 binding protein, nuclear form identification and analysis in mammalian cells, 189, 239 retinoid-binding specificity and characterization, 189, 245 and congeners, local application to chick embryos, 190, 202 derivatization, 189, 107

Retinoic acid effects on epidermal lipid composition, 190, 39 fibroblast attachment to laminin and type IV collagen, 190, 87 limb axes, 190, 191 photodamaged skin, 190, 356 embryonic, derived from maternally administered retinol assay, 190, 317 pharmacodynamics, 190, 320 teratology, 190, 319 formation from retinol and retinal in epidermal cells, 189, 530 HPLC, 189, 50 impregnated beads, implantation in embyronic tissues, 190, 205 induced differentiation of HL60 cells, analysis, 190, 120 labeled, cell incubation with, 189, 239 metabolism by intact cells and cell extracts, 189, 525 regulated murine teratocarcinoma cell genes, expression assays, 190, 131 in skin, effect on angiogenesis, 190, 381 suspension in vegetable oil, 190, 234 synthesis, role of retinal, 189, 482 synthetic analogs handling and characterization, 189, 15 HPLC, 189, 20 all-trans-Retinoic acid and isotretinoin and metabolites, separation, 189, 151 13-cis-Retinoic acid in biological fluids, HPLC, 189, 167 effects on human sebaceous glands, 190, 342 HPLC with automated column switching, 189, 150 inhibition of microsomal lipid peroxidation, 190, 281 metabolites, analysis, 190, 284 pharmacokinetics, 190, 291 therapeutic value, 190, 295 and tretinoin and metabolites, separation, 189, 151 (E)-Retinoic acid deuterated at carbons 4 and 18, synthesis, 189, 17 Retinoic acid receptors anatomy, 189, 228

230 distribution, 189, 228 nuclear assay, 189, 249 binding characteristics, 189, 248 cloning, analysis, and function, 189, 256 -DNA interactions in vitro, 189, 263 domains, analysis by hybrid receptor construction, 189, 261 identification and analysis in mammalian cells, 189, 239 isolation, 189, 248 synthesis in vitro, 189, 263 for testing retinoid activities, 189, 266 Retinoid isomerase eye, assay, 189, 494 retinal pigment epithelium, assay, 189, 503 Retinoid receptors identification as members of steroid and thyroid hormone receptor family, 189, 223 ligands specific for, identification, 189, 225 molecular cloning, 189, 224 Retinoids acidic, HPLC, 190, 168 administration to rats and mice, 190, 234 retinal pigment epithelial cells, 190, 15 antioxidant activity, assays, 190, 273 applied locally to embryos, HPLC, 190, 207 aromatic, in biological fluids, HPLC, 189, 167 binding by adipocyte lipid-binding protein, 189, 366 binding proteins assay with radiolabeled ligands, 190, 142 cellular form assay, 190, 151 binding properties, 189, 372 in embryonal carcinoma cells, analysis, 1911, 148 RNA isolation and Northern analysis, 190, 153 electrophoresis, 189, 302 expressed in Escherichia coli, isotopic labeling for NMR, 189, 516

231 eye interphotoreceptor matrix assay, 189, 218 partial purification with emphasis on ligand retention, 189, 216 purification, 189, 214 mRNA, in situ hybridization, 189, 286 parasite form isolation, 189, 349 mercurial sensitivity, 189, 351 molecular size profiles and specificity, 189, 349 retinol-ivermectin cross-competition for, 189, 352 size exclusion HPLC, 190, 445 binding by retinoic acid-binding proteins, specificity, 189, 245 in blood, HPLC, 189, 50 bound to interstitial retinol-binding protein, extraction and HPLC analysis, 189, 328 in cancer chemoprevention animal models, 190, 395 dose selection, 190, 401 cell lines and cells sensitive to, synopsis, 190, 217 cellular, extraction and analysis, 190, 13 chemoprevention of breast carcinogenesis, 190, 402 conformational analysis, 189, 37 derivatization, 189, 529 deuterated, synthesis, 189, 95 developmental toxicity, evaluation with disposition data, 190,406 dose preparation, 190, 435 down-regulation of squamous cellspecific markers, 190, 42 effects on cell adhesion, 190, 81 cultured Sertoli cells, 190, 71 epidermal renewal, measurement, 190, 76 human sebaceous glands isolated by shearing, 190, 338 photoaged skin, optical profilometry measurement, 190, 360 photodamaged skin, 190, 352, 372 sebaceous glands, 190, 326 sebocyte prolifertion, 190, 334 elemental analysis, 189, 37

Retinoids exchange between lipid vesicles and rod outer segment membranes, 189, 402 gene activation capacity, testing, 189, 266 handling and storage, 189, 19 HPLC, 189, 529; 190, 9 immunomodulatory activity, characterization, 190, 252 induced differentiation of embryonal carcinoma cells, evaluation, 189, 337 induced lipid changes in keratinocytes. 190, 30 inhibition of anchorage-dependent tumor cell growth, 190, 101 anchorage-independent tumor cell growth, 190, 107 interactions with phospholipid membranes ESR studies, 189, 383 optical spectroscopic studies, 189, 373 intermembranous transfer, 189,411 IR spectroscopy, 189, 26 isomers extraction, 189, 60, 72 handling, general precautions, 189, 71 HPLC, 189, 60, 70 radiolabeled, HPLC, 189, 67 lipid vesicles carrying, preparation, 189, 404 melting point determination, 189, 37 metabolism by intact cells and cell extracts, 189, 525 related alcohol dehydrogenase isoenzymes assay, 189, 437 gel electrophoresis, 189, 438 purification, 189, 438 in retinal pigment epithelial cells in vitro, 190, 16 role of various liver cells, 190, 60 metabolites, chromatographic analyses, 189, 533 modulation of phorbol ester effects in skin, 190, 346 molecular weights of selected compounds, 189, 8

Retinoids natural in human skin samples, HPLC, 190, 163 structural characteristics, 189, 3 neutral, HPLC, 190, 165 13C NMR, 189, 31, 33 IH NMR, 189, 26 phagocyte response, 190, 269 pharmacokinetics overview, 190, 291 transplacental and maternal, correlation with teratogenicity during organogenesis, 190, 437 plasma, HPLC, 189, 146 purification, 189, 18 quantitative analysis, 190, 9 radioactive, analysis after isomerase assay, 189, 500 role in epithelial differentiation, 190, 19, 91 keratin synthesis in hamster trachea, 190, 91 rheumatoid arthritis, 190, 175 semiquantitative analysis by direct extraction, 190, 172 sensitive cells and cell lines, synopsis, 190, 217 structural formulas of selected compounds, 189, 8 subacute toxicity, 190, 401 substrates for retinoid isomerase assay, 189, 496 supplemented subjects, immunotrophic methodology, 190, 259 synthesis from retinol and B-carotene, assay and characteristics, 189, 470 synthetic in human skin samples, HPLC, 190, 163 structural characteristics, 189, 3, 43 synthetic manipulation, 189, 15 targeted slow release into chick embryos, 190, 201 teratogenicity mechanistic studies, 190, 418 in vitro testing with micromass limb bud cell culture, 190, 427, in vivo testing, 190, 433 therapeutic application contraindications and risk factors, 190, 301

232 human psoriasis and other keratinization disorders, testing, 190, 382 indications and precautions, 190, 301 murine papillomas systemic treatment, 190, 384 topical treatment, 190, 392 side effects, 190, 302 value, 190, 291 UV spectroscopy, 189, 32, 38 Retinol, s e e Vitamin A Retinol dehydrogenase differentiation from cytosolic alcohol dehydrogenases, 189, 478 NAD+-dependent, detection in hepatic microsomes and characterization, 189, 520 Retinol fatty-acyltransferase assay, 189, 442,446 in bovine retinal pigment epithelial microsomes, assay, 190, 156 Retinol 4-hydroxylase in microsomes and reconstituted systems, characterization, 189, 486 Retinoylation protein, 189, 233 Retinoyl/3-glucuronide in human serum, analysis, 189, 139 HPLC, 189, 140 mass spectrometry, 189, 142 synthesis, 189, 138 Retinyl ester hydrolase bile salt-independent, membraneassociated assay, 189, 460 properties, 189, 461 hepatic, assay, 189, 490 Retinyl esters assay, 189, 89 extraction from serum, 189, 98 and geometric isomers, isocratic adsorption HPLC, 189, 181 HPLC, 189, 81 identification, 189, 90 isomers, HPLC, 189, 74 preparation from retinal, 189, 78 radioactive, analysis, 190, 213 and retinol, separation, 189, 98 reversed-phase HPLC, 189, 85 synthesis by organ-cultured human skin, 190, 210

233 in tissues, assay, 189, 188 total, analysis, 189, 84 Retiny113-glucuronide in human serum, analysis, 189, 139 HPLC, 189, 140 mass spectrometry, 189, 142 synthesis, 189, 137 Retinylidene imminium salts high-field IH and 13C NMR, 189, 112 isomer characterization, 189, 117 preparation, 189, 113 Retroviruses infection of airway epithelial cells, 192, 568 vectors for heterologous gene expression in mammalian cells, 185, 494 Reversed-phase chromatography benzoyl-diacylglyceride molecular species, 187, 210 1 l-dehydro-thromboxane B2, 187, 48 derivatized oligosaccharides, 193,667 high-performance liquid, see Highperformance liquid chromatography, reversed-phase and hydrophobic interaction chromatography, comparison, 182, 340 integral membrane proteins, 182, 513 phospholipid molecular species, 187, 201 RNA, 180, 31 Reverse hemolytic plaque assay hormone release from single cells, 168, 266 identification of anterior pituitary ceils, 168, 148 Reverse transcriptase, see RNA-directed DNA polymerase RHC 80267 inhibition of diacylglycerol conversion to arachidonate, stearate, and glycerol, 191, 690 Rheumatoid arthritis reoxygenation injury in patients, 186, 83 role of retinoids, 190, 175 Rhinovirus binding to antipeptide antibodies in ELISA, 178, 685 capsid protein binding to antibodies in immunoblotting, 178, 688 growth and purification, 178, 683 neutralization, 178,688

Ribonuclease Ill Rhinovirus receptors related synthetic peptide antigens, sitespecific antibodies preparation, 178,677 properties, 178, 683 purification, 178, 681 Rhodanide in determination of yeast membrane potential, 174, 616 Rhodococcus

G-74-2, purification of endoglycoceramidase, 179,490 R. erythropolis, purification of formaldehyde dehydrogenase, 188, 322 R. rhodochrous PNKbl growth and extract preparation. 1518. 26 propane metabolism, cell-free assays. 188, 26 purification of propane-specific alcohol dehydrogenase, 188, 23 Rhodopseudornonas palustris

purification of benzoate-CoA ligase. 188. 156 Rhodotorula glutinis

alkali metal ion transport, 174, 645 carbohydrate accumulation by H + cotransport, 174, 629 carbohydrate transport-catalyzing carriers, number, 174. 638 cell cultivation, 174, 630 plasma membrane effect of N,N'-dicyclohexylcarbodiimide, 174, 647 electrochemical proton gradient, determination, 174, 632 fractions, isolation with cationic silica microbeads, 174, 648 permeability to H + and K ~, 174, 646 substrate accumulation, energetics, 174, 639 Ribonuclease activity in Bacillus subtilis transformants, screening, 185,218 Ribonuclease Ili assay, 181, 422 cloned processing site, analysis, 181, 196 properties, 181, 425 purification from Escherichia coli, 181, 192, 424 test substrates, 181. 190

Ribonuclease A Ribonuclease A, s e e Pancreatic ribonuclease Ribonuclease BN assay, 181, 431 properties, 181, 432 purification, 181, 431 Ribonuclease D, s e e Ribonuclease III Ribonuclease H, s e e Calf thymus ribonuclease H Ribonuclease M5 assay, 181, 371 purification from B a c i l l u s subtilis, 181, 372 Ribonuclease P cleavage activity, assays, 181, 580 holoenzyme reconstitution, 181, 580 purification from E s c h e r i c h i a coli holoenzyme, 181, 571 protein subunits, 181, 572 RNA subunit, 181, 576 RNA moiety, secondary structure, phylogenetic analysis, 180, 227 Ribonucleases active at tRNA 3' terminus, assay, purification, and properties, 181, 421 digestion of pre-RNA, 180, 456 RNA for fingerprinting, 180, 133 pancreatic, s e e Pancreatic ribonuclease partial digestion of terminally labeled RNA, 180, 159 pre-mRNA protection: detection of processing-specific complexes, 181, 86 in probing of RNA interactions with protein and ribonucleoprotein, 180, 454 resistant oligonucleotides, generation from crosslinked RNAs, 180, 431 for RNA sequencing, 180, 156 for RNA structure probing, specificity and properties, 180, 197 Ribonuclease T assay, 181, 427 properties, 181, 429 purification, 181, 427 Ribonuclease TI digestion of internally labeled RNA, 180, 134 in mapping of RNA branches, 181, 185

234 in structural probing of native snRNA, 180, 214, 221 Ribonuclease T2 in determination of RNA branch trinucleotide core composition, 181, 188 resistant ligase product, analysis, 181, 495 Ribonuclease V1 in structural probing of native snRNA, 180, 214, 221 Ribonucleoproteins bound to immobilized pre-mRNA, detection, 181, 109 complexes on nascent RNA, electron microscopy, 180, 481 crosslinked, extraction from UV-irradiated HeLa cells, 180, 414 fibril mapping, 180, 498 heterogeneous nuclear (A2)3B1 tetramer 43S rearrangement product, isolation, 181, 297 insoluble helical filaments, generation, 181, 298 purification, 181, 299 characterization by physical methods, 181, 307 native C-protein tetramer, purification, 181, 300 oligomer binding, 181, 316 particles assembly on defined substrates in vitro, 181, 303 40S, isolation from mammalian cell culture, 181, 294 immunopurification and characterization, 181, 317 proteins in, affinity chromatography, 181, 326 reconstitution, 181, 310 immunoprecipitation with autoantibodies, 180, 468 in mixed sera, antibodies to, relative concentrations, 180, 478 precursors, identification and isolation, 180, 100 -RNA interactions, nuclease probing, 180, 454 RNA-protein interactions in, analysis by photoaffinity crosslinking, 180, 383

235 small from extracts, depletion with autoantibodies, 180, 480 nonmammalian, autoantibody reactions with, 180,479 yeast, preparation and analysis, 194, 405 small nuclear antibodies, effects on polyadenylation in vitro, 181, 67 anti-m3G affinity-purified, anionexchange chromatography, 181, 251 assembly, monitoring, 181, 272 binding to immobilized RNAs, 181, 118 core proteins, preparation, 181, 272 depletion in nuclear extracts by immunoprecipitation, 181, 83 native gel electrophoresis, 180, 442 native, structural probing, 180, 212 particles active in RNA processing, purification, 181, 215 purification with antibodies to modified snRNA nucleosides, 181, 232 RNA in, site-specific degradation, 181, 284 polypeptides applications, 181, 263 isolation, 181,257 protein and RNA composition, comparison, 181, 242 Sm, isolation, 181, 258 U purification and characterization in CsCI gradients, 181,264 structural analysis by in vitro assembly, 181, 273 U1 differential immunoaffinity chromatography, 181, 246 nucleoplasmic, anti-m3G affinity chromatography, 181, 241 particles lacking U l-specific proteins, chromatography, 181, 254 structural and functional criteria, 181, 270

Ribosomes and U5, native, simultaneous isolation, 181, 265 U2 anti-m6A immunoaffinity chromatography, 181, 244 differential immunoaffinity chromatography, 181, 246 nucleoplasmic, anti-m3G affinity chromatography, 181,241 U4/U6 anti-m6A immunoaffinity chromatography, 181,244 nucleoplasmic, anti-m3G affinity chromatography, 181, 241 U5 differential immunoaffinity chromatography, 181, 246 nucleoplasmic, anti-maG affinity chromatography, 181, 241 structural and functional criteria. 181, 270 and UI, native, simultaneous isolation, 181, 265 U7, inactivation by oligonucleotidetargeted RNase H, 181, 85 staining for visualization of yeast nucleolus, 194, 726 structures seen after preparation by Miller method, 180, 501 Ribonucleosides -vanadyl complexes, in preparation of cytoplasm for RNA isolation, 180, 8 Ribosomes binding sites glO-L, in enhancement of foreign gene translation in Escherichia coli, 185, 117 for high-level translation initiation. 185, 89 sequences affecting mRNA translatability, 185, 103 biogenesis, 180, 108 core particles and proteins, preparation. 194, 469 direction to single mRNA species, 185, 103 poly-, see Polyribosomes precursors, isolation and characterization, 180, 96 preparation, 181, 362

Ribosomes rRNA, extraction, 181, 362 yeast, proteins isolation, 194, 454 subunit isolation, 194, 454 Ribozymes Tetrahyrnena

end labeling, 181, 562 in self-splicing and cleavage of RNA, 181, 558 Ricin A chain, conjugate with avidin, preparation, 184, 178 based idiotype-specific immunotoxins for B cell neoplasms preparation, 178, 358 in vitro assay, 178, 361 in vivo applications, 178, 363 in treatment of myasthenia gravis, 178, 451 Rigid-body superposition in comparison of protein structures, 183, 673 Ristocetin in screening of yon Willebrand factor multimer size, 169, 160 R-loops electron microscopy, 180, 252 formation, 180, 250 in RNA mapping, 180, 244 structural interpretation, 180, 254 RNA absorbance melting curves, determination, 180, 304 anchor, biotinylated annealing to pre-mRNA, 181, 106 -pre-mRNA hybrids, assessment, 181, 110 synthesis, 181, 105 binding to protein, protein sequestering assay, 181, 277 biotinylated, affinity chromatography with, 181, 97 branches detection, 181, 183 mapping by primer extension, 181, 184 RNase digestion analysis, 181, 185 trinucleotide core composition, 181, 187

236 capped, long, in vitro synthesis by SP6 and T7 RNA polymerases, 180, 42 capped nucleotides, identification by terminal nucleotide analysis, 180, 161 carrier, preparation, 180, 422 cellular, flow cytometric assay, 171, 561 chemical modification, 180, 373 complementary encoded peptides, in generation of anti-idiotypic antibodies, 178, 63 probes for in situ hybridization histochemical detection of neuropeptide mRNA, 168, 702 for in situ localization of cellular retinol- and retinoic acid-binding protein mRNA, 189, 315 uniformly labeled, in solution hybridization-nuclease protection assays, 168, 645 complex, melting studies, 180, 324 crosslinked ribonuclease-resistant oligonucleotide generation from, 180, 431 2D gel electrophoresis, 180, 426 cytoplasmic, isolation, 180, 6 dideoxy sequencing with reverse transcriptase, 180, 121 digestion by ribonuclease, 180, 133 -DNA crosslinks formation by psoralen method, 170, 157 purification, 170, 157 -DNA heteroduplexes electron microscopy, 180, 252 structural interpretation, 180, 259 -DNA hybridization for primer extension analysis of RNA, 180, 361 principles and reaction conditions, 180, 242 in pulse labeling of hnRNA, 180, 92 for RNA characterization by nuclease S, analysis, 180, 340 in electrophoretic gels, recovery, 180, 23 electrophoretic transfer, 180, 450 end labeling, 180, 193,372 enzymatic hydrolysis, 193, 782 enzymatic sequencing, 180, 154

237 expressed by T7 RNA polymerase-based system, analysis, 185, 85 extraction from nuclei, 180, 12 polyacrylamide gels, 180, 429 fingerprinting interpretation, 180, 150 for RNA processing studies, 180, 130 folding constrained, related data, 180, 279 related computer programs, 180,267 gel electrophoresis, 194, 413 genetic organization, electron microscopic mapping, 180, 239 genomic, preparation for intron mapping, 180, 328 glyoxal removal, 181, 569 group I and I1 introns, sequences and classification, 180, 533 growing chains attached to minichromosomes, in vivo labeling, 170, 16 hairpins, detection, 183, 306 heterogeneous nuclear isolation from nuclei, 180, 91 neuroendocrine peptides, assay, 168, 626 pulse labeling in isolated nuclei, 180, 82 hydrolysates LC/MS analysis, 193, 796 trimethylsilyl derivatives, preparation, 193, 794 identically sized species of different sequence, resolution, 170, 356 immunoprecipitation, 181, 276 intermediate-size, fractionation, 180, 20 internally labeled, RNase digestion, 180, 134 intron mapping, nuclease digestion method, 180, 325 isolation by guanidine hydrochloride method, 190, 134 for neuropeptide gene transcription assay, 168, 684 from protoplasts, 181, 155 labeled fragments, reading, 180, 162 labeling with vaccinia capping enzyme, 181, 179

RNA large, fractionation, 180, 21 lariat, debranching enzyme biochemical requirements, 181, 183 extracts containing, preparation, 181, 183 in RNA structural analysis, 181, 180 mapping by R-loop method, 180, 244 RNA-ssDNA heteroduplex method, 180, 245 as marker of cytosol and rough ER, assay, 182, 221 melting behavior analysis, 180, 322 prediction, 180~ 319 related thermodynamic parameters. 180, 315 messenger assay at cellular and anatomical levels, 168, 808 capped cap characterization, 180, 164 radiolabeling, 180, 174 synthesis in vitro by viral transcription, 180, 166 cellular retinoic acid- and retinolbinding protein, in situ localization, 189, 315 differentially spliced, neurokinin A and substance P, assay, 168, 634 elongation, overview, 185, 43 embryonal carcinoma cell, isolation for analysis of differentiation, 189, 346 erythroid band 3 protein injected X e n o p u s oocytes, anion transport, assay, 173, 453 microinjection into X e n o p u s oocytes, 173, 456 sources, 173, 455 heterologous genes expressed in yeast, analysis, 185, 327 histone, 3' processing in mammalian cell nuclear extracts, 181, 74 immunoglobulin, in analysis of monoclonal idiotypes and anti-idiotypes preparation, 178, 94 sequencing. 178, 98

RNA initiation by Escherichia coli Err7°, 185, 37 interstitial retinol-bindingprotein, Northern blot analysis, 190, 147 keratin, localization, 190, 27 labeling with vaccinia capping enzyme, 181, 179 levels after copper induction of yeast metallothionein promoter, 185, 325 microinjection into MDCK cells, 191, 443 neuroendocrine, nonradioactive visualization, 168, 753 neuroendocrine peptides, assay, 168, 626 neuropeptides, in situ hybridization histochemical detection, 168, 702 newly synthesized, in vivo labeling with ortho[3ep]phosphate, 180, 72 nuclear and cytoplasmic, separation and isolation, 180, 73 oxytocin dot-blot analysis, 168, 407 Northern blot analysis, 168, 405, 407 preparation, 168, 400 in situ hybridization, 168,409 solution hybridization assay, 168, 404 polyadenylylated, selection, 180, 79 polycistronic vectors for cDNA gene expression in mammalian cells, 185, 505 coamplification in mammalian cells, 185, 566 preproenkephalin, detection in rat brain and cultured adrenal medulla, 168, 688 rat intestine, preparation, 191, 441 related factors affecting plasmiddirected expression in Escherichia coli, 185, 25 retinoid-bindingprotein isolation and Northern analysis, 190, 153 in situ hybridization, 189, 286 in single neurons, in situ hybridization studies: data analysis, 168, 822

238 single species, directing of ribosomes to, 185, 103 stability enhancement, 182, 101 measurement, 190, 138 3'-end formation, analysis, 180, 377 3'-end-prncessing-related factors, chromatography, 181, 71 translation enhanced efficiency with two-cistron expression system, 185, 94 in expression of cDNA genes in mammalian cells, 185, 503 high-level initiation, 185, 89 influencing sequences in ribosome binding sites, 185, 103 turnover in cell-free extracts, 181, 202 two-cistron, structural elements, 185, 96 tyrosine monooxygenase, detection in cultured adrenal medulla, 168, 688 vaccinia virus, capping enzyme, purification and use, 181, 170 vasopressin dot-blot analysis, 168, 407 Northern blot analysis, 168, 405, 407 preparation, 168, 400 in situ hybridization, 168, 409 solution hybridization assay, 168, 404 yeast chimeric, decay rates, measurement, 194, 422 decay rates, measurement, 194, 415 preparation by in vitro transcription, 194, 524 translation in vitro, 194, 525, 540 messenger, precursor analysis by transient transfection of HeLa cells, 181, 43 -anchor RNA hybrids, assessment, 181, 110 annealing to anchor RNA, 181, 106 biotinylation, 184, 322 immobilized binding of snRNPs, 181, 118 bound ribonucleoproteins, detection, 181, 109

239 32p-labeled transcripts, preparation, 180, 183 polyadenylation, in expression of cDNA genes in mammalian cells, 185, 502 preparation from mammalian cell nuclei, 180, 69 processing in transfected plant protoplasts, 181, 148 ribonuclease protection: detection of processing-specific complexes, 181, 86 splicing efficiency in yeast, sequence contribution, 181, 122 mammalian extracts active in, preparation and fractionation, 181, 3 nuclear miniextracts efficient in, preparation, 181, 20 SP6, transcription and in vitro splicing, 181, 24 synthesis, 181, 104 transcription termination, in expression of cDNA genes in mammalian cells, 185, 502 micrococcal nuclease pretreatment, 180, 465 modified, cleavage with aniline and gel electrophoresis, 180, 375 modified transcripts, synthesis, 180, 49 nascent transcripts ribonucleoprotein complexes on, electron microscopy, 180,481 splicing, related structures, 180, 504 nuclear flow cytometric assay, 171, 561 isolation, 180, 10 nuclease $1 analysis, 180, 334 oligonucleotide branch points characterization, 180, 185 high-voltage paper electrophoresis, 180, 189 isolation from RNA labeled in vivo with inorganic [32p]phosphate, 180, 178 RNA lariats synthesized in vitro, 180, 182

RNA polyadenylated chromatography, 181, 57 gel analysis, 181, 55 polyadenylation precursor, UV-induced crosslinking to proteins, 181, 68 polyadenylation substrates, preparation, 181, 52 precursor digestion with RNase, 180, 456 immunoprecipitation, 180, 456 protection-immunoprecipitationexperiment, 180,458 radiolabeled, incubation with extract, 180, 456 transcription, 180, 455 preparation with chaotropic agents, 180, 3 for mapping studies, 180, 248 for mass spectrometry, 193, 782 primer extension analysis, 180, 347 probes for high-resolution in situ hybridization histochemistry specifcity, 168,776 transcription in vitro, 168, 772 processing, snRNP particles active in. purification, 181, 215 protected immunoprecipitated fragments analysis, 180, 462 significance, 180, 467 -protecting factor interaction, analysis. 180, 464 protection experiments with antitrimethylguanosine antibodies, 180, 465 and proteins complexes, isolation and analysis, 180. 406 composition in snRNPs, comparison. 181, 242 interactions analysis by photoaffinity crosslinking, 180, 383 nuclease probing, 180, 454 pseudoknots detection, 180, 289; 183, 306 structure and prediction, 180,289 purification by column chromatography, 180, 25. 430

RNA by gel electrophoresis, 180, 14 after transcription and end labeling, 180, 376 from transcription incubations, 170, 356 reactions with, modificatlon interference analysis, 180, 369 in ribonucleoprotein, immunoprecipitated, analysis, 180, 475 -ribonucleoprotein interactions, nuclease probing, 180, 454 ribosomal extraction from ribosomes and polysomes, 181, 362 maturation in Escherichia coli, assay, 181, 355 -ribosomal protein interactions, analysis, 194, 470 5 S, maturation assay in vitro, 181, 365 16 S, maturation assay in vitro, 181, 365 23 S, maturation assay in vitro, 181, 364 26 S, self-splicing intron, secondary structure prediction, 183, 292 ribosomal, precursor purification, 181, 561 self-splicing and enzymatic cleavage by group I intervening sequence, 181, 558 synthesis with phage T7 RNA polymerase transcription system, 181, 560 yeast, isolation and characterization, 180, 96 -RNA interactions, genetic analysis, 180, 510 RNase digests ascending RNA homochromatography, 180, 147 first dimension separation in fingerprinting, 180, 137 kinase labeling, 180, 135 transfer from cellulose acetate to thinlayer plates, 180, 143, 145 RNase H pretreatment, 180, 465 RNase P component MI and C5 protein, large-scale reconstitution into holoenzyme, 181,581 purification, 181, 576

240 5 S, simultaneous sequence alignment and phylogeny reconstruction, 183, 638 satellite, self-cleaving sequences in plant viruses and other sources, compilation, 180, 546 screening for group I introns, 181, 537 secondary analysis, 180, 439 secondary structure computer modeling and display, 183, 318 enzymatic probing, 180, 192 optimal and suboptimal, prediction, 183, 281 phylogenetic analysis, 180, 227 single-stranded, 35S-labeled, as probes for in situ hybridization, 168, 403 size fractionation, 194, 412 small fractionation, 180, 17 self-cleaving, 3-D modeling, 183, 326 purification, 180, 59 synthesis with T7 RNA polymerase, 180, 51 small nuclear anti-Sin immunoprecipitation and 3'end-labeling, 181, 24 3'-end-labeled, chemical modifications and enzymatic cleavage detection via primer extension, 180, 221 direct detection, 180, 214 modified nucleosides, antibodies, in purification of snRNP particles, 181, 232 native, structural probing, 180, 212 sequences, compilation, 180, 521 in snRNP particles, site-specific degradation, 181, 284 splicing in Escherichia coli, genetic and molecular analyses, 181, 521 expression of cDNA genes in mammalian cells, 185, 501 yeast mitochondria, genetic analysis, 181, 539 structure, computer prediction, 180, 262 in subcellular compartments, isolation, 180, 6 substrates, preparation, 181, 275

241 synthesis, inhibition by amiloride, 191, 748 template preparation, 180, 122 terminally labeled partial digestion, 180, 159 preparation, 180, 157 tertiary structure computer modeling and display, 183, 318 enzymatic probing, 180, 192 total extraction from yeast, 1811, 98 purification with guanidinium thiocyahate. 180, 4 T7 RI.I cleavage, 181, 199 mapping and synthesis, 181, 196 transfer cloverleaf structure as computer search pattern, 183, 202 gene transcription, yeast extracts for, 181,439 processing defective substrates, correction, 181, 412 in Escherichia coli, genetics, 181, 395 in yeast, effects of suppressor mutations, 181, 377 yeast extracts for, 181,439 processing-related genes, mutations biochemical consequences, 181, 415 genetic analysis, 181,417 identification, 181,401 suppressor mutants generation, 181, 380 in vitro analysis, 181, 383 in vivo analysis, 181, 388 3' terminus, RNases active at, assay, purification, and properties, 181, 421 tRNAH~L enzymatic addition of guanylate, 181, 451 transfer, precursor encoding gene linked to bacteriophage T7 promoter, construction, 180, 64 end-mature, intron-containing, in vitro synthesis, 180, 63

RNA-directed polymerase synthesis as substrates for RNA processing, 181, 447 by T7 RNA polymerase, 180, 6? yeast cytoplasmic, processing, transacting genes for, analysis, 181. 400 32p-labeled, specific activity in vivo, 180, 116 preparation in vivo, 180, 110 ultraviolet-induced crosslinking mapping, 180, 431 to proteins in vivo, 180, 410 to RNA, analysis: structure-function relationships, 180, 418 viroid and virusoid purification, 181, 585 self-cleavage, characterization, 181, 583 yeast cytoplasmic, isolation, 194, 409 high-molecular-weight, preparation. 194, 398 isolation from ascospores, 194, 401 low-molecular-weight Northern blot analysis, 194, 414 preparation and analysis, 194, 405 mitochondrial, isolation and analysis, 194, 158 nuclear and cytoplasmic, isolation, 194, 409 32P-labeled, preparation in vivo, 194, 4O8 poly(A) *, isolation, 194, 402 prepared, quality evaluation, 194. 404 radiolabeling, 194, 423 total extraction, 194, 406 isolation from vegetative cells, 194, 399 purification, 194, 818 RNA-directed DNA polymerase in dideoxy sequencing of RNA, 180, 121 HIV-I. active, regulated expression in Bacillus subtilis, 185, 210 in structural probing of native snRNA, 180, 221

RNA (guanine-7)-methyltransferase RNA (guanine-7)-methyltransferase activity of vaccinia virus mRNA capping enzyme, assay, 181, 175 RNA guanylyltransferase activity of vaccinia virus mRNA capping enzyme, assay, 181, 172 RNA ligase, see Polyribonucleotide synthase (ATP) RNA polymerase Escherichia coli Ear70 -DNA complex, conformational changes, 185, 40 escape from promoters, 185, 41 promoter recognition and mRNA initiation, 185, 37 SP6, synthesis of long capped transcripts in vitro, 180, 42 T7 delivery to cell by bacteriophage CE6, 185, 65 directed expression of cloned genes in Escherichia coli, 185, 60 induction by IPTG: expression of target DNA, 185, 79 nucleoside triphosphate substrate specificity, 180, 61 in preparation of M1 RNA, 181, 578 synthesis of long capped transcripts in vitro, 180, 42 pre-tRNA, 180, 67 small RNAs, 180, 51 transcription accuracy, 180, 62 type II epitope tagging, 194, 517 fractionation with Polymin P, 194, 517 temperature-sensitive mutants, transcriptional inhibition in yeast, 194, 418 yeast initiation assay, 194, 548 transcription in vitro, 194, 545 type III in transcription from chromatin and cloned gene templates in vitro, analysis, 170, 347 RNASE program graphical display algorithm, 183, 323

242 modeling and display of RNA secondary and tertiary structures, 183, 318 RNA 3'-terminal phosphate cyclase assay, 181, 500 properties, 181, 508 purification from HeLa cells, 181, 504 RNA triphosphatase activity of vaccinia virus mRNA capping enzyme, assay, 181, 175 Rods outer segment membranes, retinoid exchange with lipid vesicles, 189, 402 Roots cells and tissues, ion compartmentation, X-ray microanalysis, 174, 267 pressure, probe method, 174, 218 trans-root potential, measurement, 174, 286 water and solute transport, coupling, 174, 214 Rotary shadowing grids for electron microscopy of SV40 minichromosomes, 170, 173 Rotational correlation time protein, determination by ~3C off-resonance rotating-frame T~ relaxation, 176, 386 Rotofor cells in preparative isoelectric focusing, 182, 475 Rubidium Na~/Rb + stoichiometry in inside-out vesicles, measurement, 173, 378 S6Rb+ influx, effects of furosemide-related diuretics, 191, 790 uptake into isolated cells, assay, 173, 689 uptake by teleost intestine, initial rates, 192, 750 Rubidium iodide and Csl and NaI, positive ion mixture, reference masses, 193, 873 Rubredoxin isolation from Pseudornonas oleovorans, 188, 5 properties, 188, 8

Saccharomyces cerevisiae

243 S Saccharides O-linked from cell surface glycoproteins, characterization, 179, 17 glycopeptides with, CID-MS analysis, 193,709 Saccharomyces aspartic proteinase enzymatic assays, 185, 380; 194, 444 genetic analysis in yeast, 185, 379 overlay test, 194, 442 Saccharomyces cerevisiae a/a and cdc~diploids, sporulation, 194, 101 advantages for genetic and molecular analysis, 194, 3 alleles and mutations at MAT, HMR, and HML, identification, 194, 103 barrier activity, assay, 194, 89 biological pathways, analysis, 194, 245, 249 bud scars, staining with Calcofluor, 194, 732 cell-associated Lucifer Yellow assay, 194, 701 visualization, 194, 700 cell extract preparation for protein surveillance, 194, 514 protein synthesis in vitro, 194, 679 protein translocation studies, 194, 677 studies with regulated GAL4 expression cassette, 185, 304 cell-free translation extract, preparation, 194, 539 cell preparation for thin-section electron microscopy, 194, 602 cell structure, analysis, 194, 248 cell wall chitin, staining with Calcofluor, 194, 732 chromosomal segregation, analysis, 194, 749 colony screens, 194, 497 conjugation with bacteria in transmission of plasmid DNA, 194, 187 constitutive synthesis of heterologous proteins, GAPDH expression cassettes for, 185, 341

DNA complementing, determination of genetic locus, 194, 227 genomic manipulation with plasmid vectors, 185, 280 overview, 194, 6 partially digested, size fractionation, 194, 220 high-molecular-weight, preparation. 194, 218 isolation, 194, 169 repeated sequences, correlation with bands in restriction spectra, 194, 175 shuttle mutagenesis with bacterial transposons, 194, 329 drug resistance, assessment, 185,479 electrocompetent cells, preparation, 194. 183 endocytosis, assays, 194, 697 epitope tagging and protein surveillance. 194, 508 -Escherichia coli shuttle vectors, in generation of yeast mutants from cloned genes, 194, 312 expression of heterologous proteins with ADH2 promoter, 185, 308 hybrid proteins containing c~-factor leader, 185,412 extracts for tRNA gene transcription and processing, preparation, 181,441 s-factor degradation, assay, 194, 709 internalization, assay, 194, 707 leader-directed secretion of heterologous proteins, 185, 408 synthesis, 185,410 genes cloned, in vitro mutagenesis and plasmid shuffling, 194, 302 cloning by complementation, 194, 195 constitutive and inducible expression, vectors for, 194, 389 episomal wild-type, removal by counterselection, 194, 317

Saccharomyces

cerevisiae

mapped list, 194, 835 symbols for, glossary, 194, 830 overexpression, analysis, 194, 239 positional mapping, 194, 57 genetic nomenclature, 194, 8 genetic and physical maps, 194, 827 glycoproteins, analysis, 194, 682 growth, 180, 112; 194, 12 haploidization and recombination, monitoring, 194, 96 heat-shock response, induction and assay, 194, 710 high-copy galactose promoters, regulated GAlA expression cassettes for, 185, 297 hydroxylamine-induced mutagenesis in vitro, 194, 315 isolation of plasmid chromatin, 170, 26 pre-ribonucleoproteins, 180, 100 rRNA precursors and preribosomes, 180, 96 labeling in vivo, 194, 513 literature review, 194, 4 macromolecular fidelity, analysis, 194, 247 macromolecular interactions, analysis, 194, 243 MATa binding of radiolabeled a-factor, 194, 706 isogenic strains containing fusion mutations, 194, 780 MATch, isogenic strains containing fusion mutations, 194, 780 mating and complementation, 194, 18 reaction, assay, 194, 77 meiosis entry into, regulating genes and mutations, identification, 194, 104 monitoring, 194, 94 mutational analysis, 194, 99 membrane structures, visualization, 194, 607 microsomal membranes, protein translocation in vitro, assay, 194, 675 mitochondria DNA, introns in, description, 181, 540

244 genes, analysis and manipulation, 194, 149 RNA splicing, genetic analysis, 181, 539 mitosis, nuclear DNA movement during, digital imaging microscopy, 194, 750 mRNA decay rates, measurement, 194, 415 mutagenesis classical methods, 194, 273 via Ty transposon, 194, 342 mutants allele identification, 194, 317 cell and nuclear fusion-defective assays, 194, 776 with bilateral block, isolation, 194, 782 complementation and dominance tests, 194, 780 genetic screens and selection, 194, 774 isolation, 194, 787, 790 enrichment methods, 194, 276, 279 generation from cloned genes by in vitro mutagenesis and plasmid shuffling, 194, 302 mitochondrial revertants and pseudorevertants, 181, 546 splicing defective, types, 181, 542 mnn, isolation, characterization, and properties, 185, 440 nonmating, mating, 194, 91 nuclear, affecting mitochondrial splicing revertants, 181, 557 screening, 181, 553 Ty-induced, genetic characterization, 194, 352 vacuolar protease-deficient biochemical analysis, 194, 444 genetic analysis, 194, 435 nuclei extract preparation, 194, 546 fractionation, 194, 746 isolation, 194, 735 staining, 194, 99 nucleolar-specific positive stains for optical and electron microscopy, 194, 717

245 organelles enrichment procedures, 194, 666 fractionation, 194,662 marker enzyme assays, 194, 668 partial purification of tRNA guanylyltransferase, 181,453 pheromone production, assay, 194, 84 phosphoproteins, radiolabeling, 194, 423 polyadenylate-binding protein manipulation in, 181, 347 polypeptide transit through secretory pathway, analysis, 194, 662 production of engineered antibodies and antibody fragments in, 178,486 promoters, superimposition of temperature regulation, 185, 351 protein production by secretion, 194, 491 protein synthesis in vitro, 194, 536 protein transport, analysis by radiolabeling, 194, 669 purification of DNA-binding protein OBFI, 182, 524 exocellular superoxide dismutase, 186, 249 mercaptan-specific antioxidant protein, 186, 481 polyadenylate-binding proteins, 181, 341 tRNA splicing endonuclease, 181, 473 random spores, 194, 20 recombinant clones, 194, 241 replica plating, 194, 18, 185 response to pheromones, assay, 194, 86 rho ° strains clone identification, 194, 151 induction, 194, 151 rho- strains induction, 194, 151 synthetic clones, identification, 194, 164 rho ~ strains, mutations affecting single genes, isolation, 194, 152 RNA cellular, extraction, 180, 98 radiolabeling, 194, 423 RNA polymerase II transcription in vitro, 194, 545 run-on transcription, radioassay, 194, 427

Saccharomyces

cerevisiae

secretion of heterologous proteins enhancement by molecular and genetic methods, 185,471 heterologous and homologous signal sequences in, 185, 421 strains for, selection, 185, 472 S- 100 extract coupled transcription and processing assays, 181, 385 preparation, 181, 384 spheroplasts formation and lysis, 194, 663 preparation for isolation of nuclei, 194, 739 isolation of plasmid chromatin, 170, 31, 37 protein translocation studies, 194. 678 RNA extraction, 180, 99 splicing efficiency, pre-mRNA sequence contribution, 181, 122 splicing extract, preparation and fractionation, 181, 89 spore enrichment, 194, 146 sporulation culture media for, 194, 17, 94 monitoring, 194, 94 specific genes interactions, 194, 109 screening and isolation, 194, 108 strains construction, 194, 154 for determination of centromere linkage and meiotic analysis. 194. 43 for gene mapping, 194, 48 overview, 194, 5 for plasmid shuffling, preparation. 194. 307 preservation, 194, 18 protease-deficient, design, 185,385 transcription inhibition with 1,10-phenanthroline, 194, 418 temperature-sensitive RNA polymerase II mutants, 194,419 thiolutin, 194, 418 transformation by electroporation, 194, 182 lithium acetate, 18S, 284; 194, 186 spheroplasting, 194, 186

Saccharomyces cerevisiae translation, sequence and structural requirements, 185, 366 tRNA precursors, preparation in vivo, 18tl, 110 processing, effects of suppressor mutations, 181, 377 Ty mutagenesis, 194, 342 vacuolar proteases activation, 194, 431 biochemical analyses, 185, 379 genetic analyses, 185, 372 influencing factors, 194, 430 interference with yeast studies, prevention, 194, 428 inhibitor cocktails, 194, 433 vegetatively grown, fixation and embedding, 194, 605 Safranine optical probe for membrane potential determination, 172, 78 Salicylate hydroxylation by dye-mediated hydroxyl radical generation, 186, 638 Saline for use in insect Malpighian tubules, 192, 619 Saliva collection from rabbit and rat, 192, 5 composition, 192, 11 Salivary duct short-circuit current, determination in vitro, 192, 16 in vivo, 192, 18 Salivary gland acinar cells, dispersed neuroreceptor-coupled secretory events, analysis, 192, 26 preparation, 192, 26 protein secretion, assay, 192, 31 water transport, related ion fluxes, assay, 192, 33 edema, development during perfusion, 192, 10 perfusion, 192, 10 rabbit, isolation, 192, 8 rat, isolation, 192, 9 secretion stimulation in vivo, 192, 6 in vitro, 192, 11 weight, adjustment for edema, 192, 10

246 Salmonella typhimurium purification of methionine aminopeptidase, 185, 407 Salt absorption by teleost intestine, model, 192, 752 turtle colon, model, 192, 744 extraction of polysomes, 181, 205 flux, effect of p-azidobenzylphlorizin, 191, 776 univalent-univalent, precipitation of proteins, 182, 298 Salt bridges for measurement of ionic permeation properties, 171, 688 Salt gradient dialysis in reconstitution of nucleosome cores, 170, 576 Sandwich assay anti-idiotypic antibodies, 178, 86 Sap sieve tube, collection by aphid stylets, 174, 305 cutting, 174, 302 Saponification leukotriene A4 methyl ester, 187, 288, 557 methyl epoxyeicosatrienoic acids, 187, 363 Saponin permeabilization of pancreatic acinar cells, 192, 283,292 Sarcoplasmic reticulum rabbit muscle, Ca2*-ATPase crosslinking, 172, 606 SCH 28080 effect on gastric gland preparations, 191, 729 inhibition of gastric acid secretion, effect of neutralization, 191, 731 Schistosomiasis immunoprophylaxis, anti-idiotypic antibodies in, 178, 390 Schizosaccharomyces pombe antibody staining, 194, 823 biology, overview, 194, 796 cell growth, 194, 799 cell wall digestion, 194, 822 chromosomal DNA preparation, 194, 817 culture media, 194, 800 fixation, 194, 822

247 genes cloning by gap repair, 194, 815 disruption and replacement, 194, 816 genetic analysis classical methods, 194, 803 molecular methods, 194, 795 immunottuorescence studies, 194, 822 mutagenesis, 194, 805 nuclei isolation, 194, 743 staining, 194, 821 plasmids, 194, 806, 810 protein extracts, preparation, 194, 819 radiolabeling, 194, 820 spores, random, analysis, 194, 804 strains diploid, isolation, 194, 804 reisolation and phenotype testing, 194, 798 total RNA preparation, 194, 818 transformation, 194, 808 Scintigraphy in analysis of Hqn-labeled platelet turnover and organ distribution in v i v o , 169, 174 Scintillation counting liquid, s e e Liquid scintillation counting Scintillation detectors in mass spectrometry, 193, 68 Scrape loading permeabilization of mammalian cells to macromolecules, 171, 868 SDS, s e e Sodium dodecyl sulfate Sea urchin embryo, nuclei active, preparation, 181, 34 functionally intact, isolation, 170, 435 isolation with detergent, 170, 437 purification of histone subunits, 170, 461 sperm nuclei, isolation, 170, 438 purification of histone subunits, 170, 460 Sebaceous glands effects of retinoids, 190, 326 human, isolated by shearing, effects of isotretinoin, 190, 342 retinoids, 190, 338 size, quantitation, 190, 330

Sedimentation Sebocytes growth suppression assay, 190, 335 growth in vitro, 190, 336 isolation and culture conditions, 190, 335 proliferation, effects of retinoids, 190, 334 Sebum secretion, inhibition by retinoids, 190. 327 Secretin immunoassay with charcoal, 182, 711 Secretory granules characterization, 174, 169 physiologically responsive, isolation from exocrine tissues, 174, 162 Secretory leukocyte protease inhibitor refolding, 185, 190 Sedimentation differential enterocytes, 192, 344 microsomes and mitochondria, 172.23 subcellular membranes, 172, 19 immuno-, s e e Immunosedimentation isopycnic anterior pituitary cells, 168, 64 in purification of lactotropes and somatotropes from adult male rats, 168, 66 somatotropes from 14-day-old female rats, 168, 68 polyadenylation-specific complexes, 181, 63 reconstituted hnRNP complexes, 181. 311 in sucrose gradient analysis of nucleosomes, theory, 170, 56 unit gravity, in cell separation, 171, 492 velocity gradient density gradient purpose in, 171. 468 proteins, 191, 842 two-step, in lactotrope purification from adult female rats, 168, 69 velocity, at unit gravity anterior pituitary cells, 168, 63 in cell separation, 171, 497 mammalian cells, 171, 501 in purification of lactotropes and somatotropes from adult male rats, 168, 66 somatotropes from 14-day-old female rats. 168, 68

Sedimentation coefficients Sedimentation coefficients erythrocyte band 3 anion transporterTriton X-100 complex, 172, 700 membrane protein-detergent complexes, 172, 699 prostaglandin E~-Triton X-100 complex, 172, 703 in sucrose gradients, determination, 170, 64 Sedimentation equilibrium in determination of protein purity, 182, 564 protein size and presence of subunits, 182, 571 Sedimentation velocity in determination of protein purity, 182, 563 protein size, molecular weight, and presence of subunits, 182, 574 Seeds legume, coats, assay for phloem unloading, 174, 310 Selected-ion monitoring applications, 193, 89 in detection of ion currents, 193, 86 dwell times, selection, 193, 98 with GC/MS in assay of free radical-induced products in DNA and nucleoprotein, 193, 855 oxidative DNA damage, 186, 530 identification of DNA-protein crosslinks in nucleoprotein, 193, 855 modified bases in DNA, 193, 853 m/z values, selection, 193, 95 operation mechanics, 193, 99 partially methylated alditol acetates, 193, 572 quantitative computation from, 193, 104 Selective saturation recovery in determination of longitudinal relaxation rate, 177, 344 Seminal vesicles ovine, microsomes preparation, 187, 247 purification of prostaglandin H2, 187, 248

248 o-Semiquinone radicals from catechol estrogens, spin stabilization, 186, 341 complexed and uncomplexed, spectral parameters, 186, 337 generation and detection, 186, 337 production in biological systems, ESR spectroscopy, 186, 333 from quinone adducts with amino acid, peptide, and protein, characterization, 186, 339 secondary, identification, 186, 338 spin stabilization, 186, 334, 341 Sendal virus envelope mediated fusion for carrier and receptor implantation into cells, 171, 829 reconstituted fusogenic activity, estimation, 171, 839 fusogenic, preparation, detergent selection for, 171, 842 reconstitution, 171, 835 permeabilization of mammalian cells to macromolecules, 171, 865 virions, intact fusogenic activity, estimation, 171, 832 propagation and purification, 171, 831 Sepharose -avidin, in identification of complement C3 acceptors, 184, 624 avidin immobilization, 184, 185 matrices, immobilization of IgGs, 181, 238 Sepharose CL-4B activation by 4-nitrophenyl chloroformate, 184, 81 Serine spin system identification, 177, 138 Serine carboxypeptidase yeast and biocytinamide, C-terminal biotinylation of proteins, 184, 160 enzymatic assay, 185, 384; 194, 448 filter assay, 194, 659 genetic analysis, 185, 374 overlay test, 194, 437 well test, 194, 439

249 Serine-glyoxylate aminotransferase assay in crude extracts of methylotrophs. 188, 362 Serine hydroxymethyltransferase, s e e Glycine hydroxymethyltransferase Serological assays anti-idiotypic antibodies, 178, 75 Serotonin 14C-labeled, release from rabbit blood, assay, 169, 470 hemorrhage-induced release in supraoptic nucleus, microdialysis assay, 168, 201 3H-labeled, release, in bioassay of PAF, 187, 130 secretion by platelets assays, 169, 196, 435 effects of diacylglycerol and Ca z+ ionophore, 169, 436 fluorimetric assay, 169,208 radioactive assay, 169, 206 Sertoli cells rat, cultures, preparation and effects of retinoids, 190, 71 Serum, s e e a l s o Blood, Plasma bovine effect on quirt2 loading into intact cells, 172, 238 purification of conglutinin, 179, 319 exocellular superoxide dismutase, 186, 249 mannan-binding protein, 179, 319 carcinoembryonic antigen, assay, 184, 507 a-carotene and other nutrients, simultaneous HPLC, 189, 155 /3-carotene liquid chromatographic assay, 189, 172 and other nutrients, simultaneous HPLC, 189, 155 extract, glucuronides, HPLC, 189, 140 human oligosaccharide analysis, 179, 55 purification of mannan-binding protein, 179, 312 oligosaccharides, 179, 55 platelet-derived growth factor, 169, 213

Shear flow retinyl and retinoyl/3-glucuronides, analysis, 159, 139 immunoassay samples, measurements. 182, 705 lycopene and other nutrients, simultaneous HPLC, 189, 155 mixed, antibodies to ribonucleoproteins, relative concentrations, 180, 478 purification of anti-m3G and anti-m6A antibodies, 181, 237 rabbit polyclonal antibody preparation, 182, 665 purification of mannan-binding protein, 179, 321 rat, purification of mannan-binding protein, 179, 317 retinol liquid chromatographic assay, 189, 172 and other nutrients, simultaneous HPLC, 189, 155 and retinyl esters, extraction, 189, 98 a-tocopherol, liquid chromatographic assay, 189, 172 tocopherols and other nutrients, simultaneous HPLC, 189, 155 vertebrate, detection of JE antibody. 184, 497 Serum albumin bovine biotinylation, 184, 227 coupling of glycopeptide, 179, 256 effect on quin2 loading into intact cells, 172, 238 rotational correlation times, determination by 13C NMR, 176, 414 human -hygromycin phosphotransferase fusion cassette, construction, 185, 476 -invertase fusion cassette, construction, 185,476 secretion by yeast, enhancement by molecular and genetic methods, 185, 471

Shear flow effect on thromboembolization, 169, 99 rate control during perfusion, 169, 58 rate effects on platelet-surface interactions, 169, 41

Shearing Shearing related forces, in cell disruption, 172, 5 sebaceous glands isolated by, effects of retinoids, 190, 338 skin, in isolation of sebaceous glands, 190, 339 Sialate 9-O-acetylesterase assay, 179, 411 properties, 179, 413 purification from rat liver, 179, 412 Sialylation cell surface proteins, analysis with glycosyltransferases, 179, 94 a-2,6-Sialyllactosamine synthesis, 179, 275 a-2,6-Sialyllactose synthesis, 179, 275 Sieve tube sap, collection by aphid stylets, 174, 305 cutting, 174, 302 Sieving parietal cells, 187, 510 Silanization micropipettes, 172, 142 Silica, see Silicon dioxide Silicon dioxide cationic microbeads, in isolation of Rhodotorula plasma membrane fractions, 174, 648 Silicone oil microcentrifugation in, in assay of Mg2÷ transport in suspension culture, 173, 551 Siliconization glass micropipet, 191, 344 Silver staining proteins for assay, 182, 487 in electrophoretic gels, 182, 438, 482 stain properties, 182, 484 yeast nucleolar-specific for electron microscopy, 194, 725 for light microscopy, 194, 721 Simian virus 40 chromatin, scanning transmission electron microscopy, 170, 175 chromosomes, replicating affinity isolation, 170, 41 enzymatic digestion, 170, 297

250 extraction, 170, 300 incorporation of Bio-19-SS-dUTP, 170, 45 preparation and radiolabeling, 170, 299 infection of CV1 cells, 170, 15, 46 minichromosomes electron microscopy, 170, 166 preparation and characterization, 170, 14 nucleosomes, electron microscopy, 170, 176 replication forks, chromatin structure at, 170, 291 transformed'/3 cells, establishment and culture, 192, 227 vectors for heterologous gene expression in mammalian cells, 1115,488 Similarity functions in identification of homology domains, 183, 135 Similarity scoring aggregates, for sequence comparison, 183, 141 with mutation data matrix applications, 183, 333 limitations, 183, 341 Size-exclusion chromatography high-performance, see High-performance liquid chromatography RNA, 180, 40 Size fractionation partially digested yeast genomic DNA, 194, 220 RNA on DEAE resins, 194, 412 sucrose density gradient, in preparation of clone libraries in YAC vectors, 194, 261 Skeletal muscle reperfusion injury, role of oxygenderived free radicals, 186, 717 Skeleton malformations, examination and evaluation, 190, 420 rat fetus, staining with alizarin red S, 190, 420 Skin, see also specific tissues frog, and toad bladder, comparison for Na + transport studies, 192, 689

251 human natural and synthetic retinoids, HPLC, 190, 163 organ cultures preparation, 190, 211 synthesis of 3,4-didehydroretinol and retinyl and 3,4-didehydroretinyl esters, 190, 210 photoaged, effects of topical retinoids, optical profilometry measurement, 190, 360 psoriasis and other keratinization disorders, retinoid therapy, testing, 190, 382 shearing, in isolation of sebaceous glands, 190, 339 surface impressions image analysis results, correlation with clinical gradation, 190, 366 preparation, 190, 362 murine biopsy, free radical reduction at epidermal surface, 186, 674 histology, 190, 355 homogenates, nitroxide reduction, analysis, 186, 672 papilloma, in testing of retinoid therapy of keratinization disorders, 190, 382 photodamaged, effects of retinoids, 190, 352,372 preparation from hairless animals, 186, 671 wrinkle measurements, 190, 356 phorbol ester effects, modulation by retinoids, 190, 346 preservation for transplantation, effect of ischemia-reoxygenation injury, 186, 81 rat, neonatal homogenate preparation, 189, 308 purification of cellular retinoic acidbinding protein, 189, 307 reperfusion injury, role of oxygenderived free radicals, 186, 717 Smith assay for proteins, overview, 182, 60 Smooth muscle porcine aortic, membrane preparation, 187, 400

Sodium rat vascular, culture preparation, 187, 400 Snail juice digestion of ascus wall, 194, 32 Sobemoviruses probable, satellite RNAs, self-cleaving sequences, compilation, 180, 547 Sodium active transport in turtle colon, 192, 736 amiloride-sensitive reabsorption by reptilian distal tubules, 191,220 assay with sodium-binding benzofuran isophthalate, 192, 73 Ca2+-2Na + exchange, analysis, 174, 81 coupled solute transport, characterization, 191, 744 efflux from human red cells, assay, 173, 188 fluxes assay with sodium-binding benzofuran isophthalate, 192, 78 through sodium pump, assay, 173, 92 gradients with defined magnitude, creation in intestinal epithelial cells, 192, 336 driven proton transport, analysis, 192, 401 intracellular, assay in intestinal epithelial cells, 192, 337 Li+/Na + exchange in human red cells. assay, 173, 184 22Na+ efflux from isolated cells, assay, 173, 694 influx, effects of furosemide-related diuretics, 191, 790 Na+/Ca ÷ exchange in cardiac sarcolemmal vesicles, charge movements during, 172, 155 red blood cells, analysis, 173, 292 Na+/H + exchange in human red cells assay, 173, 181 kinetics, 173, 176 induced cell swelling, detection, 173, 779 in intestinal cell lines, analysis, 192, 381

Sodium in intestinal membrane vesicles detection, 192, 399 regulation, 192, 404 in lymphocytes, detection, 173, 777 in red blood cells, analysis, 173, 292 in turtle colon basolateral membrane, 192, 743 Na+/K+/CI- cotransport in intestinal cell lines, pathway, 192, 371 system identification and purification, 191, 786 Na+/K+(Rb +) stoichiometry in inside-out vesicles, measurement, 173, 378 Na+/Li ÷ exchange in human red cells, assay, 173, 184 Na+/Na + exchange in human red cells, assay, 173, 184 intestinal membrane vesicles, analysis, 192, 402 Na+/P~ coupling ratio in inside-out vesicles, measurement, 173, 378 net extrusion from muscle, 173, 697 pH gradient-driven uptake by intestinal membrane vesicles, 192, 399 platelet plasma membrane gradient, effect on Ca transport, 169, 383 radioactive and nonradioactive, loading of red cells, 173, 187 role in amino acid transport in red cells, 173, 143 secretion by Malpighian tubules, analysis, 191, 176 in sweat gland tissue, assay, 192, 596 transepithelial transport, toad urinary bladder as model, 192, 683 transport in intestinal cell lines, pathways, 192, 381 kidney, effects of furosemide-related diuretics, 191, 785 uptake by amiloride-blocked bladder membrane vesicles assay, 172, 159 kinetics, 172, 161 isolated renal cells, system identification, 191, 399 teleost intestine, initial rates, 192, 750

252 Sodium arachidonate 3H-labeled, preparation, 187, 590 Sodium-binding benzofuran isophthalate in assay of Na + levels, 192, 73 Na ÷ fluxes, 192, 78 fluorescence spectra, 192, 74 Sodium bisulfite modification of DNA, 184, 602 Sodium channels amiloride-blockable, ion fluxes, assay, 172, 158 epithelial, characterization, 191, 742 functional sites, mapping with synthetic peptide antibodies methods, 178, 720 overview, 178, 714 in turtle colon basolateral membrane, analysis, 192, 743 voltage-gated, characterization, 191, 745 Sodium chloride precipitation of proteins, 182, 298 secretion in glomerular nephrons, 191, 195 -sodium citrate, oligonucleotide hybridization in, 182, 750 transport pathways in intestinal membrane vesicles, 192, 389 Sodium citrate -sodium chloride, hybridization with oligonucleotide probes, 182, 750 Sodium dodecyl sulfate induced membrane protein dissociation, resistance, 191, 850 in molecular weight determination of membrane protein constituent polypeptides, 172, 520 -polyacrylamide gels calibration with marker proteins for electrophoresis, 182, 439 casting, 182, 431 casting and running for mini-2DPAGE, 182, 453 one-dimensional, casting and running for 2-D electrophoresis, 182, 446 -protein complexes, KC1 precipitation, 170, 395 second-dimensional gels preparation for electrophoresis of core histones, 170, 547

253 DNA, 170, 130 histone HI, 170, 539 staining, 170, 132 solubilization of Staphylococcus aureusbound antigen-antibody complexes, 182, 697 Sodium hydroxide -borax buffer stock solutions, 182, 37 -glycine buffer stock solutions, 182, 37 in permethylation of oligosaccharides, 193, 650 Sodium iodide and Csl and Rbl, positive ion mixture, reference masses, 193, 873 Sodium pump, see ATPase, Na+,K +Solid-phase assay sequential, t251-1abeled avidin-based, for biotin, 184, 224 Solubility amiloride analogs, 191,749 detergents, pH dependence, 182, 256 proteins, influencing factors, 182, 652 Solubilization anion-exchange protein from erythrocyte membrane, 173,416 antigen-antibody complexes bound to Staphylococcus aureus, 182, 697 arachidonate-CoA ligase from cell membranes, 187, 238 band 3 protein, 173,499, 501 bilitranslocase, 174, 55 brush border membrane proteins, 191, 587 cell surface receptors, 191,631 chloride channels, 191, 575 chromatin, 170, 143, 146 gonadotropin-releasing hormone receptors, 184, 288 inactive recombinant proteins, 185, 188 integral membrane proteins, 182, 505 lipids organic solvents for, selection, 171, 210 for preparation of unilamellar vesicles, 171, 205 liposomes for preparation of unilamellar vesicles, 171, 205 membrane proteins with nonionic surfactant, 172, 519 microsomal membranes, 182, 231

Solutes Na+-D-glucose cotransporter, phlorizin binding after, renaturation, 191, 590 native membrane proteins, 182, 253 in preparative-scale transport-specific fractionation, 172, 46 protein aggregates in Escherichia coli. 182, 264 protein ligands, 173, 515 pulmonary microsomes, 187, 254 selective, band 3 protein, 173, 501 stripped ghosts, 173, 500 transport proteins, Na ~ cotransport reconstitution after, 191, 591 for 2D-PAGE of proteins, 182, 443 Solute potential cell sap, measurement, 174, 242 Solutes, see also specific solutes active transport in insect Malpighian tubules, 192, 624 bidirectional transepithelial fluxes in renal cell lines, assay, 191, 436 crossing of Malpighian tubule walls, route determination, 192, 626 efflux from erythrocyte ghosts, kinetics. 173, 360 fluxes in monolayer cell cultures, measurement devices, 171, 133 intracellular levels in isolated per[used renal tubules, 191, 366 movement, microcomputer modeling, 172, 280 Na+-coupled transport, characterization. 191,744 passive permeability of plant cells, measurement by endoosmotic permeation, 174, 252. 262 partial concentration method, 174. 256 relations dynamics, 174, 187 limitation of plant growth, 174. 213 transport in defined renal cells in vitro, 191. 418 in insect Malpighian tubules, analysis and control, 192, 617 transmembrane, in renal cell lines. assay, 191,430 and water, flow in plant cells, direct coupling, 174, 199

Solution-phase assay Solution-phase assay epitope specificities and affinities, 184, 518 Solutions for analysis of erythrocyte anion fluxes, 173, 54 aqueous, Fe(IV) and Fe(V) complexes, generation, 186, 108 detergent, associated critical phenomena, 172, 530 enzymes for DNA and RNA hydrolysis, preparation, 193, 785 HCO3/CO2, pH control, 192, 42 histones, oligomeric state, determination, 170, 557 metal-free, preparation, 186, 121 micellar, preparation, 189, 419 organic, photochemical reactions in, 172, 646 for preservation of intracellular function after breach of plasma membrane, 171, 818 proteins, structure distance geometric analysis, 177, 193 NMR determination computer programs for, 176, 477; 177, 455 by heuristic refinement, 177, 218 three-dimensional analysis, 177, 125 WEB 2086 and 2170, preparation and stability, 187, 456 Solvents, see also specific solvents for affinity chromatography of proteins, selection, 182, 360 carrier, composition for continuous-flow FAB/MS, 193, 217 for gel filtration of proteins, 182, 322 for HPLC of proteins, 182, 396 organic for lipid solubilization, selection, 171, 210 precipitation of proteins, 182, 296 suppression for NMR, 176, 64 volatile, lipid solution injection for unilamellar vesicle preparation, 171, 207 Somatic cells microinjection of antibodies, 170, 245

254 Somatostatin release from hypothalamus effects of neurotransmitters and neuropeptides, 168, 728 in vitro analysis, 168, 719 in vivo analysis, 168, 716 Sonication in production of small unilamellar vesicles, 171, 202 ultra-, see Ultrasonication Spacer arms in affinity chromatography of proteins, 182, 360 Spectral density in ~3C off-resonance rotating-frame Ttbased NMR, functions, 176, 391 relationship to relaxation parameters, modeling, 176, 363 Spectrin purification from erythrocyte membranes, 173, 384 Spectrophotometry, see also specific techniques

acetol dehydrogenase, 188, 31 acetyl-CoA-independent phosphoenolpyruvate carboxylase, 188, 363 alcohol dehydrogenase cell-free methods, 188, 29 isoenzymes in retinoid metabolism, 189, 437 propane-specific form, 188, 21 soluble NAD(P)+-dependent forms, 188, 15 alcohol oxidase, 188, 421 aldehyde dehydrogenase cell-free methods, 188, 29 from Hyphomicrobium X, 188, 328 soluble and membrane-bound NAD(P)+-dependent forms, 188, 19 aldehyde dehydrogenase (acceptor), 188, 323 amine oxidases, 188, 427 arachidonate 5-1ipoxygenase from porcine leukocytes 187, 339 potato, 187, 297 cis-benzene dihydrodiol dehydrogenase, 188, 134 benzene dioxygenase, 188, 54

255 blue copper proteins, 188, 285 /3-carotene incorporated in micelles, 189, 420 catalase, 188, 463 catechol 1,2-dioxygenase, 188, 123 catechol 2,3-dioxygenase, 188, 116 chlorocatechol 1,2-dioxygenase, 188, 123 cyclohexanone 1,2-monooxygenase, 188, 72 cytochromes c, 188, 290 cytochromes cH and CL soluble forms, 188, 299 stopped flow methods, 188, 304 formaldehyde dehydrogenase with cofactor requirement, 188, 321 from methylotrophic bacteria, 188, 318 from methylotrophic yeast, 188, 455 formaldehyde dehydrogenase (glutathione), 188, 315 formaldehyde transketolase, 188, 435 formate dehydrogenase, 188, 459 glucose-6-phosphate dehydrogenase from Arthrobacter globiformis, 188, 339 Methylobacillus flagillatum, 188, 335 Pseudomonas W6, 188, 346 glycerate kinase, 188, 362 glycerone kinase, 188, 451 3-hexulose-6-phosphate synthase, 188, 392 hydroxybenzoate hydroxylase, 188, 139 hydroxypyruvate reductase, 188, 373 lignin peroxidase, 188, 160 malyl-CoA lyase in crude extracts of methylotrophs, 188, 364 from Methylobacterium extorquens AM1,188, 379 methanol dehydrogenase from Bacillus C1,188, 223 Hyphomicrobium X, 188, 203 Methylobacterium extorquens AM1, 188, 211 methylamine dehydrogenase from methylotrophic bacteria, 188, 242 Thiobacillus versutus, 188, 236 methylamine oxidase, 188, 227 muconate cycloisomerase, 188, 127 6-phosphogluconate dehydrogenase from Arthrobacter globiformis, 188, 343 Methylobacillus flagellaturn, 188, 337

Spin-echo experiments phospholipid hydroperoxide glutathione peroxidase, 186, 453 phthalate dioxygenase, 188, 62 proton release from yeast, 174, 600 serine-glyoxylate aminotransferase, 188, 362 serine hydroxymethyltransferases. 188, 366 transaldotase isoenzymes, 188, 406 trimethylamine dehydrogenase, 188, 251 triokinase, 188, 445 yeast long-chain alcohol dehydrogenase. 188, 172 yeast long-chain aldehyde dehydrogenase, 188, 176 Spectroscopy, see also specific techniques in analysis of biological chemiluminescence, 186, 600 aroxyl radical intermediates, 186, 351 Spermatozoa sea urchin nuclei, isolation, 170, 438 purification of histone subunits, 170, 460 Spherocytes Neurospora, handling, 174, 655 Spheroplasts yeast formation and lysis, 194, 654. 663 isolation, 194, 678 mitochondria, isolation, 194, 631 nuclear extract preparation, 194. 546 preparation for isolation of nuclei, 194, 739 plasmid chromatin, 170, 31, 37 RNA extraction, 180, 99 transformation, 194, 186, 263 Sphingomyelin reversed-phase HLPC, 172, 563 Spinal cord rat, substance P receptors, binding of anti-idiotypic antibodies, 178, 293 Spin diffusion reduction by random fractional deuteration, 177, 31 Spin-echo experiments homonuclear, lactate (2-CH) exchange, 176, 345 indirect detection of labels, 176, 347

Spin labeling Spin labeling proteins, 177, 86 Spin-spin coupling in quadrupolar nuclei NMR of ligandprotein complexes, 177, 319 Spin systems amino acids, identification computer analysis, 177, 155 long side-chain residues, 177, 139 methyl-containing residues, 177, 134 sequential assignments, 177, 155 short side-chain residues, 177, 135 identification by multiple-quantum NMR, 176, 133 proton, identification by heteronuclear NMR, 176, 108 Spin trapping in detection of superoxide and hydroxyl free radicals, 186, 127 free radicals, in correlation of radical formation with cell injury, 186, 611 Splenocytes fusion with myeloma cells, 182, 675 preparation for cell fusion, 184, 650 monoclonal antibody preparation, 182, 674 Splice junctions consensus sequence from different classes of organisms, 183, 256 identification, 183, 270 nonconforming sites, similarities, 183, 259 overview, 183, 252 3' sites, upstream signals, 183, 262 Spliceosomes assembly scheme, 181, 136 Splice signals assembly for control of expression of transferred genes, 185, 512 Splicing defects, nondirected mutations inducing, isolation, 181, 522 in Escherichia coli, genetic and molecular analyses, 181, 521 in expression of cDNA genes in mammalian cells, 185, 501 nascent transcripts, related structures, 180, 504

256 pre-mRNA analysis by transient transfection of HeLa cells, 181, 43 mammalian extracts active in, preparation and fractionation, 181, 3 nuclear miniextracts efficient in, preparation, 181, 20 SP6 in vitro, 181, 24 primer extension analysis, 181, 126, 141 self-, RNA by group I intervening sequence, 181, 558 solid-phase assessment, 181, 118 methods, 181, 109 in yeast active extract, preparation and fractionation, 181, 89 assay in vitro, 181, 92 efficiency, pre-mRNA sequence contribution, 181, 122 genetic analysis in mitochondria, 181, 539 Splicing complexes isolation and analysis, 184, 319 native gel electrophoresis, 180, 442 Spodoptera frugiperda cells, maintenance, 187, 497 5-1ipoxygenase expression with baculovirus vector, 187, 496 Sf9 cells-lytic baculovirus system construction, 182, 119 expression of mammalian proteins, 182, 117 recombinant viral genes and gene products, detection, 182, 124 Spores yeast enrichment, 194, 146 hydrophobic isolation, 194, 147 large-scale purification, 194, 110 random analysis, 194, 804 in genetic analysis, 194, 20 viability, ether test, 194, 146 Spore wall yeast biogenesis, defective mutants, isolation, 194, 121 large-scale purification, 194, 110

257 Sporulation in Bacillus subtilis, control by IPTG, 185,225 in Saccharomyces cerevisiae analysis in a/a and a/ct diploids, 194, 101 encoding genes, identification, 194, 110 monitoring, 194, 94 screening by ether test, 194, 146 specific genes or mutants interactions, 194, 109 screening and isolation, 194, 108 Spreading crosslinked DNA, 170, 158 nuclear, in visualization of synaptonemal complexes in yeast, 194, 110 Sputtering in FAB/MS and liquid secondary ion MS, 193, 203 SQ29548 in radioligand binding assays of thromboxane AJprostaglandin H2 receptors, 187, 402 SSREG.BAS program steady-state rate equation generation for enzyme and carrier-transport, 171, 164 Staining actin with fluorochrome-conjugated phalloidin, 194, 729 artery segments, 169, 62 avidin-biotin-peroxidase: detection of human T- and B-cell antigens, 184, 367 blots, 184, 425 counter-, see Counterstaining cytochemical Hansenula polymorpha, for localization of alcohol oxidase, 188, 414 catalase, 188, 413 procedural outline, 188, 412 developed embryos, for analysis of cartilage pattern, 190, 206 DNA for flow cytometry, 171, 579 fetal skeleton with alizarin red S, 190, 420 fluorescent, see Fluorescent staining

Staphylococcus

aureus

gels for I-D electrophoresis of nucleosomes, 170, 121 glycoproteins with PAS, 182, 536 thymol-sulfuric acid, 182, 536 immuno-, see Immunostaining mitochondria in cells of isolated nephron segments, 191, 258 negative, in immunoelectron microscopy of idiotype-anti-idiotype complexes, 178, 133 preelectrophoretic, proteins with fluorescent stains, 182, 480 proteins in electrophoretic gels with Coomassie Blue, 182,478 Coomassie Brilliant Blue R-250, 182, 437 copper, 182, 438 silver, 182, 438,482 on nitrocellulose replicas, 184, 433 quantitation, 182, 487 ribonucleoprotein, for visualization of yeast nucleolus, 194, 726 Schizosaccharomyces pombe nuclei, 194, 821 silver, see Silver staining sodium dodecyl sulfate gels for 2-D electrophoresis of DNA, 170, 132 SV40 minichromosomes or nucleosomes for electron microscopy, 170, 173 tissue sections after in situ hybridization of retinoid-binding protein mRNA, 189, 294 Triton-acid-urea gels for 2-D electrophoresis of proteins, 170, 137 vital, yeast vacuoles, 194,646 yeast bud scars and cell wall chitin with Calcofluor, 194, 732 yeast nucleolus with bismuth, 194, 723 yeast nucleus, 194, 99 Staphylococcus aureus

-antigen-antibody complexes, formation, 182, 695 and cell extracts, preadsorption, 182, 694 nonspecific binding of radiolabeled material, reduction, 182, 696 in vesicle immunoadsorption, 191, 839

Staphylococcus aureus ix-toxin Staphylococcus aureus a-toxin

permeabilization of mammalian cells to macromolecules, 171,867 Starch(bacterial glycogen) synthase tryptic peptides, liquid secondary ion MS analysis, 193, 483 Statistical analysis molecular phylogenies bootstrap method, 183, 657 character state approach, 183, 645 distance matrix approach, 183, 649 multiple sequences, 183, 442 nucleotide sequence branching order and dates, estimation, 183, 550 platelet interaction with artificial surfaces, 169, 113 proteins coding regions in genomic sequences, 183, 164 sequences significant pattern identification, 183, 388 similarities, significance, 183, 474 sequences divergence, 183, 531 statistical geometry on sequence space, 183, 505 splice junctions, branch point sites, and exons, 183, 252 Stearate disposition on stimulation of phosphatidylinositol breakdown, 191,688 formation from diacylglycerol, inhibition by RHC 80267, 191, 690 Stelar tissue ion transport mechanisms, 174, 287 Stellate cells rat liver identification and characterization, 190, 70 isolation and cultivation, 190, 58 purification from nonparenchymal cell isolates, 190, 55 role in retinoid metabolism, 190, 60 STEMS program extension for detection of pseudoknots and hairpins in RNA, 183, 307 Stereology renal epithelia, basic parameters, 191, 281

258 Sterilization affinity chromatography gels and columns, 182, 369 Steroid receptors family, identification of retinoid receptors as members, 189, 223 Steroids -protein conjugates, preparation, llN, 487 Stilbene disulfonates binding to band 3 protein, fluorescent assay, 173, 510 synthesis, 173, 509 Stimulus-secretion coupling analysis with isolated brush border membranes, 191, 675 in epithelial cells, models and specific aspects, 191,661 Stomach chief cells, isolation of peptic granules, 192, 165 glands acid formation, 191, 726 basolateral surface, perfusion, 192, 42 effects of histamine, carbacholine, and gastrin, 191, 728 isolated, intracellular potential and resistance measurements, 192, 90 isolation, 191, 645, 724 loading with Fura-2, 192, 63 membrane preparation, 192, 155 parietal cells in, permeabilization, 192, 110 pepsinogen secretion in vitro, 192, 124 permeable, preparation and effects of inhibitors, 191, 731 preparation for analysis of pepsinogen secretion, 192, 125 rabbit, preparation, 192, 95 responsiveness to inhibitors, 191, 729 isolated cells purification, 171, 450 viability assessment, 171, 456 isolated H+,K+-ATPase-containing membranes, purification, 191, 734 mucosa antral, HCO~ secretion in vitro, 192, 141

259 canine, endocrine cells isolation and primary culture, 192, 176 peptide release studies, 192, 184 dissociation into individual cells, 171, 445 fundic, acid-inhibited, HCO3 secretion in vitro, 192, 141 intact, intracellular potential and resistance measurements, 192, 86 ion transport, electrophysiological studies, 192, 82 isolated effects of cimetidine and omeprazole, 191, 724 preparation, 191, 723 responsiveness to secretagogues, 191, 723 isolated cells, intracellular potential and resistance, 192, 91 lumen-tubular cell pathway, resistance, 171, 624 perfusion, 191, 644 pieces, digestion, 192, 96 rabbit, vesicle preparation, 192, 153 transepithelial electrical measurements, 192, 83 parietal cells cyclic nucleotide-related measurements, 191, 649 functional correlation with cAMP metabolism, 191, 659 functional measurements, 191,647 intact, cAMP-dependent protein phosphorylation, 191, 654 isolation, 191, 640 purification, 191, 646 rabbit, purification of physalaeminrelated immunoreactive peptides, 168,450 reperfusion injury, role of oxygenderived free radicals, 186, 715 Streptavidin in affinity chromatography of replicating SV40 chromosomes, 170, 50 antibodies, preparation, 184, 435 -biotin in colorimetric detection of DNAsequencing bands, 184, 614

Strepolysin O in ELISA of herpes simplex virus, 184, 547 in isolation of splicing complexes, 184, 324 -biotinylated fluorescent microsphere. coating with biotinylated probe, 184, 355 coating of biotinylated fluorescent microspheres, 184, 355 colorimetric enzyme assay, 184, 218 complexes with biotinylated enzymes, 184, 181 conjugation to chemiluminescent marker ABEl-H, 184, 487 coupling to enzymes, 184, 475 derivatives, affinity chromatography, 184, 171 enzymatic assays, 184, 208 -enzyme complexes, in detection of antigens on Western blots, 184, 437 -gold, in ultrastructural localization of retinal photoreceptor proteins, 184, 370 -horseradish peroxidase, in detection of biotinylated proteins, 184, 435 hydrazide derivatives, preparation, 184, 184 immobilization, 184, 634 immobilized in affinity purification of hormone receptors, 184, 636 interaction with biotinylated ligand, binding constant, 184, 633 in immunochemical detection of biotinylated proteins, 184, 435 isolation, 184, 80 native, preparation, 184, 83 nonspecific binding, comparison with avidin, 184, 206 oligotetramers, preparation, 184, 89 -peroxidase, in antigen detection, 184, 442 properties, 184, 55, 85 radioactive assays, 184, 208 truncated, preparation, 184, 86 Streptolysin O permeabilization of pancreatic acinar cells, 192, 285, 296

Strontium Strontium as Ca2+ analog in transport studies, 174, 80 Stylets aphid, in collection of sieve tube sap, 174, 305 Styryl dyes WW-638, raman spectrum, in determination of biomembrane surface potential, 171, 367 Subcloning general method, 182, 98 hybridomas, 182, 676 mutants produced by saturation mutagenesis, 185, 603,608 Subendothelium extracellular matrix coated surfaces, preparation, 169, 79 components, degradation by human platelets, 169, 85 megakaryocyte interaction with, 169, 87 platelet reactivity with, assay on coated dishes, 169, 82 in perfusion system, 169, 83 sulfate-labeled, preparation, 169, 86 human blood vessel, preparation, 169, 47 rabbit aorta, preparation, 169, 48 Subfractionation liver, 174, 54 Submitochondrial particles pH determination, 174, 153 Substance P anti-idiotypic antibodies biochemical and pharmacological properties, 178, 292 immunological characterization, 178, 283 production, 178, 276 consensus sequence, complementary peptide synthesis based on, 168, 21 idiotypic antibodies, production and selection, 178, 277 mRNA, differentially spliced, assay, 168, 634 Substance P receptors anti-idiotypic antibodies to, 178, 24 binding of anti-idiotypic antibodies, 178, 292

260 Substantia nigra ovine, oxytocin release, assay, 168, 199 Succinate buffer stock solutions, 182, 33 Succinimidyl 2,2,5,5-tetramethyl-3-pyrroline-l-oxyl-3-carboxylate spin labeling of protein amino groups, 177, 108 Succinylation egg-white avidin, 184, 383 Sucrose 14C-labeled, uptake by phloem, 174, 289 density gradient fractionation, in preparation of clone libraries in YAC vectors, 194, 261 gradients analysis, in cellular retinoid-binding assays, 190, 151 in analysis of nucleosome structure, 170, 55 loading into leaves, 174, 293 UDPG-dependent vectorial synthesis in sugarcane vacuoles, assay, 174, 563 Sugarcane vacuoles, UDPG-dependent vectorial sucrose synthesis, assay, 174, 563 Sugars altered, gas chromatography, 186, 519 apurinic/apyrimidinic sites, determination, 186, 518 derived Bn ions, analysis, 193, 621 ions in mass spectra of trimethylsilyl derivatives of nucleosides, 193, 833 moieties in antibodies, biotinylation, 184. 162 mucin-type chains, quantitative conversion to radioactive oligosaccharides, 179, 261 2-thiobarbituric acid assay, 186, 518 transport in erythroctyes biochemical analysis, 173, 234, 249 optical analysis involving osmotic swelling or shrinking, 173, 235 radioactive analysis, 173, 243 inhibition, 191, 773 in K-562 cells, 191, 772

261 Na+-independent, in intestinal epithelial cells, 192, 334 in normal and mutant yeast, 174, 617 uptake by Chlorella, analysis, 174, 390 Sugar transporter apparent 'affinity of p-azidobenzylphloretin and p-azidobenzylphlorizin, 191,771 Sulfate depleted media, proteoglycan undersulfation by cells in, 179, 428 labeled subendothelial extracellular matrix, preparation, 169, 86 production from cysteine by cells, 179, 434 in proteins or peptides assay, 193, 487 detection, 193, 482 Sulfation assay, 179, 431 microsomal, proteochondroitin, chondroitin, and chondroitin oligosaccharides, 179, 422 tyrosine in proteins expressed in mammalian cells, 185, 592 under-, proteoglycans by cells in chlorate-containing media, 179, 429 sulfate-depleted media, 179, 428 Sulfatoglycosphingolipids structure, biological sources, and properties, 179, 169 Sulfhydryl groups directed acid-activated H+,K+-ATPase inhibitors, effects on H ÷ transport and ATPase, 191,734 Sulfidoleukotrienes analysis, 187, 575 Sulfidopeptide leukotriene receptors binding assays, 187, 421 Sutfinic acid acid-butanol extraction, 186, 141 diazonium coupling reaction, 186, 144 Sulfobromophthalein binding to translocase, assay, 174, 51 Sulfolane mass spectra, 193, 881 Sulfur centered radical cations, chemical properties, 186, 178

Superoxide dismutase centered free radicals chemical properties, 186, 174 generation, 186, 169 identification, 186, 172 Sulfuric acid -thymol, staining of glycoproteins, 182, 536 Sulfurous acid -fuchsin, for yeast nucleolar staining, 194, 728 Supercritical fluid chromatography -MS analysis, 193, 726 Supeffusion in assay of prolactin secretion, 168, 254 isolated glomeruli as system for, 191, 136 isolated secretory coils of sweat gland, 192, 595 Superoxide dismutase amino acid sequence phylogeny, 183, 622 assay by chemiluminescence of luciferin analog, 186, 227 hematoxylin autoxidation, 186, 220 NAD(P)H oxidation-based chemical system, 186, 209 in blood, automated assay, 186, 232 Cu,Zndifferentiation from Mn-superoxide dismutase, 186, 229 erythrocyte assays, 179, 573; 186, 232 glycation, 179, 570 purification, 179, 573 deficient bacterial mutants in assay of superoxide radical role in mutagenicity, 186, 647 elevated mutagenesis, 186, 646 effects on endotoxin-induced circulatory shock, 186, 661 endotoxin-induced disseminated intravascular coagulation, 186, 657 exocellular, purification, 186, 249 extracellular in plasma, automated assay, 186, 232 in tissue homogenates and extracellular fluids, analysis, 186, 260

Superoxide dismutase ~H NMR spectrum, 177, 249 human, export from Escherichia coli, 185, 183 induction by synthetic dyes in Escherichia coli, 186, 641,644 inhibition, pH effects, 186, 224 mimic derived from desferrioxamine and

MnO2 preparation, 186, 243 properties, 186, 244 Mn-, differentiation from Cu,Zn-superoxide dismutase, 186, 229 role in myocardial preservation during heart transplantation, 186, 742 in whole bacterial cells, assay, 186, 237 Superoxide radicals chemiluminescence assay based on luciferin analogs, 186, 585 generating systems, hydroxyl radical production, 186, 17 generation by human granulocytes or monocytes, assay, 186, 586 photooxidation, 186, 637 subcellular fractions, assay, 186, 588 kinetics and mechanisms, 186, 95 oxidation of tocopherols, 186, 200 production by neutrophils and other phagocytic cells, kinetic microplate assay, 186, 567 reaction with hemoglobin, 186, 266 metal-polyaminocarboxylate complexes, 186, 117 role in human disease, 186, 5 mutagenicity, assay with bacterial mutants, 186, 647 spin trapping, 186, 127 Surface charge membrane, effects in analysis of ionic permeation mechanisms, 171, 700 Surface potential artificial phospholipid membranes, determination, 171, 342 biomembranes determination, 171, 364 effects on protein function, 171, 387

262 transport parameters in yeast, 174, 581 lipid coumarin dye as probe, 171, 376 yeast, effect on ion uptake, 174, 609 Surface tension detergent solutions, calculation, 172, 536 Surfactants nonionic, solubilization of membrane proteins, 172, 519 Surgery cell, for obtaining Chara internodal cells, 174, 445 for ex oivo studies of platelet adhesion to polymeric surfaces, canine model, 169, 95 micro-, see Microsurgery pituitary gland, 168, 236 Survival irradiated mice treated with radioprotectors, 186, 694 mammalian cells treated by heat and thiols, assay, 186, 701 Suspension cultures hepatocytes, 173, 567 hexose transport, 173, 616 liver cells, total, differential centrifugation, 190, 65 Mg~+ transport assay, 173, 549 neurons, transplantation, 168, 251 neutrophils, 187, 593 pH measurement, 172, 87 platelets preparation, 169, 119 stirred aggregation in, 169, 136 hemacytometer and electronic particle counting, 169, 145 washed, preparation and handling, 169, 458 protoplast isolation, 181, 152 red cells, reflection coefficient measurement, 173, 213 requirements for gradient centrifugation, 171, 465 single-cell, requirement for flow cytometry, 171, 564 whole cell amino acid loading, 174, 157 ouabain binding, 173, 683

263

Temperature

yeast, density, effect on transport parameters, 174, 584 Sweat visualization in vivo, 192, 583 Sweat gland dissociated cells, preparation, 192, 598 eccrine, functional analysis, 192, 583 SWISS-PROT database in EMBL data library, 183, 26 Symplast measurements in, 174, 287 Synapsin I antibodies IgG, immunoprecipitation of bovine synaptic vesicles, 174, 13 Synaptonemal complexes yeast, visualization, 194, 110 Synaptosomes calcium transported in, assay, 174, 3 mammalian brain, purification of ATPdependent Ca2÷ pumps, 172, 35

T Talin, see Thaumatin Tandem mass spectrometry derivatized glycosphingolipids, 193, 759 doubly charged tryptic fragments, 193, 426 electrospray, peptides, 193, 178 fast atom bombardment, peptides, 193, 174 general concepts, 193, 156 glycoconjugates: structure of oligosaccharide residues, 193, 607 glycolipids, 193, 738 reduced and permethylated compounds, 193, 753 underivatized compounds, 193, 744 hexapole and octapole devices, 193, 169 and high-energy collision-induced dissociation, in peptide sequencing, 193, 455 hybrid sector-quadrupole biochemical applications, 193, 192 instrumentation, 193, 180 mass reference standards, 193, 312 matrix effect, 193, 211 multisector magnetic instruments, 193, 131

negative ion, glycolipids, 193, 748 oligosaccharides, 179, 157 positive ion calibration mixture, 193, 873 glycolipids, 193,744 in protein sequencing, 193, 464 rf-only quadrupole reaction cells, 193, 159 in sequencing of phosphorylated and sulfated peptides, 193, 493 triple quadrupole biochemical applications, 193, 174 instrumentation, 193, 170 Target analysis in determination of molecular mass of membrane components, 172, 410 Taurocholic acid photoaffinity labeling derivative, synthesis, 174, 26 T cells cytolytic, in assay of retinoid immunomodulatory activity, 190, 254 epitopes identification, 178, 611 mapping by k-tuple frequency analysis, 183, 237 structure, amphipathic helix hypothesis, 178, 616 synthetic, as immunogens to induce antibodies to hepatitis B components, 178, 635 in vaccine construction, 178, 611 helper epitopes, addition to B-cell epitope peptides, 178,661 function, priming by synthetic pre-S(1) T epitope, 178, 649 in oioo activity, priming by synthetic HBcAg peptides, 178, 652 human, antigens, immunocytochemical detection, 184, 363 tumors involving, treatment with antiidiotype antibodies, 178, 341 Television camera in measurement of fluorescence, 192, 43 Temperature critical micelle, see Critical micelle temperature

Temperature effects on amino acid transport in red cells, 173, 156 detergent properties, 182, 257 phenylglyoxal modification of anion transport system, 173, 473 low, rat liver cells at, isolation, 190, 54 regulated yeast promoters applications, 185, 365 construction, 185, 357 testing, 185, 362 regulatory effects, superimposition on yeast promoters, 185, 351 related relaxation method for analysis of ion carriers in planar bilayers, 171, 280 Temperature-jump methods in analysis of RNA melting behavior, 180, 322 Teratogenesis retinoids, evaluation with disposition data, 190, 406 Teratogenicity retinoids correlation with transplacental and maternal pharmacokinetics during organogenesis, 190, 437 mechanistic studies, 190, 418 testing with micromass limb bud cell culture, 190, 427 in vivo, 190, 433 Teratology embryonic retinoic acid derived from maternally administered retinol, 190, 319 Terbium phosphorescence, in determination of biomembrane surface potential, 171, 372 Terminal maturases for rRNA, substrates and assays, 181, 369 Testis bovine, purification of phosphomannosyl receptors, 179, 306 1,3,4,6-Tetrachloro-3a,6a-diphenylglycoluril, see Iodogen 1,3,4,6-Tetrachloro-3c~,6a-glycouril, see Iodogen

264 12-O-Tetradecanoylphorbol 13-acetate effects in skin, modulation by retinoids, 190, 346 Tetraglycosylceramides perbenzoylation, 172, 544 Tetrahyrnena thermophila

rRNA precursor intervening sequence end labeling, 181, 562 in self-splicing and cleavage of RNA, 181, 558 Tetramethylammonium chloride oligonucleotide hybridization in, 182, 748 2,2,6,6-Tetramethyl-3-(bromoacetamido)piperidine-l-oxyl radicals spin labeling of proteins, 177, 118 2,2,6,6-Tetramethyl-3-(bromoacetamido)pyrrolidine-1-oxyl radicals spin labeling of proteins, 177, 118 N-(2,2,5,5-Tetramethyl-3-carbonylpyrrolidine- l-oxyl)imidazole spin labeling of proteins, 177, 114 N-(2,2,5,5-Tetramethyl-3-carbonylpyrroline- l-oxyl)imidazole spin labeling of proteins, 177, 114 2,2,6,6-Tetramethylpiperidine-1-oxyl radicals derivatives, spin labeling of proteins, 177, 104 2,2,5,5-Tetramethylpyrrolidine-l-oxyl radicals derivatives, spin labeling of proteins, 177, 104 2,2,5,5-Tetramethylpyrroline-1-oxyl radicals derivatives, spin labeling of proteins, 177, 104 Tetraphenylphosphonium in determination of yeast membrane potential, 174, 611 tritiated, in measurement of plasma membrane potentials, 171, 779 TFASTA program DNA library searches, 183, 91 Thaumatin and vinculin, double fluorescence immunolabeling in embryonic fibroblasts, 184, 351 Thermodynamics alkali metal/proton exchange, force-fow analysis, 173, 340, 345

265 amino acid transport in red cells, 173, 156 coupled transport systems in vesicles, 191, 480 melting-related parameters for RNA, 180, 315 nonequilibrium, in analysis of biological transport, 171, 397 transport in isolated membrane vesicles, 172, 317 Thermolysin digestion of proteins for MS analysis, 193, 369 Thermometers 3~p, in chemical shift studies, 177, 308 Thermosensitivity double-stranded DNA, 180, 240 mammalian cells, thiol effects in vitro, 186, 696 Thiirane heteroatom analog of epoxyeicosatrienoic acids, preparation, 187, 363 Thin-layer chromatography arachidonate-containing phosphoglycerides, 187, 162 arachidonic acid metabolites, 187, 377 in assay of ceramide glycanase, 179, 479 potato arachidonate 5-1ipoxygenase, 187, 299 2,3-dinor-thromboxane B2, 187, 48 -FAB/MS, glycosphingolipids, 193, 725 high-performance in assay for endoglycoceramidase, 179, 489 lipids, 1911, 32 human anticarbohydrate antibodies, 179, 298 inositol phospholipids from platelets, 169, 422 lipids from erythrocytes, 173, 228 -liquid secondary ion MS, glycosphingolipids, 193, 725 -mass spectrometry, intact mixtures of glycosphingolipids, 193, 642 phloretin and phlorizin derivatives, 191, 765 platelet-activating factor, 187, 152 plates, glycosphingolipids on, cell-/tdhesion to, 179, 551

Thiyl radicals thymidine photoproducts, 186, 504 and 2D thin-layer electrophoresis, peptides, 182, 623 a-Thioalkyl radicals chemical properties, 186, 178 Thiobacillus versutus

purification of methylamine dehydrogenase, 188, 236 Thiobarbituric acid in assay of lipid peroxides in human body fluids. 186, 67 malonaldehyde, 186, 408 13-cis-retinoic acid inhibition of microsomal lipid peroxidation, 190, 282 sugar damage, 186, 518 Thiobenzyloxycarbonyl-L-lysinate based chromogenic substrate assay for oocyte tissue plasminogen activator. 168,424 l-Thioglycerol mass spectra, 193, 877 Thiols and disulfides, interconversion, 193, 375 drugs, thiyl radical metabolites. 186. 318 effect on mammalian cell thermosensitivity in vitro, 186, 696 in hemoglobin, oxidation. 186, 271 promoted cystine uptake, in vitro assay, 186, 707 protein, status during quinone metabolism, 186, 627 pumping, in generation of thiyl radicals. 186, 325 reaction with nitro compounds, 186, 325 Thiolutin inhibition of transcription in yeast, 194. 418 Thiolysis esters by thiol-derivatized antibody assay, 178, 565 substrate synthesis for, 178, 564 4-Thiouridine 5'-triphosphate synthesis, 180, 397 Thiyl radicals chemical properties, 186, 176 generation by peroxidases, 186, 319 thiol pumping, 186, 325

Thiyl radicals role in enzymatic reactions, 186, 328 radiation damage, 186, 329 Threonine spin system identification, 177, 135 Thrombi deposition on canine e x vioo polymeric shunt surfaces, 169, 104 platelet, formation, volume, and height, definitions, 169, 41 Thrombin effect on citrated platelet-rich plasma, 169, 131 treated endothelial cell monolayers, platelet adherence, 169, 74 Thromboembolization effect of shear flow, 169, 99 /3-Thromboglobulin antigens, identification and distribution, 169, 227 preparation from low-affinity platelet factor 4, 169, 227 Thrombospondin associated heparin-binding peptides, 169, 265 dark-field electron microscopy, 169, 326 electrophoretic transfer, 169, 265 identification by peptide fingerprinting, 169, 264 IgG reactive against, preparation, 169, 261 iodination, 169, 262 micropreparative and analytical isoelectric focusing, 169, 261 molecular, identification by electron microscopy, 169, 266 preparation and purification, 169, 256 radioimmunoassay, 169, 262 released, assays, 169, 268 secretion from human blood platelets, 169, 251 Thromboxane biosynthesis-related enzymes, nomenclature, 187, 5 metabolites, GC/MS assay, 187, 42 Thromboxane A2 synthesis by U937 cells, 187, 535 WI-38 cells, 187, 538

266 Thromboxane A2/prostaglandin H2 receptors antagonists binding studies in intact human platelets, 187, 410 platelet membranes, 187, 412 characteristics, 187, 406 radioligand binding assays, 187, 397 Thylakoids extract preparation, 182, 191 Thymidine 3H-labeled, in assay of retinoid effects on epidermal renewal, 190, 77 photoproducts, isolation and properties, 186, 504 sensitized photooxidation, mechanism, 186, 506 Thymidylate synthase encoding gene, in analysis of RNA splicing in E s c h e r i c h i a coli, 181, 523 Thymine glycol assay, 186, 515 Thymol -sulfuric acid, staining of glycoproteins, 182, 536 Thymus rat, isolation of lymphocytes, 173, 778 Thyroglobulin bovine, k-tuple frequency analysis, 183, 241 Thyroid hormone receptors family, identification of retinoid receptors as members, 189, 223 Thyroid peroxidase role in generation of thiyl radical metabolites of thiols, 186, 324 Thyroid-stimulating hormone receptors, s e e Thyrotropin receptors Thyrotropin receptors anti-idiotypic antibodies to, 178, 26 Thyrotropin-releasing hormone degrading enzymes, assays, 168, 365 and metabolites, assay, 168, 567 Thyrotropin-releasing hormone deamidase assay, 168, 369 Thyrotropin-releasing hormone pyroglutaminyl aminopeptidase, s e e 5-Oxoprolyl-peptidase

267 Tight junctions formation in airway epithelial cells, 192, 569 Time-domain data acquisition in 2D NMR, 176, 28 Time-resolved arrays in mass spectrometry, 193, 84 Tissue cultures cartilage pieces, 190,426 for herpes simplex virus, 184, 545 for immunoprecipitation of small RNPs, 180, 472 renal cell lines, 191, 426 Tissue inhibitor of metalloproteases recombinant, refolding, 185, 192 Tissues armadillo, M y c o b a c t e r i u m leprae-infected, purification of phenolic glycolipid I, 179, 227 cellular retinol- and retinoic acid-binding proteins, distribution, 189, 315 disaggregation in preparation of cells for flow cytometry, 171, 568 for electrophysiological assay of opioid peptides, preparation, 168, 116 exocrine, isolation of physiologically responsive secretory granules, 174, 162 extract preparation for retinoid isomerase assay, 189, 497 fetal, appropriate age for neural transplantation and dissection, 168, 249 fractions, retinoid synthesis from retinol and/3-carotene, 189, 470 for glutaminylpeptide cyclase assay, preparation, 168, 360 homogenates, analysis of extracellular superoxide dismutase, 186, 260 immunoassay samples, measurements, 182, 705 isolation of plasma membranes, 172, 18 lipid peroxide extraction, 186, 389 mammalian isolation of nuclei, 170, 281 purification of DNA, 170, 286 N e c t u r u s gall bladder epithelium, mounting, 192, 652 for peptidylglycine monooxygenase assay, preparation, 168, 353

Toluene dioxygenase plant, slicing, in isolation of vacuoles, 174, 554 rat N-acetylglucosaminyltransferase distribution, 179, 404, 406, 407 membrane-associated bile salt-independent retinyl ester hydrolase activities, 189, 459 preparation for assay of superoxide dismutase, 186, 228 for in situ hybridization histochemistry, preparation, 168, 767 vegetative, Dolichos biflorus, purification of lectins, 179, 344 Titration 2-anilinonaphthalene-6-sulfonic acidavidin complex, 184, 235 antisera to inhibin and synthetic peptide fragments, 168, 596 biotin, 184, 239 coulometric, chloride, 173, 62 yeast mRNA, 194, 539 TLC, see Thin-layer chromatography Tobacco leaves, protoplast isolation, 181, 153 suspension cultures, protoplast isolation, 181, 152 Tobacco hornworm midgut isolation, 192, 599 short-circuiting, 192, 603 c~-Tocopherol, see Vitamin E Tocopherols chemical and physical oxidation, 186, 198 enzymatic oxidation, 186, 200 and other nutrients in serum or plasma, simultaneous HPLC, 189, 155 photochemical oxidation, 186, 200 Tocopheroxyl radicals detection by ESR, 186, 197 Tocopheryl esters hydrolysis in biomembranes, fluorescence assay, 186, 355 Toluene hydroxylation by cumyl hydroperoxide, assay, 186, 275 Toluene dioxygenase assays, 188, 40

Toluene dioxygenase properties, 188, 45 purification from Pseudomonas putida F l, 188, 42 2-(p-Toluidinyl)naphthalene 6-sulfonate fluorescence, in determination of biomembrane surface potential, 171, 370 electrostatic potential adjacent to membranes, 171, 354 Tomato protein inhibitor I synthesis, induction by di- and triuronides, 179, 569 Tonoplasts algal, transport studies, 174, 378 Chara cell, ion fluxes, 174, 455 preparation, 174, 558 yeast, isolated vesicles, transport studies, 174, 517 Toxicity developmental, retinoids, evaluation with disposition data, 190, 406 a-Toxin permeabilization of pancreatic acinar ceils, 192, 285, 296 Trachea guinea pig, isolated, preparation, 187, 625 hamster, epithelium differentiation and keratin synthesis, role of retinoids, 190, 91 morphological alterations, 190, 91 preparation, 190, 91 vitamin A deficiency, keratin gene expression during, 190, 95 rabbit, epithelium, electrogenic and electroneutral ion transporter regulation, 192, 549 Tracing in electron microscopy for RNA mapping, 180, 254 retrograde, combination with in situ hybridization histochemistry, 168, 780 Transaldolase assay, 188, 406 crystallization, 182, 788 properties, 188, 410 purification from Arthrobacter PI, 188, 407 Candida utilis, 182, 788

268 Transcription cell-free extracts for efficiencies, 170, 355 preparation, 170, 349 chromatin active in, nuclease digestion, 170, 317 chromatin template permitting, reconstitution, 170, 581 in Escherichia coli, optimization, 182, 93 heterologous genes in yeast, termination or processing signals, 185, 426 initiation sites in rDNA unit, mapping, 180, 103 in isolated nuclei, in pulse labeling of hnRNA, 180, 86 neuroendocrine peptide genes, assay, 168, 622 neuropeptide genes in nervous system tissue, assay, 168, 681 nuclear runoff assay in vitro, 190, 137 nuclei active in, preparation, 181, 30 plasmid DNA templates for, 180, 52 preRNA, 180, 455 reactions for small RNA synthesis, 180, 56 regulation by retinoid-receptor complex, assay, 189, 229 RNA polymerase III-based, analysis in vitro, 170, 347 RNA probes for high-resolution in situ hybridization histochemistry, 168, 772 SP6 pre-mRNA in vitro, 181, 24 by SP6/T7 system: synthesis of long capped transcripts in vitro, 180, 42 synthetic DNA templates for, 180, 52 termination, in expression of cDNA genes in mammalian cells, 185, 502 tRNA genes assay, 181, 385 reaction conditions, 181, 443 yeast extracts for, 181, 439 viral, in synthesis of capped mRNA in vitro, 180, 166 in yeast inhibition with 1,10-phenanthroline, 194, 418 temperature-sensitive RNA polymerase II mutants, 194, 419 thiolutin, 194, 418

269

Translocation RNA polymerase II-initiated, assay in vitro, 194, 545 in vitro

DNA encoding secretory proteins, 194, 675 for reverse biochemical protein synthesis, 194, 524 Transcription cassettes for cloned yeast genes, requirements, 194, 373 Transcription factors -nucleosome core complex, reconstitution, 170, 583 TFIIIA, purification from immature oocytes, 170, 357 Transection pituitary stalk, 168, 243 Transesterification lipid peroxides to methyl esters, 186, 390 pentafluorobenzyl esters, 186, 391 Transfection co-, see Cotransfection COS-1 cells, mediation by DEAE-dextran, 185, 507 expression vector for chimeric antifibrin antibody with t-PA activity, 178, 523 keratinocytes with keratin genes, 190, 29 mammalian cells, mediation by DEAE-dextran, 185, 530 electric fields, 185, 531 lipids, 185, 533 polybrene, 185, 532 red blood cells, 185, 534 murine teratocarcinoma cells, 190, 139 in production of chimeric antibodies, 178, 462 recombinant proteins in mammalian cells, optimization, 185, 567 protoplasts, 181, 153 secretory cells with insulin gene, 192, 227 S19 cells with lytic baculovirus plasmid DNA, 182, 121 teratocarcinoma F9 cells and differentiated derivatives relative efficiencies, 190, 116 transient techniques, 190, 115

transient in assay of retinoid gene activation capacity, 189, 267 HeLa cells for analysis of splicing, 181, 43 Transfectomas for production of chimeric antibodies, analysis and isolation, 178, 469 Transferrin one-step immunoaffinity purification, 184, 301 role in iron metabolism, 186, 32 Transforming growth factor-/3 biotinylation, 184, 279 chromatography, 184, 280 Transforming growth factor-fl receptors flow cytometric detection, 184, 281 Transfusion preparation of labeled rabbit platelets for, 169, 468 Transglutaminase, see Protein-glutamine yglutamyltransferase Transition metals effects on hematoxylin autoxidation. 186, 223 metabolism, 186, 29 Transit times epidermal, effects of retinoids, 190, 78 Translation amino-terminal processing during, 185, 398 cloned genes, optimization in Escherichia coil, 182, 103 efficiency, enhancement with two-cistron expression system, 185, 94 in expression of cDNA genes in mammalian cells, 185, 503 consensus sequence requirements. 185, 504 eIF-2-mediated efficiency, 185, 504 foreign genes in Escherichia coli, enhancing vector, 185, 115 high-level initiation, 185, 89 preprogastrin-releasing peptides, 168, 677 in yeast, sequence and structural requirements, 185, 366 Translocation protons by H+-ATPase in N e u r o s p o r a crassa, assay, 174, 668,674

Transplantation Transplantation heart, myocardial preservation during, role of oxygen scavengers, 186, 742 neural, in hypothalamus, 168, 247 organ preservation for, effect of ischemia-reoxygenation injury, 186, 81 pituitary, 168, 242 renal, free radical-mediated reperfusion injury during, assay, 186, 748 Transport active, binding energy and coupling rules for, 171, 145 algal systems, overview, 174, 366 analysis with cultured bovine corneal endothelial cells, 192, 571 biological analysis with perfused liver, 192, 485 nonequilibrium thermodynamics, 171, 397 in biomembranes carrier-linked, kinetic criteria, 171, 14 experimental paradigms, 171, 24 inhibition kinetics of carrier systems, 171, 113 kinetics, 171, 23 machinery, overview, 171, 3 models, 171, 33 system identification, isolation, and characterization, 171, 12 bryophyte systems, overview, 174, 366 carrier-mediated, steady-state rate equations, generation, 171, 164 continuous monitoring in cells and vesicles, fluorescence methods, 172, 122 coupled, systems in vesicles stoichiometry, 191, 479 thermodynamics, 191, 480 electrogenic sugars by Chlorella, 174, 396 systems, kinetic properties, 171, 731 kinetics, computer analysis, 172, 262 and metabolism, measurements in cultured cells, 173, 714 optical studies, 191, 469 rate measurement, principles, 173, 617 saturable, characteristics, 174, 572 theoretical equations, 174, 184

270 Transporters endocytotic removal and exocytotic insertion: regulation of membrane transport, 172, 49 Transposons bacterial, for shuttle mutagenesis in yeast, 194, 329 yeast, Ty, induced mutations in Saccharomyces cerevisiae, 194, 342 assay, 194, 348 chromosomal manipulations, 194, 356 cloning, 194, 357 marked insertions and portable genetic markers, 194, 357 Trehalose lipooligosaccharides containing, purification from mycobacteria, 179, 225 Tretinoin, see all-trans-Retinoic acid TRH, see Thyrotropin-releasing hormone Triacylglycerol disposition on stimulation of phosphatidylinositol breakdown, 191, 688 Trichloroacetic acid and deoxycholate, precipitation of proteins, 1112, 593 precipitable material, [2J4C]uridine incorporation, 182, 221 Trifluoroacetic acid neutral sugar hydrolysis in, 179, 80 Trifluoroacetylation for mass spectrometry, 193, 326 p-Trifluoromethoxyphenylhydrazone inhibition of vesicular glutamate uptake, 174, 21 3-(Trifluoromethyl)-3-aryl-3H-diazirines chemical properties, 172, 645 synthesis, 172, 644 3-(Trifluoromethyl)-3-(m-[125I]iodophenyl3H-diazirine synthesis, 172, 648 Trigalacturonic acid enzymatic digestion, 179, 567 isolation, 179, 568 proteinase inhibitor, 179, 569 A4,5-Trigalacturonic acid enzymatic digestion, 179, 567 isolation, 179, 568 proteinase inhibitor, 179, 569

271 Triglycosylceramides perbenzoylation, 172, 544 2',4',6'-Trihydroxy-4-nitro-dihydrochalcone, see 4-Nitrophloretin

Triiodothyronine effects on mitochondria, 174, 127 Trimethylamine dehydrogenase assay, 1811,251 properties, 188, 257 purification from bacterium W3A1, 188, 253 2-Trimethylammonium acetyl derivatives N-terminal, formation, 193, 478 Trimethylguanosine antibodies, in RNA protection experiments, 180, 465 2,2,7-Trimethylguanosine antibodies in affinity chromatography of nucleoplasmic snRNPs, 181, 241 immunoprecipitation of snRNPs in nuclear extracts, 181, 83 purification, 181,236 in purification of snRNP particles, 181, 232 Trimethylsilylation DNA and nucleoprotein hydrolysates, 186, 533 for mass spectrometry, 193, 327 nucleic acid components, 193, 791 Trinitrophenylation monoclonal antibodies, 184, 347 Trinucleotides in RNA branches, core composition, 181, 187 Triokinase assay, 188, 445 properties, 188, 450 purification from Candida boidinii KDI, 188, 448 Triphenylmethylphosphonium in measurement of red cell membrane potentials, 173, 94 tritiated, in measurement of plasma membrane potentials, 171, 779 Tris buffer, see Tromethamine Tris(hydroxymethyl)aminomethane, see Tromethamine

Trypsin Tris(perfluoroheptyl)-s-triazine reference ion masses, 193, 870 Triton shells membrane skeletal proteins, isolation, 173, 389 Triton X-100 -acid-urea gels for electrophoresis of core histones, preparation, 170, 544 2-D electrophoresis of proteins preparation, 170, 136 staining, 170, 137 -membrane protein, sedimentation coefficient and partial specific volume, 172, 700 Triton X-114 phase partitioning of proteins from exocytic transport vesicles, 191, 821 tRNA guanylyltransferase assay, 181,451 partial purification from Drosophila, 181,454 yeast, 181, 453 properties, 181, 456 tRNA ligase assay, 181, 463 yeast, purification from Escherichia coli, 181,463 tRNA nucleotidyltransferase assay, 181, 435 properties, 181, 438 purification, 181, 436 tRNA splicing endonuclease Saccharomyces cerevisiae

assay, 181, 472 polypeptide components, 181,480 purification, 181,473 Xenopus laevis

assay, 181, 512 purification, 181, 515 Tromethamine buffer stock solutions, 182, 35 high-resolution discontinuous slab gels, preparation, 170, 556 Trypsin cleavage of anion transporter, 173, 424 cleaved prostaglandin endoperoxide synthase, reversed-phase HPLC, 187, 483

Trypsin digestion of 3H-reductively methylated histones, 170, 379 prostaglandin endoperoxide synthase, 187, 482 proteins for MS analysis, 193, 365 in preparation of resealable membranes depleted of cytosolic components, 173, 346 in release of cells from culture for flow cytometry, 171, 567 Trypsin inhibitors bovine pancreatic lysine-26 monoderivative, spectral analysis, 177, 101 spin-labeled, preparation and purification, 177, 104 Trypsinization T84 cells, 192, 388 Tryptophan in biotin-bindingproteins, homology around, 184, 101 in proteins, analysis, 182, 599 spin system identification, 177, 138 starvation of pATH-transformed yeast, induction of hybrid protein expression, 194, 486 Tubers potato, purification of arachidonate 5lipoxygenase, 187, 268 Tubules small, short-circuit current, determination, 192, 15 Tubulin purification, 182, 336 Tubulovesicles proteins in, analysis, 192, 163 Tulipa hybrids guard cell protoplasts, preparation, 174, 320 Tumor cells, see also specific tumor cells ascites, murine ATP-depleted, [A]i/[A]odependence on [Na+]o/[Na+]i, 173, 774 membrane potential, assay with carbocyanine dyes, 172, 95 transport: symport of Na ÷ with amino acids, 173, 771 bioenergetics and phospholipid metabolism, NMR monitoring, 177, 446

272 embryonal carcinoma available cell lines, 190, 150 cell markers, 189, 342 culture methods, 189, 338 cytosol, preparation, 190, 150 differentiation markers, 189, 344 in response to retinoids, 189, 337 intact, and extracts, retinoic acid and retinol metabolism, 189, 525 lacking cellular retinoic acid-binding protein, isolation, 190, 154 mRNA, analysis, 189, 346 retinoid-binding proteins, analysis, 190, 148 epidermoid carcinoma A431, protein kinase C, effect of GM3and derivatives, 179,536 erythroleukemia Friend fusion with hybrid reconstituted vesicles, 171, 844 plasma membrane, fusion-mediated implantation of exogenous protein, 171, 846 K-562 membrane perturbation and photoaffinity labeling, 191, 771 sugar transport, analysis, 191, 772 MEL, nucleus isolation, 181, 294 isolation of 40S hnRNP particles, 181, 294 free-flow electrophoresis, 171, 530 growth anchorage-dependent, inhibition by retinoids, 190, 101 anchorage-independent, inhibition by retinoids, 190, 107 HeLa, see HeLa cells hepatoma, extracts, interferences with CAT assay abolition by EDTA and heat, 168, 695 identification, 168, 693 HL60 differentiation assay, 190, 121 ganglioside-induced, into monocytes/macrophages and neutrophils, 179, 538

273 and growth, 190, 118 retinoic acid-induced, analysis, 190, 120 freezing, 190, 122 oxidized proteins, assay, 186, 575 preparation, 186, 576 protein retinoylation, 189, 233 reconstitution, 190, 122 human histiocytic lymphoma, U937, thromboxane A2 synthesis, 187, 535 intraceUular glutathione, assay, 186, 706 mastocytoma K21, nuclear extract preparation, 181, 76 murine leukotriene C4 synthase, characterization, 187, 306 propagation, 187, 309 P815, maintenance and labeling, 190, 256 myeloma, preparation for cell fusion, 184, 648 and fusion with spleen cells, 182, 675 ovarian carcinoma, thermosensitivity, thiol effects in vitro, 186, 698 retinoblastoma cell culture, 190, 141 interstitial retinol-binding protein mRNA, Northern blot analysis, 190, 147 retinoid-binding proteins, analysis, 190, 141 small cell lung carcinoma bombesin/gastrin-releasing peptide receptors, characterization, 168, 481 cytosolic Ca 2+, assay, 168, 490 squamous, specific markers, downregulation by retinoids, 190, 42 teratocarcinoma, F9 and differentiated derivatives, transient transfection, 190, 115 differentiation, 190, 112 maintenance, 190, 111 retinoic acid-regulated genes, expression assays, 190, 131 stem cells, and differentiated derivatives, relative transfection efficiency, 190, 116 transfection, 190, 139

Tyrosine kinase T-lymphoma, Jurkat, isolation of nuclei, 180, 86 Tumors, see also specific t u m o r s ascites, i n testing of drug and antitumor agent efficacy, 171,601 B-cell, murine, treatment with antiidiotypic antibody-drug conjugates, 178, 369 chemoprevention by retinoids, animal models, 190, 395 insulinomas, derived/3 cell lines, establishment and culture, 192, 223 lymphoid anti-idiotypic antibody therapy, 178, 341 idiotypic determinants, 178, 342 mammary, induction, 190, 399 murine papilloma epidermal, chemical induction. 190. 383 systemic treatment with retinoids, 190. 384 in testing of retinoid therapy of keratinization disorders, 190, 382 topical treatment with retinoids. 190. 392 related antigens, anti-idiotype antibodies against, 178, 15 Tungsten replication of proteins for dark-field electron microscopy, 169, 328 Turbidity lipid vesicle, measurement, 189, 380 Turbulence effects in continuous flow tube method for transport measurement, 173, 164 Tyrosine phosphorylation in angiotensin by pp60 Src, 193, 485 in proteins, spin labeling, 177, 114 spin system identification, 177, 138 sulfated peptides containing, liquid secondary ion MS analysis, 193, 486 sulfation in proteins expressed in mammalian cells, 185, 592 Tyrosine hydroxylase, see Tyrosine 3monooxygenase Tyrosine kinase PDGF receptor-associated, effects of GMI and GM3, 179, 523

Tyrosine kinase

274

pp60sr¢, phosphorylation of Tyr in angiotensin, 193, 485 Tyrosine monooxygenase transcripts, detection in cultured adrenal medulla, 168, 688 Tyrosine 3-monooxygenase -antibody reactions, analysis, 168, 379 antisera, preparation, 168, 376 in median eminence and superior cervical ganglion, immunoblot assay, 168, 371 purification, 168, 372

U U46619 in radioligand binding assays of thromboxane AJprostaglandin H2 receptors, 187, 404 Ubiquitin -protein conjugates degradation, 185, 390 formation, 185, 388 immunological detection in vivo, 185, 392 Ultracentrifugation in isolation of natural histone octamers, 170, 442 in sucrose gradient analysis of nucleosomes, 170, 70 Ultrafiltration in estimation of sulfobromophthalein binding to bilitranslocase, 174, 52 in protein concentration and solute removal, 182, 75 recombinant Fv and Fab fragments, 178, 510 Ultrafiltration coefficient capillary: assessment of glomerular filtration, 191, 115, 125 Ultramark 2500F reference ion masses, 193, 872 Ultrasonication leaf material, 174, 537 Ultraviolet light absorbance, in analysis of RNA hydrolysates, 193, 803 absorption, in assay of proteins, 182, 50 induction of crosslinking of nucleoprotein complexes, 170, 391

mutagenesis in yeast, 194, 278 protein crosslinking to preRNA, 181, 68 RNA crosslinking to cell monolayers, 180, 413 proteins in vivo, 180, 410 RNA, analysis: structure-function relationships, 180, 418 photochemical oxidation of tocopherols, 186, 200 Ultraviolet spectrophotometry in analysis of sucrose gradients, 170, 93 RNA, for determination of absorbance melting curves, 180, 304 Ultraviolet spectroscopy detergents, 182, 256 retinoic acid analogs, 189, 32, 38 Ultraviolet-visible absorbance detectors for HPLC of proteins, 182, 398 Uracil ~H-labeled, in pulse labeling of prerRNAs, 180, 99 M13 templates containing, in saturation mutagenesis, 185, 599 in radiolabeling of yeast RNA and phosphoproteins, 194, 427 in RNA, modification by hydrazine, 180, 374 substituted ssDNA, isolation from phage, 185, 606 Uranyl formate negative staining of idiotype-antiidiotype complexes, 178, 133 Urea -acid gels for first-dimension separation of RNA fingerprinting, 180, 143 fractionation of histone HI, preparation, 170, 535 in buffers for solubilization of membrane proteins, 182, 258 -Triton X-100-acid gels for electrophoresis of core histones, preparation, 170, 544 2-D electrophoresis of proteins preparation, 170, 136 staining, 170, 137 Uridine 14C-labeled, incorporation into trichloroacetic acid-precipitable material, 182, 221

275

Vacuoles

transport across red cell membranes, kinetics, 173, 260 Uridine 5'-diphosphoglucuronic acid preparation, 179, 275 Uridylylation Pit protein, effect on adenylyltransferase, 182,799 Uridylyltransferase regulation of glutamine synthetase formation, 182, 807 Urinary bladder mammalian, epithelium, physiological studies, 192, 632 pH and proton pumping, assay in cell culture, 174, 144 toad, model for transepithelial Na ÷ transport studies, 192, 683 Urine l l-dehydrothromboxane B2, assay, 187, 39 human malondialdehyde, HPLC assay, 186, 429 nucleoside fraction, trimethylsilyl derivatives, preparation, 193, 794 oligosaccharides, analysis and purification, 179, 55 8-hydroxy-2'-deoxyguanosine, HPLC with electrochemical detection, 186, 521 rat, malondialdehyde, HPLC assay, 186, 429 Ussing chambers in analysis of transepithelial electrolyte transport, 192, 361 edge leaks direct demonstration, 192, 698 minimization, 192, 707 quantification, 192, 700 UTase, see Uridylyltransferase UTP-agarose synthesis, 188, 340

V Vaccines idiotype, for poliovirus, canine parvovirus, and rabies virus, 178, 375 for infectious diseases, anti-idiotype antibodies as, 178, 7

peptide, based on peptlde epitopes with enhanced immunogenicity, 178, 659 synthetic, construction using T cell epitopes, 178, 611 Vaccinia virus mRNA capping enzyme applications, 181, 179 assay, 181, 172 purification, 181, 177 reactions, 181, 172 structure, 181, 171 purification, 181, 175 transcription, in synthesis of capped mRNA in vitro, 180, 168 vectors for heterologous gene expression in mammalian cells, 185, 490 Vacuoles Chara cell, contents, sampling, 174, 445 giant algal cells, perfusion, 174, 434 guard cell patch-clamp measurements, 174, 312 preparation, 174, 313 membranes, see Tonoplasts pH in living cells, measurement, 173, 745 plant isolation, 174, 553 transport assay, 174, 560 sugarcane, UDPG-dependent vectorial sucrose synthesis, assay, 174, 563 yeast fotation, 194, 655 fluorescence microscopy, 194, 645 immunofluorescence studies, 194, 601, 649 isolated arginine permease characterization, 174, 504 arginine transport, comparison with Neurospora, 174, 515 transport assay, 174, 510 marker enzyme assays, 194, 655 membrane vesicles, transport studies, 174, 517 preparation for transport studies, 174, 505 protein sorting to, fidelity, 194, 659 purification, 194, 653 purity assessment, 194, 657 yield, determination, 194,655 vital staining, 194, 646

V~ence Valence Mn, in superoxide dismutase mimic prepared from MnO2 and desferrioxamine, 186, 245 Valine spin system identification, 177, 134 Valinomycin in analysis of cation-anion transport: electrogenicity and electroneutrality, 173, 287 ion carrier in planar bilayers, relaxation and noise analyses, 171, 274 Vanadium grid staining in electron microscopy, 194, 626 Vanadyl -ribonucleoside complexes, in preparation of cytoplasm for RNA isolation, 180, 8 Vanadyladenosine preparation, 180, 416 Vaporization solvent, in formation of multilamellar vesicles, 171, 200 Vascularization neo-, see Neovascularization Vascular wall extracellular matrix, preparation, 169, 49 preparation, 169, 49 Vasoactive intestinal peptide purification from intestine, 168, 302 Vasodilation topical, in bioassay of arachidonic acid metabolites, 187, 382 Vasopressin affinity chromatography, 168, 585 analysis with monoclonal antibodies, 168, 574 cellular action, analysis, 191,551 conjugation to carrier protein, 168,576 effects on mitochondria, 174, 119 metabolites, assay, 168, 385 mRNA, in brain dot-blot analysis, 168, 407 nonradioactive visualization in hypothalamus, 168, 753 Northern blot analysis, 168, 405,407 preparation, 168, 400 in situ hybridization, 168, 409 solution hybridization assay, 168, 404

276 radiobinding assays, tracer effects, 168, 581 radiolabeling and tracer purification, 168, 576 Vasopressin-converting aminopeptidase assay, 168, 385 Vasoregulation pulmonary, role of eicosanoids, 187, 607 Vectorial processes coupled, binding energy and coupling rules for, utilization, 171, 145 Vectors for expression of cDNA genes in mammalian cells, 185, 495 heterologous genes in mammalian cells, 185, 487 with tac promoter, for transcription optimization in Escherichia coli, 182, 96 Vegetable oil suspension of retinoic acid in, 190, 234 Venom snake, preparation of botrocetin, 169, 152 Vesicles apical membrane isolation from renal epithelial cell lines, 191, 501 stimulated proteins in, analysis, 192, 163 size and orientation, 192, 162 ATP-dependent Ca2÷ pump reconstitution into, 172, 35 brush border membrane in analysis of transport systems, evaluation, 192, 415 functional, preparation, 192, 410 intestinal heterogeneity, relationship to tissue heterogeneity, 172, 355 perturbation and photolabeling by azides, 191, 767 transport studies, 172, 346 renal isolation, 191, 586 perturbation and photolabeling by azides, 191, 767 transport studies kinetics, 192, 422 measurement methods, 192, 413

277 for characterization and kinetic analysis of transport systems, 192, 409 coupled transport systems stoichiometry, 191, 479 thermodynamics, 191, 480 dimyristoylphosphatidylcholine, preparation, 171, 256 endocytotic, renal characterization, 191,512 isolation, 191, 509 exocytic transport analysis, 191, 820 formation, 191, 816 isolation, 191, 817 recovery from polarized MDCK cells in vitro, 191, 813 germinal, Xenopus laevis isolation, 181, 513 purification of tRNA splicing endonuclease, 181, 515 Golgi, rat liver glycoprotein sialate 7(9)-O-acetyltransferase, assay and properties, 179, 416 isolation, 182, 229 preparation and storage, 179, 418 hepatocyte membrane isolated, bile acid transport mechanisms, 192, 517 isolation, 192, 517 H+,K+-ATPase-containing, pH gradient formation, assay, 192, 161 hybrid, reconstituted Sendai virus envelope fusion with target cells, assay, 171, 844 fusogenic activity estimation, 171, 839 preparation, detergent selection for, 171, 842 immunoadsorption, in analysis of hepatocyte plasma membrane biogenesis, 191, 837 inside-out one-step ion transport measurements, 173, 371 pH effects on vesiculation, 173, 375 preparation from red cell membranes, 173, 368 stability, 173, 375

Vesicles red cell membrane, Na +,K+-pump stoichiometry and coupling, 173, 377 intestinal membrane basolateral Ca2+ uptake, 192, 448 heterogeneity, relationship to tissue heterogeneity, 172, 355 transport studies, 172, 346 CI- transport, 192, 405 isolated, electrophysiology, 192, 404 NaC1 transport pathways, 192, 389 Na" transport, 192, 399 lipid preparation, 171, 255 reconstitution of bacteriorhodopsin into, 171, 267 as retinoid carrier, preparation, 189, 404 retinoid exchange with rod outer segment membranes, 189, 402 size distribution in suspensions, 172, 383 lipid-protein, reconstituted, properties, 171, 263 membrane bulk D20 diffusion across, 173, 199 ion channel-mediated fluxes, assay, 172, 155 ion fluxes, measurement by rapidreaction methods, 172, 301 isolated nonhomogeneous populations, transport studies, 172, 325 size distribution, photon correlation spectroscopic study, 172, 330 tracer studies, 172, 313 partitioning in aqueous polymer twophase systems, 171, 547 renal proximal tubular heterogeneity, relationship to tissue heterogeneity, 172, 355 transport studies, 172, 346 membrane potential, optical measurement, 172, 102 multilamellar, preparation, 171, 198 nonhomogeneous populations, transport in, 172, 325 oligolamellar, preparation, 171, 208 ApH determination, 174, 131

Vesicles pH gradient measurements optical methods, 191, 471 principles, 191, 470 phosphatidylcholine donor, preparation, 173. 225 phospholipid ionic currents, estimation, 172, 341 preparation, 189, 374 turbidity measurement, 189, 380 planar membrane formation from, 171, 231 plasma membrane apical protein analysis, 191, 836 resolution, 191, 831 vesiculation, 191, 829 basolateral isolation, 192, 535 properties, 192, 537 protein analysis, 191, 836 resolution, 191, 831 vesiculation, 191, 829 canalicular isolation, 192, 535 properties, 192, 537 contralumenal isolation from kidney, 191, 450 purity, 191, 460 everted, isolation and transport function, 174, 667 glutathione transport studies, 173, 529, 532 hepatic, alanine transport, 174, 36 isolation from medullary thick ascending limb, 191, 461 renal inner medulla, 191, 465 lumenal isolation from kidney, 191, 450 purity, 191, 460 preparation from erythrocyte ghosts, 173, 514 rabbit gastric mucosa, 192, 153 proton electrochemical potential gradient in, 172, 63 reconstituted, pH determination, 174, 153 renal cortical, ATP-driven H + transport, 191, 505 retinol-phosphatidylcholine carrier, preparation, 1911, 15

278 sarcolemmal, cardiac, charge movements during Na+/Ca + exchange, 172, 155 seminal, ovine, isolation of prostaglandin endoperoxide synthase, 186, 432 separation, in determination of lipophilic ion distribution, 172, 69 size distribution determination by dynamic laser light scattering, 172, 364 intensity fluctuation spectroscopy, 172, 383 photon correlation spectroscopy, 172, 330 heterogeneity, effects on membrane potential, 172, 340 suspensions for planar bilayer formation, constraints, 171, 232 synaptic glutamate accumulation assay, 174, 11 characterization, 174, 16 preparation, 174, 13 response to glutamate analogs, 174, 22 toad bladder, Na ÷ transport, 192, 694 transcytotic, in hepatocyte couplets, analysis, 192, 509 transport in, continuous fluorescent monitoring, 172, 122 tubulo-, s e e Tubulovesicles unilamellar preformed, spontaneous insertion of integral membrane proteins, 171, 253 preparation, 171, 201 vacuolar membrane, transport studies in yeast, 174, 517 water permeability, optical measurement, 191, 562 Vesicular gland sheep, preparation of hgtl0 cDNA library, 187, 471 Vesicular stomatitis virus infection of MDCK cells, 191,815 Vesiculation spontaneous, in production of small unilamellar vesicles, 171, 204 Vibrating probes for ion flux measurements effects of probe mixing, 174, 468

279 one-dimensional probes, 174, 463 two-dimensional probes, 174, 467 for scanning extracellular current density application to epithelia, 171, 803 construction, 171, 796 data interpretation, 171, 809 principles, 171, 799 system integration and grounding, 171, 801 Vicia faba

guard cell protoplasts, preparation, 174, 318 Video methods measurement of volume and shape changes in isolated tubules, 191, 371 Vinculin and talin, double fluorescence immunolabeling in embryonic fibroblasts, 184, 351 Viruses, see also specific viruses associated network epitopes, 178, 8 -insect cell system for mammalian protein expression, 182, 117 mammalian, as transient expression vectors, 182, 117 transformation of/3 cells, 192, 227 Viscosity membrane, time-resolved fluorescence depolarization study, 172, 462 proteins, in determination of size, molecular weight, and presence of subunits, 182, 584 Visual cycle analysis by short-term incubation of isolated retinal pigment epithelial cells, 190, 3 retinoids bound to interstitial retinolbinding protein during, 189, 325 Vitamin A analysis in vivo, 190, 309 binding proteins cellular complexes with [3H]retinol, preparation, 189, 451 crystallization, 189, 519 gel electrophoresis, 189, 299 ligand interactions, 189, 514 mammalian, structure-function analyses, 189, 506 radioimmunoassay, 189, 276

Vitamin A separation of interstitial retinolbinding protein from, 189, 327 tissue distribution, 189, 315 crystal preparation, 189, 283 gene structure and expression, 189, 203 interstitial assay, 189, 330 cloning, 189, 212 mRNA from retinoblastoma cells, Northern blot analysis, 190, 147 purification, 189, 209 retinoids in, extraction, 189, 328 separation from cellular retinaldehyde- and retinol-binding proteins, 189, 327 sequences, 189, 212 tryptic peptides, characterization. 189, 210 Western blot analysis, 190, 144 parasite, retinol-ivermectin crosscompetition for, 189, 352 physicochemical properties, 189, 193 purification from plasma, 189, 193 for X-ray crystallography, 189, 282 radioimmunoassay, 189, 271 serum, gel electrophoresis, 189, 299 transfer of retinol to liposomes and across liposomal membranes, 189, 394 X-ray crystallography, 189, 281 capillary gas chromatography, 189, 99 and/3-carotene, retinoid synthesis from, assay and characteristics, 189, 470 carotenoids supplying, food composition tables for, assessment, 190, 237 deficient mice and rats, preparation, 190, 229 deficient subjects, immunotrophic methodology, 190, 259 deficient tracheal epithelium, keratin gene expression, 190, 95 depleted culture media, preparation, 190, 21 deuterated, administration, 189, 98 effect on lymphokine production and response, 190, 273 esterification by acyl-CoA-dependent pathway, assay. 189, 442, 446

Vitamin A lecithin-retinol acyltransferase, analysis, 189, 450 extraction from serum, 189, 98 fatty acyl esters, see Retinyl esters 3H-labeled physiological doses, preparation, 190, 305 in preparation of tritiated retinal, 189, 532 HPLC, 189, 50 isomers HPLC, 189, 73 reversed-phase separation, 189, 175 and ivermectin, cross-competition for parasite retinol-bindingprotein, 189, 352 kinetics in vivo, 190, 311 labeled chylomicrons, preparation, 190, 306 maternally administered, derived embryonic retinoic acid, assay, 190, 317 metabolism in cytosol, effects of microsomes, 189, 482 in epidermal cells: retinoic acid formation, 189, 530 by intact cells and cell extracts, 189, 525 microsomal metabolites, HPLC, 189, 485 and other nutrients in serum or plasma, simultaneous HPLC, 189, 155 -phosphatidylcholine carrier vesicles, preparation, 190, 15 preparation from retinal, 189, 78 pro-, see Provitamin A radioactive, analysis after isomerase assay, 189, 499 regulation of keratin expression, 190, 24 keratinocyte differentiation, 190, 18 and retinyl esters, separation, 189, 98 serum, liquid chromatographic assay, 189, 172 stable isotope dilution MS analysis, 189, 94 status in human population, biochemical and histological assessment, 19tl, 242

280 mass spectrometric assessment, 189, 94 total analysis, 189, 84 calculation in body, 189, 103 transfer from retinol-bindingprotein to liposomes and across liposomal membranes, 189, 394 turnover studies in vivo, 190, 308 all-trans-Vitamin A

3H-labeled, supply to retinal pigment epithelial cells in vitro, 190, 15 Vitamin A2 fatty acyl esters, see 3,4-Didehydroretinyl esters 3H-labeled, newly formed in epidermis, detection, 190, 212 synthesis by organ-cultured human skin, 190, 210 Vitamin E effect on endotoxin-induced disseminated intravascular coagulation, 186, 657 incorporation and distribution in biomembranes, fluorescence assay, 186, 355 serum, liquid chromatographic assay, 189, 172 Vitamins liquid chromatographic detection, 189, 179 Voltage clamping in control of membrane potential in giant algal cells, 174, 422 for impedance measurements in giant algal cells, 174, 427 in lepidopteran midgut, 192, 599 Volume cell, see Cell volume changes, microcomputer modeling, 172, 280 partial specific erythrocyte band 3 anion transporterTriton X-100 complex, 172, 700 membrane protein-detergent complexes, 172, 699 prostaglandin E~-Triton X-100 complex, 172, 703

281

Water

von Willebrand factor antibody titers, assay, 169, 161 bioassays, 169, 155 dark-field electron microscopy, 169, 326 multimer size, screening test, 169, 160 preparation from washed platelets, 169, 246 secreted from platelets, multimeric structure, 169, 249 secretion from platelets radioquantitative electrophoresis, 169, 248 release experiments, 169, 247

W Walmsley analysis thermodynamic effects on amino acid transport in red cells, 173, 158 Washing in isolation of human platelets from plasma, 169, 3 Water channels in membranes, identification, 191, 561 -dextran-polyethylene glycol, for partition of cells and organelles, 171, 532 diffusion across red cell membrane, 173, 192 D20, bulk diffusion across membranes, 173, 199 in erythrocytes, content determination, 173, 60 exchange half-time, absolute value and physiological meaning, 174, 193 flow in plants direct coupling to solute flow in cells, 174, 199 theory, 174, 184 flux, effect of p-azidobenzylphlorizin, 191, 776 intracellular levels in Amphiuma erythrocytes, 173, 333 ~80-labeled, exchange, in labeling of molecules, 193, 338 osmotic flow across red cell membrane, 173, 204

osmotic permeability of isolated renal tubule segments, measurement, 191. 249 passive permeability of plant cells, measurement, 174, 255,263 permeability of intact renal tubules, optical measurement, 191, 565 isolated vesicles, optical measurement, 191, 562 phospholipid membranes, effects of retinoids, 189, 375 reptilian distal tubule, 191, 221 permeability coefficient calculation with desk-top hybrid computers, 172, 274 relations of plant cells equations, 174, 186 pressure probe technique, 174. 191. 338 storage apoplast capacity, 174, 209 coupling with active process during CAM, 174, 207 structures in, geometry, relaxation matrix analysis, 176, 178 transmural flow in isolated perfused renal tubule segments, measurement, 191, 232 transport active transport theory, 174, 186 in avian renal tubules, measurement, 191, 226 coefficients, effects of unstirred layers, 174, 188 coupling with plant growth, 174, 211 equations, 174, 184 in gall bladder epithelium, electrophysiological studies, 192. 650 and plant mechanical properties, coupling, 174, 204 in plant tissues, mathematical model. 174, 189 in red blood cells and ghosts, measurement, 173, 160 in root, coupling to solute transport. 174, 214 across salivary acini, related ion fluxes, assay, 192, 33

Web 2086 WEB 2086 platelet-activating factor antagonist in vitro studies, 187, 458 in vivo studies, 187, 460 physicochemical properties, 187, 455 solution preparation and stability, 187, 456 WEB 2170 platelet-activating factor antagonist in vitro studies, 187, 458 in vivo studies, 187, 460 physicochemical properties, 187, 455 solution preparation and stability, 187, 456 Western blots antigen detection with streptavidinenzyme complexes, 184, 437 luminescent detection, 184, 442 Western blotting antibody detection, evaluation, 182, 668 in assay of hormone secretion from single cells, 168, 328, 336 carbohydrate receptors, 179, 118 in detection of protein degradation in intact cells, 186, 498 GAL4 protein from yeast, 185, 304 inhibin, 168, 610 interstitial retinol-binding protein, 190, 144 platelet PADGEM protein, 169, 319 thrombospondin peptides, 169, 265 Western immunoblot analysis proteins expressed in mammalian cells, 185, 595 Wheat germ isolation of deoxyribonucleoprotein, 170, 440 purification of exocellular superoxide dismutase, 186, 249 histone subunits, 170, 463 leaves mesophyll cells, isolation of deoxyribonucleoprotein, 170, 441 protoplasts, subcellular fractionation, 174, 526 Wing buds anterior margin, implantation of retinoic acid-impregnated bead, 190, 205

282 X Xanthine oxidase in biological tissues, assay, 186, 651 -hypoxanthine, in assay for superoxide dismutase, 186, 227 X e n o p u s laevis, see African clawed frog X-ray crystallography data collection, 189, 285 phase determination, 189, 285 retinol-binding proteins, 189, 281 Xylem electrophysiology, 174, 284 transport, perfusion studies, 174, 277 vessels measurements in, 174, 286 properties, 174, 280

Y Yeast, see also specific y e a s t s actin staining with fluorochrome-conjugated phalloidin, 194, 729 allele recovery, 194, 298 asci micromanipulation and dissection, 194, 21 wall digestion with snail juice, 194, 32 ascospores isolation of RNA, 194, 401 pure and viable, preparation, 194, 125 separation, 194, 34 binding of monovalent cation, 174, 610 cation composition, alteration, 174, 607 cell immobilization, 194, 576 cell isolation, 194, 37 cell structures, immunofluorescent detection, 194, 599 chromosomal footprinting, 194, 557 chromosome structure, analysis with H O endonuclease, 194, 143 CO2 levels, continuous estimation, 174, 602 conditional lethal, screening for defects in pre-tRNA processing, 181,401 culture media, 182, 155 culture vessel, 182, 156 CUP1 gene, isolation, 185, 321

283 DNA genomic bacterial transformation, 194, 323 preparation, 194, 322 sequencing, 194, 552 embedding for immunoelectron microscopy, 194, 611,614 ethanol dehydration, 194, 616 expression systems, overview, 185, 6 extract preparation, 182, 154 buffers for, selection, 182, 167 by enzymatic digestion of cell wall, 182, 164 with glass beads, 182, 162 nucleic acid removal, 182, 167 proteolysis during, prevention, 182, 168 fixation for immunoelectron microscopy, 194, 611,614 protocol, 194, 586 genes cloned expression vectors, 194, 373 propagation and expression with 2~m circle-based vectors, 185, 234 disruption, 194, 290 isolation with hgtl I system, 194, 23O mapping, 194, 38 replacement, 194, 293 growth conditions for immune mapping, 194, 614 growth and harvesting, 182, 154 heterologous gene expression with inducible copper metallothionein promoter, 185, 319 overview, 185, 231 high-efficiency expression vectors based on P G K promoter, 185, 329 with high permease activity, preparation, 174, 628 homothallic, conversion to heterothallism, 194, 144 as hosts for overexpression of foreign genes, 182, ll4 immunofluorescence studies, 194, 565 detection of specific structures, 194, 599

Yeast strain selection and growth conditions. 194, 569 integrative DNA transformation, 194, 281 intracellular ion distribution, 174, 608 ion transport determination, 174, 603 kinetics, 174, 606 isolation of deoxyribonucleoprotein, 170, 439 genomic DNA, 194, 552 poly(A)÷ RNA, 194, 402 lysis, 182, 159 mating types, switching, applications, 194, 132 membrane potential, determination with carbocyanines, 174, 615 glass microelectrodes and microfunnels, 174, 650 lipophilic anions, 174, 616 microelectrodes, 174, 611 quaternary phosphonium compounds, 174, 611 membrane surface potential, effect on transport parameters, 174, 581 methylotrophic, purification of formaldehyde dehydrogenase, 188, 456 formate dehydrogenase, 188, 460 microscopy, 194, 572 mitochondria function and assembly, 194, 627 isolation, 194, 631 preparation, 182, 173 protein import, 194, 629, 637 mutant, sugar transport, analysis, 174. 617 nuclei, preparation, 182, 171 pH determination, 174, 596 effect on cation uptake, 174, 608 phosphate-proton cotransport system, repression or derepression studies, 174, 608 photomicroscopy, 194, 572 plasmids bacterial transformation, 194, 323 preparation, 194, 322 polyploid strains, construction, 194, 138

Yeast protein synthesis in vitro reverse biochemical methods, 194, 520 translation system, 194, 536 proton extrusion, analysis, 174, 592 protoplasts, preparation, 174, 508 purification of histone subunits, 170, 459 Rec mutant, isolation, 194, 119 ribosomal core particles and proteins, preparation, 194, 469 ribosomal proteins functional analysis, 194, 467 structural analysis, 194, 453 ribosomes and ribosomal subunits, isolation, 194, 454 RNA low-molecular-weight, preparation and analysis, 194, 405 prepared, quality evaluation, 194, 404 secretion and modification of heterologous proteins, overview, 185, 232 shuttle plasmids in, recovery into Escherichia coli, 194, 319 small ribonucleoproteins, preparation and analysis, 194, 405 spheroplasts formation and lysis, 194, 654 transformation with YAC vectors, 194, 263 Spo- genes, cloning, 194, 123 spores, purification, 194, 124 spore walls mutants, isolation, 194, 121 purification, 194, 124 sporulation mutant spo13 in identification and analysis of Rec- mutations, 194, 117 segregation genes, 194, 120 recovery systems, 194, 111 storage, 182, 159 strains free of 2-/zm circles, isolation, 185, 258 isogenic, construction, 194, 136 manipulation, vectors for, 185, 269 specificity for structural and functional analysis, 194, 477 subcellular fractionation, 182, 171 sugar transport, analysis, 174, 617

284 surface potential, effect on ion uptake, 174, 609 suspensions density, effect on transport parameters, 174, 584 filtrates, organic acids in, estimation, 174, 602 transformation lithium-based method, 185, 283 with synthetic oligonucleotides, 194, 362 transport of amino acids and selected anions, 174, 623 transport processes in intact isolated vacuoles arginine permease characterization, 174, 504 arginine transport, 174, 515 measurement system, 174, 510 in isolated vacuolar membrane vesicles, analysis, 174, 517 kinetics, 174, 567 measurement techniques, 174, 587 ubiquitin-dependent proteolysis, detection and inhibition, 185, 387 ultrathin sections immunogold labeling, 194, 608 removal of epoxy resins, 194, 625 vacuoles flotation, 194, 655 fluorescence microscopy, 194, 645 immunofluorescence studies, 194, 649 marker enzyme assays, 194, 655 preparation for transport studies, 174, 505 protein sorting to, fidelity, 194, 659 purification, 194, 653 vital staining, 194, 646 vegetative cells, isolation of total RNA, 194, 399 Yeast artificial chromosomes vectors, clone libraries in, preparation, 194, 251 Yeast proteinase B enzymatic assay, 194, 447 in Golgi apparatus, assay, 194, 658 overlay test, 194, 435

285

Zuker-Stiegler algorithm

vacuolar biochemical analyses, 185, 379; 194, 444 genetic analyses, 185, 372; 194, 435 interference with yeast studies, prevention, 194, 428

Z Zea mays, see

Maize

Zeugmatin and integrin, double fluorescence immunolabeling in embryonic heart cells, 184, 350

Zinc intracellular quenching of quin2, 172, 251 Zinostatin -anti-DNA idiotypic antibody conjugates preparation, 178, 424 properties, 178,427 selective elimination of anti-DNA idiotype-positive human lymphocytes, 178, 428 Zuker-Stiegler algorithm and associated programs, RNA structure prediction, 180, 271

Contributor Index Boldface numerals indicate volume number.

A Abarztia, Patricio, 189, 337 Abe, Akiro, 179, 559 Abe, Yuki, 183, 3 Abel, Yvon, 183, 428 Abelson, John N., 180, 63; 181, 89, 97, 463, 471 Abildgaard, Frits, 176, 311 Abood, Leo G., 178, 212 Abraham, Nader G., 187, 372 Abrahms6n, Lars, 185, 144 Adam, Stephen A., 180, 410 Adamo, Sergio, 190, 81 Adams, Alison E. M., 194, 565, 729 Adams, Margie, 184, 343 Adler, Alice J., 189, 213 Aebi, Markus, 181, 43 Agrawal, S., 168, 753 Aguila, M. C., 168, 716 Aharony, David, 187, 414 Ahmed, Zahur, 192, 309 Ahn, Bong-whan, 186, 464 Akabas, Myles A., 191, 572 Akasaka, Kazuaki, 186, 157 Akerboom, Theo P. M., 173, 523 Al-Awqati, Qais, 172, 49, 85; 191, 253,572 Albertsson, Per-A.ke, 171, 532 Albright, C. F., 194, 682 Albright, George, 178, 49 Alexander, Stephen, 179, 505 Allaerts, Wilfried, 168, 47 Allen, Roberta, 168, 431 Allshire, Ashley, 174, 68 AI-Mallah, N. R., 189, 167 Altenberg, G., 192, 650 Altman, Russ B., 177, 218 Altman, Sidney, 181, 569

Alvarez, Javier, 173, 368 Amano, Junko, 179, 261 Ambudkar, Indu S., 192, 26 Ames, Bruce N., 186, 301,371,521 Amici, Adolfo, 186, 464 Amit, T., 178, 308 Ammann, Daniel, 172, 136 Amos, Brad, 190, 217 Amzel, L. Mario, 191, 371 Andersen, Olaf Sparre, 171, 62 Anderson, D. Eric, 177, 44 Anderson, Richard A., 173, 380 Anderson, Wayne F., 183, 438 Andersson, Eva, 190, 163 Andrews, P. C., 168, 72 Angel, Anne-Sophie, 193, 587 Anjaneyulu, P. S. R., 172, 609 Ansari, G. A. S., 186, 438 Anthony, Christopher, 188, 210, 216, 284, 298 Antoine, Catherine, 187, 82 Antoniades, Harry N., 169, 210 Anumula, Kalyan R., 193, 501 Anziano, Paul, 184, 328 Arai, H., 168, 753 Arai, Katsura, 179, 570 Archibald, F. S., 186, 237 Arciero, David M., 188, 89 Arfman, N., 188, 223,391 Argent, B. E., 192, 256 Argos, Patrick, 182, 751; 183, 352 Arion, William J., 174, 58 Aris, John P., 194, 735 Armstrong, William McD., 168, 144 Arnez, John G., 181, 569 Arps, Peggy J., 188, 386 Ashraf, W., 188, 21, 26 Asmus, Klaus-Dieter, 186, 168 287

288

CONTRIBUTOR INDEX

Asou, Hiroaki, 184, 405 Atencio, Edwin J., 183, 3 Attwood, Margaret M., 188, 314 Aubert, C., 189, 167 Ausiello, Dennis, 191, 551 Aust, Steven D., 186, 457 Avrameas, Stratis, 184, 469

B

Babbs, Charles F., 186, 137 Babel, Wolfgang, 188, 346, 350, 401,451 Bach, Montserrat, 181, 232 Bacon, David J., 183, 438 Baer, Madeline F., 181, 569 Bailey, John Stuart, 189, 356 Bakalova, Rumyana A., 186, 355 Baker, Peter F., 171, 817 Bala, Gregory A., 187, 226 Balaban, Robert S., 176, 330, 493 Balbas, Paulina, 185, 14 Baldi, I. M., 181, 510 Baldino, Frank, Jr., 168, 761 Baldwin, Stephen A., 174, 39 Ball, Mark D., 189, 446 Ballou, Clinton E., 185, 440 Ballou, David P., 188, 61 Banci, Lucia, 177, 246 Bangerter, Faan Wen, 189, 411 Banks, William A., 168, 652 Barasch, Jonathan, 172, 85 Barbone, Francis P., 194, 536 Bardin, C. Wayne, 168, 588 Barker, Winona C., 183, 31,333 Barkey, Ronnie J., 178, 308 Barnea, Ayalla, 168, 710 Barnett, Stanley F., 181, 293 Barnhouse, Steve, 183, 3 Barr, John R., 193, 501 Barry, Peter H., 171, 678 Bartles, James R., 191, 825 Barton, Geoffrey J., 183, 403 Barua, Arun B., 189, 136 Basus, Vladimir J., 177, 132 Batie, Christopher J., 188, 61 Batlle, Javier, 169, 244 Baulieu, Etienne-Emile, 184, 292 Baum, Bruce J., 192, 26 Baumert, H. G,, 172, 584

Baumgartner, Hans R., 169, 37 Bavykin, S. G., 170, 386 Bax, Ad, 176, 134, 151 Baxter, Robert C., 168, 309 Bayer, Edward A., 184, 5, 14, 49, 68, 80, 123, 138, 160, 174, 208, 217, 243,301, 340, 415,427,467, 560, 608, 618 Beall, L. Dawson, 184, 275 Beaucourt, J. P., 187, 70 Beck, Stephan, 184, 612 Becket, Daniel M., 194, 182 Bednarski, Mark D., 179, 275 Beilby, M. J., 174, 403 Beinfeld, Margery C., 168, 3 Belagaje, Rama M., 185, 94 Belfort, Marlene, 181, 521 Bell, Anne, 185, 351 Bellard, Maria, 170, 317 Bellomo, Giorgio, 186, 627 Belmonte, Elizabeth, 169, 371 Belyavsky, A. V., 170, 386 Bendich, Adrianne, 190, 252 Benenfeld, Bonnie J., 180, 130, 418 Ben-Hur, Haya, 184, 80, 217,415,427 Benjamin, Christopher W., 187, 535 Ben-Jonathan, Nira, 168, 144, 234 Bennett, Mark K., 191, 813 Benoit, Joseph N., 186, 729 Benson, Donelle M., 187, 520 Benton, David, 183, 3 Benveniste, Etty N., 178, 63 Benveniste, Jacques, 187, 125 Benz, R., 171, 274 Bereiter-Hahn, J., 186, 723 Berg, M., 168, 481 Berg, Trond, 190, 58 Berger, M., 186, 502 Berger, Melvin, 184, 619 Berger, Shelby L., 180, 3,325 Berglindh, Thomas, 192, 93 Berk, Arnold J., 180, 334 Berkowitz, Bruce A., 176, 330, 493 Berlin, Vivian, 194, 774 Berlove, David J., 178, 212 Berman, Cindy L., 169, 311 Bernlohr, David A., 189, 363 Bernstein, Paul S., 189, 494 Berteloot, A., 192, 409 Bertini, Ivano, 177, 246 Berzofsky, Jay A., 178, 611

CONTRIBUTOR INDEX Bessman, David, 169, 164 Better, Marc, 178, 476 Beyenbach, Klaus W., 191, 167 Beyer, Ann L., 180, 481 Beyer, Wayne F., Jr., 186, 242 Biber, J., 191, 494 Bicsak, Thomas A., 168, 422 Bielski, Benon H. J., 186, 108, ll3, ll6 Biemann, Klaus, 193, 295, 351,455, 886, 888 Bienkowski, Michael J., 187, 535 Biesalski, Hans K., 189, 181 Billeter, Martin, 177, 150 Billingsley, Melvin L., 184, 451 Birnboim, H. Chaim, 186, 550 Bishop, A. E., 168, 791 Bjerrum, Poul J., 173, 466 Blackburn, Carol C., 179, 542 Blackmore, P. F., 173, 534 Blair, Ian A., 187, 13 Blaisdell, B. Edwin, 183, 388 Blake, Robert C., II, 186, 273 Blalock, J. Edwin, 168, 16; 178, 63 Blanchard, John S., 182, 24 Blaner, William S., 189, 193,270 Blasie, J. K., 172, 399 Bleasdale, John E., 187, 226 Blinks, John R., 172, 164 Blobel, Gianter, 194, 735 Blomhoff, Rune, 190, 58 Blostein, Rhoda, 173, 377 Blum, M., 168, 618 Blundell, Tom L., 183, 670 Boeke, Jef D., 194, 302, 774 Bolivar, Francisco, 185, 14 Boiler, Thomas, 174, 504 Bolton, Philip H., 177, 74, 282 Bonnard, Claude, 184, 629 Book, Steven A., 190, 406 Boorstein, William R., 180, 347 Borgeat, Pierre, 187, l, 98 Borgias, Brandan A., 176, 169 Bors, Wolf, 186, 343 Borst-Pauwels, George W. F. H., 174, 603 Bosch, L., 180, 289 Bossant, Marie-Jeanne, 187, 125 Bost, Kenneth L., 168, 16; 178, 63 Botstein, David, 185, 280 Boucher, Richard C., 192, 565 Bougueleret, Lydie, 183, 237

289

Bourgoin, Sylvain, 187, 98 Boursier, Patrick J., 174, 267 Bowling, D. J. F., 174, 331 Boyd, Jeff A., 186, 283 Boyd, Robert K., 193, 154 Boyer, James L., 192, 501, 517, 534 Bradley, Margaret K., 182, 112 Bradshaw, Ralph A., 185, 398 Brahm, Jesper, 173, 160, 432 Brake, Anthony J., 185, 408 Brakel, Christine L., 184, 437 Branch, Andrea D., 180, 130, 418 Brandley, Brian K., 179, 542 Br~ndli, Andr6 W., 191, 813 Brandt, W. F., 170, 431 Braquet, P., 187, 433 Brash, Alan R., 187, 187 Brass, Lawrence F., 169, 355,371 Bredberg, D. Lucille, 190, 156 Breitenbach, Michael, 194, 1l0 Breitman, Theodore R., 189, 233; 190, 118 Brendel, Volker, 183, 388 Brennan, Patrick J., 179, 215 Brewer, Gary, 181, 202 Bridges, C. D. B., 189, 60, 207, 325; 190, 141 Bridges, Sandra, 178, 356 Brill, Julie A., 194, 774 Brinckerhoff, Constance E., 190, 175 Bringmann, Peter, 181, 232 Brinkhous, K. M., 169, 149 Broach, James R., 185, 234; 194, 195 Brock, Thomas, 168, 351 Brockhausen, Inka, 179, 351 Broekman, M. Johan, 169, 415 Broker Thomas R., 180, 239 Brouwer, Adriaan, 190, 49 Brower, Mark S., 184, 437 Brown Dennis, 191, 551 Brown Elaine R., 168, 634 Brown Joseph P., 183, I II Brown L. R., 176, 199 Brown Marvin R., 168, 431 Brown. R. S., 170, 420 Bruening, George, 180, 546 Brunel, Claude, 181, 264 Brunner, Josef, 172, 628 Bryce, Graeme F., 190, 352 Buckie, J. William, 184, 304 Buell, Melissa K., 189, 363

290

CONTRIBUTOR INDEX

Buenafe, Connie A., 183, 3 Buettner, Garry R., 186, 125, 127 Bulkley, Gregory B., 186, 711 Bun, H., 189, 167 Bunschoten, E. J., 178, 375 Burbach, J. Peter H., 168, 385, 398 Burch, Paula, 186, 635 Burckhardt, Gerhard, 191, 505 Burg, M. B., 191, 226 Burgess, Richard R., 170, 612 Burk, Raymond F., 186, 759 Burke, David T., 194, 251 Burke, John M., 180, 533 Burkey, Theodore J., 186, 665 Burks, Christian, 183, 3 Burlingame, A. L., 193, 661,689 Busch, Christer, 189, 315 Bustin, Michael, 170, 214 Butlen, Daniel, 191, 303 Byers, Breck, 194, 602 Bystrykh, Leonid V., 188, 391,420,435, 445

Cabantchik, Z. I., 172, 122; 173, 250, 410 Cabelli, Diane E., 186, 116 Cadenas, Enrique, 186, 180, 287 Cadet, J., 186, 502 Cafiso, David, 171, 342; 172, 331 Caines, G. Herbert, 176, 386 Cala, Peter M., 173, 330 Cambier, John C., 168, 338 Cameron, Graham, 183, 23 Campbell, Colin R., 185, 527 Campen, Carolyn A., 168, 588 Canessa, Mitzy, 173, 176 Canfield, Louise M., 189, 418 Cano, J. P., 189, 167 Cantor, Charles R., 170, 192 Capdevila, Jorge, 187, 357, 385 Capetola, Robert J., 190, 346 Caplan, S. Roy, 171, 397 Caprioli, Richard M., 193, 214 Capron, Andre, 178, 390 Carbon, Philippe, 180, 212 Carmeliet, Peter, 168, 47 Carmichael, David F., 185, 187 Caroni, Pico, 172, 136

Carr, Steven A., 193, 501 Carrasquer, G., 171, 607 Carroll, Mairead A., 187, 365 Carter, Mark S., 168, 634 Cartwright, lain L., 170, 359 Casals-Stenzel, Jorge, 187, 455 Case, R. M., 192, 256 Casey, Joseph R., 173, 494 Catalfamo, James L., 169, 27 Catella, Francesca, 187, 42 Cathala, Guy, 181, 264 Cech, Thomas R., 181, 558 Cedergren, Robert, 183, 318, 428 Cerniglia, Carl E., 188, 148 Chabrier, P. E., 187, 433 Chader, Gerald J., 189, 213 Chalkley, Roger, 170, 612 Chambers, A., 185, 329 Chandra, Gyan, 184, 70 Chandry, P. Scott, 181, 521 Chang, Chung Nan, 185, 421 Chang, Dean, 192, 734 Chang, Hshi-Chi, 184, 497 Chang, Shih-Wen, 187, 599 Chase, Herbert S., Jr., 172, 301 Chatterjee, Delphi, 179, 215 Chauhan, Jasbir, 184, 93 Chazin, Walter J., 176, 114 Cheeseman, Kevin H., 186, 407 Chejanovsky, N., 171, 829 Chen, Christina Y., 185, 421,471 Chen, Ling-Mei, 184, 653 Chen, Ying-Nan Pan, 187, 479 Cheng, Soo-Chen, 181, 89 Chen-Kiang, Selina, 180, 69, 82 Cheret, A. M., 171, 444 Chesselet, Marie-Francoise, 168, 761 Chew, Catherine S., 191, 640 Chicz, Roman M., 182, 392 Chien, MiUie M., 168, 338 Childs, Gwen V., 184, 395 Childs, Ronald F., 189, 112 Chilton, Floyd H., 187, 157 Chinander, Laurie L., 189, 363 Chirgwin, John M., 180, 3 Chisholm, Vanessa, 185, 421,471 Chistoserdov, Andrey Y., 188, 247, 335 Choi, Yang Do, 1811, 410; 181, 317 Chow, Louise T., 180, 239 Christensen, Halvor N., 173, 576; 174, 154

CONTRIBUTOR INDEX Christodoulou, C., 168, 753 Chronwall, Bibie M., 168, 778 Church, Daniel F., 186, 665 Ciabattoni, Giovanni, 187, 34 Cigan, A. Mark, 185, 366 Cinkosky, Michael J., 183, 3 Cioffi, Moira, 192, 608 Cirillo, Vincent P., 174, 617 Citovsky, V., 171, 829 Civalier, Chris, 182, 521 Civan, Mortimer M., 192, 683 Claret, Michel, 192, 495 Clark, Michael W., 194, 608,717 Clarke, Berwyn E., 178, 659 Clarke, Steven, 172, 696 Clausen, Chris, 171, 628 Claverie, Jean-Michel, 183, 237 Clay, Keith L., 187, 134; 193, 338 Clevenger, William, 185, 308 Clifford, Andrew J., 189, 94 Climent, Isabel, 186, 464 Cohen, Irun R., 178, 300 Cohen, Jack S., 177, 435 Cohen, Leonard H., 170, 532 Cohen, Philip, 193, 480 Cohen, S., 173, 777 Cohen, Sheila M., 177, 417 Colt, Doris, 185, 341 Cole, R. David, 170, 524 Colin, Pierre, 170, 166 Collen, Desire, 169, 296 Collins, John F., 183, 474 Colman, Robert W., 169, 276 Colombatti, Marco, 178, 404 Combettes, Laurent, 192, 495 Conn, P. Michael, 168, 287 Connelly, Carla, 194, 57 Conrad, Mary K., 184, 641 Conrad-Kessel, Wendy, 187, 175 Contopoulou, C. Rebecca, 194, 827 Conway, Laura, 180, 369 Cook, Geoffrey M. W., 184, 304 Cooley, Lynn, 181, 451 Coon, Minor J., 186, 273,278 Cooper, Dale A., 189, 490 Cooper, Stuart L., 169, 91 Copello, J., 192, 650 Corey, E. J., 187, 1 Cornette, James L., 178, 611 Corrigan, Anne Z., 168, 588

291

Costanzo, Maria C., 194, 149 Costello, Catherine E., 193, 738, 875 Cotten, Matt, 170, 612 Cotter, Robert J., 193, 3 Cotton, C., 192, 650 Coulson, Andrew F. W., 183, 474 Couraud, Jean-Yves, 178, 275 Cousens, D., 185, 329 Cox, Jeffrey W., 187, 62 Crabb, David W., 168, 690 Cragoe, Edward J., Jr., 191, 739 Craig, Elizabeth A., 180, 347; 194, 710 Crain, Pamela F., 193, 782, 857 Cramer, W. A., 172, 439 Crawford, Dwight N., 192, 599 Cremins, Jean D., 168, 634 Crivellari, M. T., 187, 621 Croxton, Thomas L., 168, 144 Cull, Millard, 182, 147 Culler, Michael D., 168, 498 Cumella, Karen E., 183, 3 Cundy, Kenneth C,, 186, 521 Cuppoletti, John, 173, 392 Curci, S., 192, 82 Curnutte, John T., 186, 567 Cusick, Michael E., 170, 290 Czelusniak, John, 183, 601

D

Daban, Joan-Ramon, 170, 192 Dabrowski, Janusz, 179, 122 Daemen, Frans J. M., 189, 402 Dahinden, Clemens A., 187, 567 Dahl, Kristine D., 168, 414 Dahlquist, Frederick W., 177, 44 Dakour, J., 179, 55 Dakshinamurti, Krishnamurti, 184, 93, Ill Dalton, Howard, 188, 181 Dalvit, Claudio, 176, 114 Daly, Ann K., 189, 239, 307 Daly, Peter F., 177, 435 Dangelmaier, Carol A., 169, 195,205,336 Dantzler, William H., 191, 167 Dargemont, Catherine, 192, 495 Darrow, Andrew L., 190, 110 Das, Asis, 182, 93 Das, Goutam, 194, 362

292

CONTRIBUTOR INDEX

Davidow, Lance S., 194, 491 Davidson, Victor L., 188, 241 Davies, Christopher, 181, 583 Davies, Kelvin J. A., 186, 485 Davis, Ronald W., 194, 230 Davison, Dan B., 183, 3 Dawson, David C., 192, 734 Dawson, K., 192, 650 Dawson, M. I., 189, 15 Day, Darren J., 188, 210, 298 Dean, Ann, 170, 26 de A. Rodrigues, J., 170, 431 DeBell, Marc C., 172, 301 de Boer, A. H., 174, 277 de Boer, Herman A., 185, 103 Decarroz, C., 186, 502 De Grip, Willem J., 189, 402; 190, 3 de Groot, Cornelis, 184, 314 de Groot, Herbert, 186, 443 de Koning, Wim, 188, 435,445 Delautier, Dani~le, 187, 125 De Leenheer, Andre P., 189, 50, 104 DeLisi, Charles, 178, 61 l Dell, Anne, 193, 647 Dell'Acqua, Mark, 177, 74, 282 Dell'Arciprete, Lorena, 178, 404 De Luca, Luigi M., 190, 81, 91 de Macario, Everly Conway, 184, 501 Dencher, Norbert A., 171, 265 Denef, Carl, 168, 47 Denton, Richard M., 174, 95 DePamphilis, Melvin L., 170, 290 de Pauw, Edwin, 193, 201 Desharnals, Robert A., 168, 822 Deuel, Thomas F., 169, 233 Deuticke, Bernhard, 173, 300 Deutscher, Murray P., 181, 421,434; 182, 19, 83, 779 Dev6s, Rosa, 171, 113 Devor, Daniel C., 192, 309 DeWitt, David L., 187, 469 De Young, Mary Beth, 173, 662 Dharmsathaphorn, Kiertisin, 192, 354 Diaz, Sandra, 179, 409, 416 Diedrich, Donald F., 191, 755 Dietl, P., 192, 710 Dignam, John David, 182, 194 Dijkhuizen, L., 188, 223, 391,405 DiMaio, Daniel, 185, 599 Dishman, Elizabeth, 187, 385

• DiSpirito, Alan A., 188, 289 Distler, Jack J., 179, 304 Dixon, Jack E., 168, 72, 690 Dixon, S. J., 173, 777 Dizdaroglu, Miral, 186, 530; 193, 842 Dodds, W. Jean, 169, 27 Dolnikowski, Gregory G., 193, 37 Domdey, Horst, 194, 398 Domingo, M. T., 187, 433 Domon, Bruno, 193, 607 Doms, Robert W., 191, 841 Donahue, Thomas F., 185, 366 Doolittle, Russell F., 183, 99, 375, 659 Doran, Thomas I., 190, 334 Dorner, Andrew J., 185, 577 Dowbenko, Donald J., 185, 421 Draper, H. H., 186, 421 Dratz, Edward A., 186, 388, 399 Dresser, Michael, 194, 110 Dretzen, Guy, 170, 317 Dreyfuss, Gideon, 180, 410; 181, 317, 326 Drubin, David G., 194, 565 Drugge, Elizabeth D., 187, 365 Dubendorff, John W., 185, 60 Duffey, Michael E., 192, 309 Duhamel, Raymond C., 184, 201 Duine, J. A., 188, 33,202, 235, 260, 303, 327 Dulic, Vjekoslav, 194, 697 Dunbar, Bonnie S., 182, 441,663,670, 679 Dunn, Catherine E., 187, 365 Dunn, John J., 185, 60 Dunn, Michael J., 187, 544 Diarr, Mathias, 174, 504 Dutton, Anne H., 184, 343

Eastmond, David A., 186, 579 Eaton, Hugh L., 179, 242 Ebralidse, K. K., 170, 386 Eddy, A. Alan, 172, 95; 173, 771; 174, 623 Edmonds, Charles G., 193, 412 Edmonds, Mary, 180, 177; 181, 161 Edwards, Aled M., 194, 545 Egerton, Mark, 194, 697 Egge, Heinz, 193, 713 Ehlert, R., 190, 291 Ehrenman, Karen, 181, 521

CONTRIBUTOR INDEX Eichele, Gregor, 190, 201 Eidelman, Ofer, 172, 122 Eigen, Manfred, 183, 505 Eisenberg, David, 183, 146 Eisenberg, Shlomo, 182, 521 Elbein, Alan D., 179, 452 Elder, John H., 179, 505 Eldor, Amiram, 169, 76 Elgin, Sarah C. R., 170, 359 Elguindi, Ibrahim, 194, 697 Elias, Dana, 178, 300 Eller, Thomas D., 187, 62 Elliott, James I., 193, 389 Ellory, J. Clive, 173, 122 Emerit, Ingrid, 186, 555 Emerson, Charles H., 168, 365,567 Emmert, David B., 183, 3 Emr, Scott D., 185, 231 Emson, P. C., 168, 753 Eng, John, 168, 302 Engelke, David R., 181, 439; 194, 550 Englard, Sasha, 182, 285, 626 English, Leigh H., 173, 676 Ensley, Burr D., 188, 46 Entsch Barrie, 188, 138 Epstein, Franklin H., 192, 754 Erdmann, Erland, 173, 634 Eriksson, Ulf, 189, 315 Ernster, Lars, 186, 180, 287 Esposito, Rochelle Easton, 194, 110 Esterbauer, Hermann, 186, 407 Etcheverry, Tina, 185, 319 Etzler, Marilynn E., 179, 341 Evans, Claire F., 181, 439 Evans, Evan A., 173, 3 Evans, Ronald M., 189, 223 Evans, Syd, 193, 61 Exton, J. H., 173, 534

F

Faherty, Denise A., 190, 252 Falck, J. R., 187, 357, 385 Fanger, Bradford O., 172, 687 Farid, Nadir R., 178, 191 Farmer, B. T., II, 176, 199 Farr, Spencer B., 186, 646 Farrell, Roberta L., 188, 159 Fasold, H., 172, 584

293

Faulkner, Mary Jo, 183, 3 Feinmark, Steven J., 187, 559 Feng, Da-Fei, 183, 375,659 Fenn, Barbara J., 184, 584 Ferretti, James A., 176, 3 Ferrone, S., 178, 74 Fex, Gfran, 189, 394 Fickett, James W., 183, 3 Field, Michael, 191, 781; 192, 746 Filipowicz, Witold, 181, 148,499 Fink, Gerald R., 194, 774 Finkelstein, Eve, 184, 429 Finn, Frances M., 184, 244 Finne, Jukka, 179, 104, 270 Finnerty, W. R., 188, 10, 14, 18 Firestone, Gary L., 182, 688 Fischer, Peter W. F., 186, 232 Fischer, William M., 183, 3 Fishel, Richard, 184, 328 Fisher, Laurel A., 168, 431 Fiskum, Gary, 168, 481 Fitch, Walter M., 183, 615 FitzGerald, Garret A., 187, 42 Fitzpatrick, Frank A., 187, 553 Flanagan, Peter M., 194, 545 Fleischer, Becca, 174, 173 Fleischer, Sidney, 171, 3; 172, 410 Flemstr6m, Gunnar, 192, 139 Fotco, G. C., 187, 621 Folley, Linda S., 194, 149 Fong, Shao-Ling, 189, 207, 325; 190, 141 Forssell, Henrik, 192, 139 Forster, Anthony C., 181, 583; 184, 588 Forte, J. G., 192, 151 Foskett, J. Kevin, 171, 792 Fox, Brian G., 188, t91 Fox, Glyn R., 184, 442 Fox, Thomas D., 194, 149 Francesconi, Stephen C., 182, 521 Francis, Michael J., 178, 659 Frank, J., 188, 202, 235, 260, 303 Frankel, E. N., 186, 380 Franzusoff, Alex, 194, 662 Freedman, J. C., 172, 102; 173, 94 Frei, Balz, 186, 371 French, Edward D., 168, 103 Fridovich, Irwin, 186, 242 Friedberg, Ilan, 171, 857 Frist, William H., 186, 742 Fritz, Thomas A., 189, 418

294

CONTRIBUTOR INDEX

Frobert, Yveline, 178, 275 Froehner, Stanley C., 184, 433 Fr6hlich, Otto, 173, 54 Frojmovic, Mony M., 169, 134 Froland, Wayne A., 188, 191 Fromherz, Peter, 171, 376 Fromm, Michael, 192, 459 Fr6mter, E., 171, 642; 191, 4; 192, 82, 697 Fry, Glenna L., 169, 71 Fryktund, Jan, 191, 721 Fu, Ji Yi, 187, 486 Fuchs, Elaine V., 190, 18 Fuchs, J0rgen, 186, 197, 670, 723 Fuchs, Sara, 178, 433 Fudem-Goldin, Barbara, 184, 167 Fuhrmann, G0nter Fred, 173, 263 Fujii, Shigeru, 179, 397 Fuks, Zvi, 169, 76 Fukuda, Minoru, 179, 17 Funa, Keiko, 189, 315 Funk, Colin, 187, 486, 491 Furbacher, P. N., 172, 439 Furie, Barbara C., 169, 31 l Furie, Bruce, 169, 311 Furneaux, Henry M., 181, 480 Furr, Harold C., 189, 85, 94 Ffirstenberger, U., 172, 528 Furuichi, Yasuhiro, 180, 164

G Gait, M. J., 168, 753 Gajewski, Ewa, 186, 530 Gallacher, D. V, 192, 300 Gandini-Attardi, D., 181, 510 Garcia, Pablo D., 194, 675 Garcia-Perez, Arlyn, 171, 581 Garcia-Sancho, Javier, 173, 100, 368 Gardner, Jerry D., 171, 590; 191, 609; 192, 247, 271 Garfin, David E., 182, 425,459 Garfinkel, David J., 194, 342 Garland, Donita, 186, 464 Garrard, William T., 170, 116 Garry, Kimberly, 184, 437 Garty, Haim, 172, 155; 192, 683 Gasser, Kenneth, 174, 162 Gassner, B., 192, 710 Gautheret, Daniel, 183, 318

Gear, Adrian L., 169, 134 Geary, Philip J., 188, 52, 134 Gebeyehu, Gulilat, 184, 561 Gegenheimer, Peter, 182, 174 Geissler, Johanna F., 188, 154 Gelernt, Mark D., 172, 301 Gendimenico, Gerard J., 190, 346 Georgalis, Yannis, 172, 364 George, David G., 183, 31,333 George, Margaret A., 190, 42 Georgiev, G. P., 170, 408 Georgopoulos, Denise E., 194, 536 Gerard, Craig, 182, 529 Gerber, John G., 187, 505 Gerber, L. E., 189, 433 Gerhardt, Richard, 174, 518 Gerig, J. T., 177, 3 Gerlt, John A., 177, 74, 282 Getting, Sandra L., 194, 57 Gershengorn, Marvin C., 168, 218 Gesmar, Henrik, 176, 311 Geuze, Hans J., 169, 301 Giangrande, Angela, 170, 317 Giannattasio, Bartolomeo, 173, 662 Gibson, Bradford W., 193, 480 Gibson, Jane, 188, 154 Gigu6re, Vincent, 189, 223 Gilbert, M., 178, 144 Gillece-Castro, Beth L., 193, 689 Gilna, Paul, 183, 3 Gimeno, Carlos J., 186, 521 Giri, Lallan, 182, 380 Giudice, George J., 190, 18 Glasel, Jay A., 178, 222 Glazer, Alexander N., 184, 188; 186, 161, 301 Gliemann, JCrgen, 173, 616 Gmaj, Piotr, 172, 346 Gnann, John W., Jr., 178, 693 Goeddel, David V., 185, 3 Goedert, Michel, 168, 462 Goelz, S. E., 181, 97 Goetsch, Loretta, 194, 602 Goetz-Smith, J. D., 173, 777 Gojobori, Takashi, 183, 531 Gold, Larry, 185, l l , 89 Goldin, Stanley M., 172, 34 Goldman, June, 179, 299 Goldstein, Irwin J., 179, 327 Gollnick, H., 190, 291

CONTRIBUTOR INDEX Golovleva, L. A., 188, 115 Gonzalez, Hector A., 168, 371 Good, Mark, 183, 3 Goodall, Gregory J., 181, 148 Goodman, DeWitt S., 189, 193, 286 Goodman, Morris, 183, 601 Goodwin, Patricia M., 188, 361 Gorbunoff, Marina J., 182, 329 Gordon, Jeffrey I., 189, 506 Gordon, L. G. M., 171, 642; 192, 697 Gorenstein, David G., 177, 295 Gorman, Robert R., 187, 535 Gottesfeld, Joel M., 170, 347 Gottesman, Susan, 185, 119 Gouy, Manolo, 183, 645 Govorukhina, N. I., 188, 391 Grabo, M., 172, 528 Grabowski, Paula J., 184, 319 Gradmann, D., 174, 490 Graf, J., 192, 501 Gralla, Jay D., 185, 37 Gramsch, Christian, 178, 243 Granger, D. Neil, 186, 729 Grant, Francis J., 185, 351 Grassi, Jacques, 178, 275; 187, 24 Graupner, Gerhart, 189, 256 Gray, Gary R., 193, 573 Gray, John G., 184, 70 Green, Joanne Balmer, 190, 304 Green, Michael H., 190, 304 Green, Michael R., 181, 20, 180 Green, N. Michael, 184, 51 Green, Nordica, 171, 736 Green, Phillip R., 181, 471 Greene, Alice A., 174, 154 Greene, Mark I., 178, 321 Greer, Chris L., 181, 463 Greger, Rainer, 191, 289,793 Gregolin, Carlo, 186, 448 Grell, Ernst, 172, 364 Greyling, H. J., 170, 431 Gribskov, Michael, 183, 146 Griffin, Gail L., 169, 233 Grinstein, S., 173, 777 Grippo, Joseph F., 189, 248; 190, 148 Grisham, Matthew B., 186, 729 Griswold, Michael D., 190, 71 Groen, B. W., 188, 33, 260 Groman, Ernest V., 184, 208 Gross, Michael L., 193, 131,237

295

Grove, Gary L., 190, 360 Grove, Mary Jo, 19tl, 360 Grubbs, Robert D., 173, 546 Grygorczyk, R., 173, 112, 453 Grzych, Jean-Marie, 178, 390 Gu, Jianguo, 179, 397 Guarente, Leonard, 194, 182, 373 Gubler, Mary Lou, 189, 525 Gudas, Lorraine J., 190, 131 Guder, Walter G., 191, 325 Guerrier-Takada, Cecilia, 181, 569 Guggino, William B., 191, 371 Gunn, Robert B., 171, 164; 173, 54 Gutteridge, John M. C., 186, 1 Guy, J., 178, 308 Guyer, Cheryl A., 172, 687

H

Haarer, Brian K., 194, 565 Haas, Mark, 173, 280 Haber, Edgar, 178, 515 Hacking, A. J., 188, 379 Hadley, M., 186, 421 Haeggstr6m, Jesper Z., 187, 324 Hagenbuch, Bruno, 172, 346 Hagg, K., 192, 15 Hahn, Chang S., 180, 121 Haigler, Billy E., 188, 46 Haimovich, J., 178, 369 Hakomori, Sen-itiroh, 179, 242, 521 Hatban, Philippe A., 192, 188, 223 Hall, Elizabeth, 187, 245 Hall, Robert D., 186, 133 Halliwell, Barry, 186, 1 Halpern, Howard J., 186, 611 Haltiwanger, Robert S., 179, 82 Halushka, Perry V., 187, 397 Hamberg, Mats, 187, 1 Hamilton, Robert, 185, 421 Hamm, J6rg, 181, 273 Hammarstr6m, Sven, 187, 306 Hammel, Kenneth E., 188, 159 Hanahan, Donald J., 187, 152 Hanai, Nobou, 179, 521,242 Handler, Joseph S., 171, 736 Haner, Ronald L., 176, 418 Hanke-Baier, P., 173, 453 Hanna, Michelle M., 180, 383

296

CONTRIBUTOR INDEX

Hannig, Kurt, 171, 513 Hanrahan, John W., 192, 632 Hansen, Carl A., 191, 691 Hansen, William, 194, 675 Hansson, Gunnar C., 193, 733 Harder, Wim, 188, 420, 435,445 Hardy, Mark R., 179, 65, 76 Harpel, Mark R., 188, 101, 107 Harpel, Peter C., 169, 268 Harpold, Lynn, 177, 74, 282 Harrington, Michael G., 182, 488 Harris, Nomi L., 183, 252 Harrison, Alex G., 193, 3 Harrison, Earl H., 189, 459 Hart, Gerald W., 179, 82 Hartley, Donna L., 182, 264 Hartman, Philip E., 186, 310 Harvey, Catherine M., 173, 122 Harvey, William J., 173, 377 Harvey, William R., 192, 599, 608 Harwood, Caroline S., 188, 154 Hascall, Vincent C., 179, 435 Hasegawa, Masami, 183, 550 Hashimoto, Nobuo, 184, 497 Haslam, Richard J., 169, 455 Hassan, Ahmed H. S., 178, 243 Hawiger, Jacek, 169, 1l, 191 Hawkins, Dan J., 187, 187 Hayashi, Yutaro, 172, 514 Hayden, Jamie E.-D., 183, 3 Hayes, Edward C., 187, 116 Hayes, Jeffrey J., 186, 545 Hayes, Roger N., 193, 237 Haywood, Geoffrey W., 188, 427 Hazum, Eli, 168, 494, 729; 184, 285 Heard, Gregory S., 184, 103 Heaton, W. Andrew, 169, 172 Hedin, Lars O., 194, 149 Hedrich, Rainer, 174, 312 Heffron, Fred, 194, 329 Hegel, Ulrich, 192, 459 Heidrich, Hans-G, 171, 513 Hein, Jotun, 183, 626 Heinemann, Jack A., 194, 187 Heitkamp, Michael A., 188, 148 Hekman, Carla M., 168, 422 Heldt, Hans W., 174, 518 Heller, Werner, 186, 343 Hellerqvist, Carl G., 193, 554 Helman, Joseph, 192, 26

Hempling, Harold G., 172, 262 Henderson, James S., 186, 686 Hendriks, Henk F. J., 190, 49 Henikoff, Steven, 183, 111 Henner, Dennis J., 185, 54, 199, 223 Henson, Peter M., 187, 130 Heppel, Leon A., 171, 857 Herbette, L. G., 172, 399 Herman, Timothy M., 170, 41; 184, 584 Hermann, Klaus, 168, 545 Hermann, Thomas, 189, 256 Herrick, David, 194, 415 Hersey, Stephen, 192, 124 Herskowitz, Ira, 194, 132 Herz, Albert, 178, 243 Heuer, Hubert O., 187, 455 Heyens, Anthon du P., 169, 172 Heyn, Maarten P., 172, 462, 575 Hibler, David W., 177, 74, 282 Hicks, James, 194, 21 Hieter, Philip, 194, 57, 749 Hiewiarowski, Stefan, 169, 224 Higa, Herman H., 179, 409, 416 Higgins, David R., 194, 319 Hill, Donald L., 189, 43 Hill, John E., 194, 477 Hill, Kristina E., 186, 759 Hillenkamp, Franz, 193, 280 Hiller, Yaffa, 184, 68 Hir, Michel Le, 191, 265 Hiramatsu, Midori, 190, 273 Hitzeman, Ronald A., 185, 421,471 Hjelmeland, Leonard M., 182, 253,277 Hmiel, S. Paul, 173, 546 Hoak, John C., 169, 71 Hobbs, P. D., 189, 15 Hoch, Jeffrey C., 176, 216 Hoch, Sallie O., 181, 257 Hochstein, Paul, 186, 287 Hoekstra, Merl F., 194, 329 Hof, D., 173, 112 H6fer, Milan, 174, 629 Hoffmann, Karen S., 173, 330 Hofmann, Kathryn J., 185, 297 Hofmann, Klaus, 184, 244 Hofmann, Klaus H., 188, 451 Hokin, Lowell E., 191, 676 Holland, I. Barry, 182, 132 Holmsen, Holm, 169, 195,205, 336

CONTRIBUTOR INDEX Holt, Gordon D., 179, 82 Holt, John C., 169, 224 Homo, Jean-Claude, 170, 166 Honda, Zen-ichiro, 187, 296 Hopfer, Ulrich, 172, 313; 174, 162; 192, 389 Hopkins, Pamela, 174, 623 Hopp, Thomas P., 178, 571 Hopper, Anita K., 181, 400 Hopper, James E., 185, 297 Horan, Paul Karl, 171, 549 Hore, P. J., 176, 64 Horecker, B. L., 182, 788 Horn, Valerie J., 192, 26 Horne, Deborah A., 183, 3 Horne, William C., 173, 380 Horowitz, Paul, 184, 234 Horster, Michael F., 191, 409 Horwitz, Arnold H., 178, 476 Horwitz, Bruce H., 185, 599 H6rz, Wolfram, 170, 630 Hosford, D. J., 187, 433 Hostomsky, Z., 181, 97 Houghton, F. Kay, 183, 3 Howard, W. Brian, 190, 418, 427, 433 Hsu, Su-Ming, 184, 357 Hsueh, Aaron J. W., 168, 414, 422 Huang, Freesia L., 190, 91 Huang, Jung San, 169, 233 Huang, Sue-Ying, 170, 116 Hubbard, Ann L., 191, 825 Hughes, Helen, 186, 681 Hui, Anna S., 185, 103 Huibregtse, Jon M., 194, 550 Hull, Eric, 179, 351 Humm, Jane, 168, 351,358 Humphries, Donald E., 179, 422, 428 Hunt, Lois T., 183, 31, 333 Hunter, Shirley Wu, 179, 215 Hurley, James, 190, 326 Hurley, Walter L., 184, 429 Hurwitz, E., 178, 369 Hurwitz, Jerard, 181, 480 Husain, Mazhar, 188, 309 Huskins, Kenneth R., 184, 556 Hussaini, I. M., 187, 446 Hutchins, Cheryl J., 181, 583 Hymer, W. C., 168, 327 Hymes, Jeanne, 184, 103

297

Ichikawa, Iekuni, 191, 107 Igarashi, Yasuyuki, 179, 521 Ii, Masayuki, 179, 322 lkemura, Toshimichi, 180, 14 Inagami, Tadashi, 191, 152 Ingham, Kenneth C., 182, 301 Irvine, Robin F., 191, 707 Issels, Rolf D., 186, 696 Ito, Makoto, 179,488 Itoh, Kiyoshi, 169, 430 lzumi, Takashi, 187, 286, 296

Jacobson, Allan, 194, 415 Jacobson, Kenneth A., 184, 668 Jacquemin, Marc G., 178, 150 Jaeger, John A., 183, 281 Jaeschke, Hartmut, 186, 681,752 Jaggi, P., 172, 528 James, Bryan D., 180, 227 James, Thomas L., 176, 169 Jardetzky, Oleg, 177, 218 Jardine, lan, 193, 441 Jazwinski, S. Michal, 182, 154 Jefferies, Leigh C., 179, 299 Jencks, William P., 171, 145 Jennings, Keith R., 193, 37 Jensen, Bruce D., 171, 549 Jensen, Robert E., 194, 132 Jensen, Robert T., 171, 590; 192, 247. 271 Jentsch, Thomas J., 192, 571 Jessee, Bret, 170, 603 Jetten, Anton M., 189, 248; 190, 42 Jia, Xiao-Chi, 168, 414 Joannou, Chris L., 188, 52 Johannesson, Gunvor, 189, 394 Johnson, E. R., 173, 771 Johnson, Mark S., 183, 670 Johnson, Peter C., 169, 386 Johnson, Robert M., 173, 35 Johnstone, Rose M., 171, 593 Joist, J. Heinrich, 169, 172 Jolley, Diane, 168, 588 Jollie, David R., 188, 191, 331 Jones, A. Daniel, 189, 94 Jones, David A., 187, 553

298

CONTRIBUTOR INDEX

Jones, Elizabeth W., 185, 372; 194, 428 Jones, Robert, 183, 221 Jones, T. Alwyn, 189, 281 Jongejan, J. A., 188, 260 J6rnvall, Hans, 187, 486 Joseph, Shirley A., 178, 212 Jourdian, George W., 179, 304 Jovell, Robert J., 184, 501 Jovin, Thomas M., 172, 471 Judd, Ralph C., 182, 613 Julia, Pere, 189, 436 Jung, Chan Y., 173, 392 Jungalwala, F. B., 172, 538

Kagan, Valerian E., 186, 355 Kageshita, T., 178, 74 Kahn, Patricia, 183, 23 Kaissling, Brigitte, 191, 265 Kajikawa, Norio, 169, 430 Kaku, Hanae, 179, 327 Kalyanaraman, B., 186, 333; 188, 159 Kam, Laurance, 186, 545 Kambayashi, Jun-Ichi, 169, 442 Kans, Jonathan A., 194, 827 Kaplan, Lawrence A., 189, 155 Kaplan, Ronald S., 172, 393; 173, 732 Kaplowitz, Nell, 192, 485 Kaptein, R., 177, 204 Karara, Armando, 187, 385 Karas, Michael, 193, 280 Karl, Alice F., 190, 71 Karlin, Samuel, 183, 388 Karlish, S. J. D., 172, 155; 191, 469 Karlsson, Hasse, 193, 733 Karisson, Karl-Anders, 193, 623 Karpov, V. L., 170, 386 Kassir, Yona, 194, 94 Kastin, Abba J., 168, 652 Kataeva, I. A., 188, 115 Kato, Michimasa, 189, 330 Kato, Nobuo, 188, 397,455,459 Kato, Shigemi, 190, 81 Katopodis, Andreas G., 188, 3 Kaufman, Randal J., 185, 487, 537, 577 Kaushal, Gut P., 179, 452 Kaveri, Srinivas, 178, 3

Kawasaki, Hiroshi, 187, 486 Kawasaki, Nobuko, 179, 310 Kawasaki, Toshisuke, 179, 310, 322 Kealey, Terence, 190, 338 Kedzierski, Wojciech, 168, 371 Kehoe, Suzanne M., 183, 601 Kehrer, James P., 186, 621 Keil, Borivoj, 183, 50 Kelkar, Praful M., 183, 3 Kelleher, Raymond J., III, 194, 545 Keller, Walter, 181, 3 Kelley, Tom A., 183, 3 Kelly, Michael, 183, 3 Kelsay, Kimberly, 185, 351 Kempner, E. S., 172, 410 Kendall, Marcia E., 168, 327 Kendall, Richard L., 185, 398 Kendrick, Keith M., 168, 182 Keniry, Max A., 176, 376 Kennedy, Robert M., 182, 339 Kennedy, Ronald C., 178, 36 Kenny, Brendan, 182, 132 Keown, Wayne A., 185, 527 Kersten, Philip J., 188, 159 Kieber-Emmons, Thomas, 178, 3 Kikkawa, Ushio, 168, 347; 169, 430 Kilberg, Michael S., 173, 564 Kim, I1 Han, 186, 478 Kim, Kanghwa, 186, 478 Kimmich, George A., 192, 324 Kimura, Akiro, 169, 233 Kimura, Hitomi, 182, 441 Kimura, Motoo, 183, 531 Kincaid, Randall L., 184, 451 King, Melinda A., 183, 3 King, Steven C., 172, 34 Kingsman, A. J., 185, 329 Kingsman, S. Me, 185, 329 Kinlough-Rathbone, Raelene L., 169, 3 Kinne, Rolf K. H., 172, 3; 191, 380, 450 Kinne-Saffran, Evamaria, 172, 3; 191, 450 Kinoshita, Noriaki, 179, 570 Kiriukhin, Michael Y., 188, 247, 335 Kirk, T. Kent, 188, 159 Kish, Phillip E., 174, 9 Kishino, Hirohisa, 183, 550 Kistler, Andreas, 190, 418,427, 433 Kizer, John S., 168, 351,358 Klar, Amar, 194, 795 Klaus, Michael, 189, 3

CONTRIBUTOR INDEX Klein, Ulla, 192, 608 Kletsova, Ludmila V., 188, 335 Klevan, Leonard, 184, 561 Kleyman, Thomas R., 191, 739 Kligman, Lorraine H., 190, 372 Klingenberg, Martin, 171, 12 Klootwijk, Jacobus, 180, 96 Klug, A., 170,420 Knapp, Daniel R., 187, 62; 193, 314 Knapp, Gayle, 180, 110, 192 Knauf, H., 192, 15 Knauf, Philip A., 173, 432 Knepper, M. A., 191, 226 Knigge, Karl M., 178, 212 Knight, Derek E., 171, 817 Knook, Dick L., 190, 49 Kobata, Akira, 179, 46, 261,331 Kochhar, Devendra M., 190, 317 Koepsell, H., 191, 437, 583 Koerner, T. J., 194, 477 Koger, Jeanne B., 168, 358 Kohanski, Ronald A., 184, 194 Kohen, Fortune, 184, 481 K6hler, Heinz, 178, 3 Kohler-Staub, Doris, 188, 355 Kohno, Tadahiko, 185, 187 Kohr, William J., 185, 421 K6hrer, Karl, 194, 398 Koide, Yoshio, 178, 422 Kolb, H.-A., 171, 274 Kolb-Bachofen, Victoria, 179, 111 Kolodziej, Peter A., 194, 508 Komor, Ewald, 174, 288, 552 Konarska, Maria M., 180, 442 Kondo, Takahito, 171, 217 Kopp, David W., 180, 177 Koppenol, W, H., 186, 148 Kornberg, Arthur, 182, 1,783 Kornberg, Roger D., 170, 3; 181, 332; 194, 545 Kosen, Phyllis A., 177, 86 Koshland, Douglas, 194, 749 Kosin, Sabine, 178, 243 Kottra, G., 171, 642; 191, 4; 192, 697 Kotyk, Arno~t, 174, 567, 592 Koval, Michael, 194, 749 Kraehenbuhl, Jean-Pierre, 184, 629 Kr~imer, Angela, 181, 3,215,284 Kramer, Reinhard, 171, 387 Krause, James E., 168, 634

Krause, Rudolph G., II, 168, 808 Krchfifi.k, Viktor, 178, 586 Kreisberg, J. I., 191, 141 Krema, Cinder, 188, 373 Kriegler, Michael, 185, 512 Kris, Richard M., 168, 481 Krol, Alain, 180, 212 Krolick, Keith A., 178, 448 Kriiger, Ulrich, 184, 518 Krupka, R. M., 171, 113 Kucherlapati, Raju S., 185, 527 Kuchino, Y., 180, 154 Kulmacz, Richard J., 186, 431 Kuntz, I. D., 177, 159 Kuranda, M. J., 194, 682 Kushner, Leslie, 169, 104 Kussie, Paul H., 178, 49.91

L'Abb6, Mary R., 186, 232 LaGamma, Edmund F., 168, 681 Laine, Roger A., 179, 157; 193, 539 Lakshman, M. Raj, 189, 425 Lambert, Willy E., 189, 104 Lambie, Eric J., 194, 146 Lanave, Cecilia, 183, 570 Landau, Gad M., 183, 487 Lander, D. J., 191, 707 Landers, Gary M., 189, 70 Landry, Donald W., 191, 572 Lands, William E. M., 186, 431 Lane, M. Daniel, 184, 194 Langan, Bernard J., 183, 3 LaPointe, Jan W., 170, 3 Laposata, Michael, 187, 237 Large, Peter J., 188, 427 Laris, Philip C., 171, 593 LaRochelle, William J., 184, 433 Laskey, Ronald A., 170, 575 Latham, John A., 181, 558 L~iuchli, Andr6, 174, 267 Laue, Thomas M., 182, 555,566 Lauer, Jeffrey T., 183, 3 L~iuger, P., 171, 274 Lavery, Daniel J., 180, 69, 82 Lawrence, Charles B., 183, 133 Lawrence, Christopher W., 194, 273 Lazarus, Lawrence H., 168, 444

299

300

CONTRIBUTOR INDEX

Leader, John P., 171, 744 Le Breton, Guy C., 187, 245, 406 Led, Jens J., 176, 311 Lee, Kevin A. W., 181, 20 Lee, Reiko T., 179, 253, 257 Lee, Terry D., 193, 361 Lee, Yuan C., 179, 65, 253, 257 Lee-Stadelmann, Ok Young, 174, 225, 246 Left, Jonathan A., 186, 651 Legrain, P., 181, 122 Le Grice, Stuart F. J., 185, 201 Lehmann, Juergen M., 189, 256 Leibowitz, Michael J., 194, 536 Leisinger, Thomas, 188, 355 Lellouche, J. P., 187, 70, 82 LeMaster, David M., 177, 23 Lennox, Ronald W., 170, 532 Lenz, Anke-G., 186, 464 Leo, M. A., 189, 483,520 Leong, Denis A., 168, 263 Leong, Merlin M. L., 184, 442 Leslie, Christina C., 187, 216 LeStourgeon, Wallace M., 181, 293 Leto, Thomas L., 173, 380 Leung, Lawrence L. K., 169, 268 Levenson, Corey, 184, 577 Levering, P. R., 188, 405 Levery, Steven B., 179, 242 Levi, Roberto, 187, 610 Levin, Marc S., 189, 506 Levine, Jon E., 168, 166 Levine, Richard F., 169, 76 Levine, Rodney L., 186, 464 Levinson, Arthur D., 185, 485 Levy, Daniel, 174, 25 Lew, Virgilio L., 173, 100 Lewin, Miguel J. M., 171, 444 Lewis, Michael E., 168, 761,778, 808 Lewis, Simon A., 171, 715; 192, 632 Li, Ellen, 189, 506 Li, Maomi, 179, 304 Li, Su-Chen, 179, 479 Li, Wen-Hsiung, 183, 645 Li, Yu-Teh, 179, 479 Lidstrom, Mary E., 188, 365, 373 Lieber, C, S., 189, 483,520 Lieber, Michael R., 173, 356 Lieberman, Debra M., 173, 494 Liedtke, Carole M., 192, 549 Lienhard, Gustav E., 174, 39 Lilley, Ross McC., 174, 518

Lim, Chang T., 187, 406 Lin, Ren-Jang, 181, 89 Lin, Zheng-shi, 189, 325 Lind, Christina, 186, 287 Linden, Carol D., 168, 481 Lindon, Jack N., 169, 104 Lindsey, G. G., 170, 431 Linn, Stuart, 182, 9 Linthicum, D. Scott, 178, 49, 91 Lipscomb, John D., 188, 82, 89, 95, 101, 107, 191,331 Liu, Bin, 168, 385 Liu, Chung, 185, 421 Livnah, Oded, 184, 90 Livrea, Maria A., 189, 369, 503 Lo, Mathew M. S., 184, 641 London, Robert E., 176, 358 Long, Antony R., 188, 216 Longo, Dan L., 178, 356 Lopez, Natalie, 183, 3 Lopez-Fernandez, Mafia Fernanda, 169, 244 Lorch, Yahli, 170, 3 Lotan, Dafna, 190, 100 Lotan, Reuben, 190, 100, 217 Love, Ted W., 178, 515 Low, Walter C., 168, 234 Loyter, Abraham, 171, 829 Lucas, William J., 174, 443 Luchinat, Claudio, 177, 246 Ludwig, David S., 178, 163 Lue, Neal F., 194, 545 Liihrmann, Reinhard, 181, 232 Lumsden, Charles J., 191, 34 Lunazzi, Gian Carlo, 174, 50 Lund, Elsebet, 181, 36 Lundblad, A., 179, 55 Liischer, Bernhard, 181, 74 Lustig, Kevin G., 171, 857 Li~thy, Roland, 183, 146 Lutter, Leonard C., 170, 264 Lynch, Conrad, 183, 3 Lynch, Janet, 183, 3 Lyon, Robbe C., 177, 435

M

Ma, Hong, 185, 280 Maack, Thomas, 191, 340 Macario, Alberto J. L., 184, 501

CONTRIBUTOR INDEX MacCrehan, William A., 189, 172 MacDonald, Margaret R., 168, 634 MacDonald, Paul N., 189, 450 MacFarlane, Ronald D., 193, 263 MacKay, Vivian L., 185, 351 Mach, Otakar, 178, 586 Machen, Terry E., 192, 38 Macher, Bruce A., 179, 167 Macknight, Anthony D. C., 171, 744 Maclouf, Jacques, 187, 24, 82 Madara, James L., 192, 354 Madden, Edward A., 182, 203 Maddipati, K. R., 187, 268 Maddrell, S. H. P., 192, 617 Maden, Malcolm, 190, 189 Maertens, Philippe, 168, 47 Mager, W. H., 194, 453 Maguire, Michael E., 173, 546 Maillet, Sylvie, 178, 275 Maiorino, Matilde, 186, 448 Mais, Dale E., 187, 397 Major, Franqois, 183, 318 Malathi, Parameswara, 192, 438 Malinowska, Danuta H., 192, 108 Malmstr6m, K., 191, 494 Ma137, Antonfn, 178, 586 Mannervik, Bengt, 187, 306 Manzi, Adriana, 179, 409 Marchetti, Dario, 178, 91 Marchi, Janet B., 183, 3 Marcus, Aaron J., 187, 585 Margalit, Hanah, 178, 611 Markakis, Diane, 191, 371 Markley, John L., 176, 12 Marklund, Stefan L., 186, 260 Marnett, Lawrence J., 187, 479; 190, 281 Marr, Thomas G., 183, 3 Marston, Fiona A. O., 182, 264 Martin, Bernard, 189, 239 Martin, Joseph P., Jr., 186, 220, 635 Martin, Stephen A., 193, 519 Martin, Thomas F. J., 168, 225 Martinez, Frances A., 183, 3 Martinez, Hugo M., 183, 306 Martfnez de la Escalera, Gonzalo, 168, 254 Marver, Diana, 191, 520 Marzluff, William F., 181, 30 Maskos, Z., 186, 148 Mason, Jeremy R., 188, 52, 134 Mason, Ronald P., 186, 127, 318 Masters, Bettie Sue Siler, 187, 253

301

Matarese, Valerie, 189, 363 Mather, Jennie P., 185, 567 Mathews, Christopher K., 182, 539 Mathews, W. Rodney, 187, 76 Matsudaira, Paul, 182, 602 Matsui, Hideo, 172, 514 Matsumoto, Takashi, 187, 486,491 Mattaj, lain W., 181, 273 Mattoccia, E., 181, 510 Matunis, Michael J., 181, 326 Mauger, Jean-Pierre, 192, 495 Maxfield, Frederick R., 173, 745 May, Sheldon W., 188, 3 Mayo, Laura A., 186, 567 McArdle, Craig A., 168, 287 McCabe, Joseph T., 168, 822 McCann, S. M., 168, 716 McClintock, Richard, 168, 588 McCloskey, James A., 193, 329, 771, 796, 825, 869, 882 McCluer, R. H., 172, 538 McCormack, James G., 174, 95 McCray, J., 178, 676 McDonagh, Antony F., 186, 301 McFarland, Gloria D., 181, 89 McGiff, John C., 187, 365 McGivan, John D., 174, 31 McGuire, James C., 187, 226 McGuire, John L., 190, 346 McHenry, Charles S., 182, 147 McInnes, James L., 184, 588 McIntire, William S., 188, 227, 250 Mclntosh, Lawrence P., 177, 44 McIntyre, Thomas M., 187, 344, 520 McLaughlin, Alan, 171, 342 McLaughlin, Stuart, 171, 342 McLeod, Mia J., 183, 3 McManus, Thomas J., 173, 280 McMichens, R. B., 171, 286 McMullin, Thomas W., 194, 149 McMurray, Walter, 193, 389 McNamee, Mark G., 182, 499 McNeil, Michael, 179, 215 Meagher, Richard B., 188, 126, 130 Meda, Paolo, 192, 188,223 Meguro, Hiroshi, 186, 157 Mehlhorn, Rolf J., 186, 197,670 Mehta, Rajendra G., 190, 395 Meier, Kathryn E., 184, 660 Meier, Peter J., 192, 517, 534 Meiri, Hamutal, 178, 714

302

CONTRIBUTOR INDEX

Meissner, Hans Peter, 192, 235 Melhem, Randa F., 180, 177 Melton, Doug A., 180, 42 Melvin, James E., 192, 26 Menozzi, Delia, 192, 271 Merril, Carl R., 182, 477 Merrill, Walter H., 186, 742 Mertz, Lawrence M., 192, 26 Mezick, James A., 190, 346 Mezzetti, M., 187, 621 Michael, Gregory J., 178, 212 Michel, Christa, 186, 343 Miethe, Dietmar, 188, 346 Mignon, Annick, 168, 47 Miki, Ichiro, 187, 296 Milberg, Richard M., 193, 305, 870 Mildvan, Albert S., 177, 333 Miles, Lindsey A., 169, 296 Milich, David R., 178, 634 Miller, Dennis M., 186, 457 Miller, James F., Jr., 186, 431 Miller, Judith A., 189, 155 Miller, Melissa, 192, 124 Milligan, John F., 180, 51 Milstone, Leonard M., 190, 76 Milton, John G., 169, 134 Minami, Michiko, 187, 486 Minth, Carolyn D., 168, 690 Mircheff, Austin K., 172, 18; 192, 341 Mirzabekov, A. D., 170, 386 Misevic, Gradimir N., 179, 95 Mishra, Santosh K., 183, 3 Mitchell, Jerry R., 186, 681,752 Mocali, Alessandra, 186, 209 Mock, Donald M., 184, 224, 234 Moerschell, Richard P., 194, 362 Moir, Donald T., 194, 491 Moldave, Kivie, 182, 809 Moloshok, Thomas, 179, 566 Monck, Jonathan R., 191, 691 Moncrief, Nancy D., 183, 601 Mondragon, Socorro M., 183, 3 Mong, Seymour, 187, 421 Moody, Terry W., 168, 481 Moon, Richard C., 190, 395 Moore, Claire L., 181, 49 Moore, John, 183, 3 Moore, William T., 193, 214 Morel, Francois, 191, 303 Moreno, Sergio, 194, 795 Morgan, Courtney F., 176, 386

Morinelli, Thomas A., 187, 397 Moriyama, Etsuko N., 183, 531 Morris, Randal E., 184, 379 Morrison, Sherie L., 178, 459 Morrow, Jason D., 187, 51 Morrow, W. John W., 178, 3 Mortimer, Robert K., 194, 827 Mosher, Deane F., 169, 91 Moss, Bernard, 181, 170 Mozaffar, Sabiha, 188, 463 Muchmore, David C., 177, 44 Muerhoff, A. Scott, 187, 253 Muggli, Reto, 169, 37 Muirhead, E. Eric, 191, 152 Mulero, Julio J., 194, 149 Mullane, Kevin M., 187, 610 Miiller, Dieter R., 193, 607 Mtiller, Roland H., 188, 401 Miiller, Sybille, 178, 3 Muller, Sylviane, 170, 251 Miiller-Kraft, Gabriele, 188, 350 Munk, Christine A., 183, 3 Munkres, Kenneth D., 186, 249 Murai, Ichiro, 168, 234 Muramatsu, Takashi, 179, 496 Murata, Mitsuo, 183, 365 Muter, H., 172, 346; 191, 494 Murphy, Michael E., 186, 595 Murphy, Robert C., 187, 90; 193, 338 Murrell, J. C., 188, 21, 26 Musch, Mark W., 191, 781; 192, 746 Mustard, J. Fraser, 169, 3 Muto, Yasutoshi, 189, 330 Myers, Alan M., 194, 477 Mylin, Lawrence M., 185, 297

N Nachman, Ralph L., 169, 268 Nagarajan, Vasantha, 185, 214 Nagele, Arno, 186, 696 Nakano, Minoru, 186, 227, 330, 585 Nakayama, Grace R., 178, 551 Napoli, Joseph L., 189, 459, 470 Nasseri, Kevin K., 183, 3 Nan, Heinz, 190, 437 Nauss, Kathleen M., 190, 259 Nedospasov, S. A., 170, 408 Needham, Leila K., 179, 542 Neff, W. E., 186, 380

CONTRIBUTOR INDEX Negro-Vilar, AndrOs, 168, 498, 517 Negulescu, Paul A., 192, 38 Neidle, Ellen L., 188, 122 Nelis, H. J., 189, 50 Nelles, Mitchell J., 178, 171 Nelson, Debra, 183, 3 Nelson, Will, 183, 3 Nerurkar, Lata S., 184, 541 Nervi, Clara, 189, 248 Neubauer, Bernd, 170, 630 Neubort, Simon, 168, 574 Neugebauer, Judith M., 182, 239 Neumann, Jeanne E., 184, 556 Newcomer, Marcia, 189, 281 Newman, Andrew, 181, 89 Newman, Walter, 184, 275 Ngai, Ka-Leung, 188, 122, 126, 130 Nguyen, Tan, 183, 3 Niccolai, Neri, 176, 184 Nicholls, David G., 174, 3, 85 Nichols, Mark, 181, 377 Nickoloff, Jac, 194, 329 Nicolet, Charles M., 194, 710 Niewiarowski, Stefan, 169, 22 Niki, Etsuo, 186, 100, 330 Nilsson, Bj6rn, 185, 144 Nilsson, Bo, 193, 587 Ninio, Ewa, 187, 125 Nir, Izhak, 184, 370 Nishikawa, Atsushi, 179, 397 Nishimura, S., 180, 154 Nishizuka, Yasutomi, 168, 347; 169, 430 Nojiri, Hisao, 179, 521 Noll, Hans, 170, 55 Noll, Markus, 170, 55 Nordblom, Gerald D., 186, 273 Nordlinder, Hans, 189, 315 Nores, Gustavo A., 179, 242 Northington, Stephanie J., 181, 293 Novak, I., 192, 3 Novak, T. S., 172, 102; 173, 94 Nurse, Paul, 194, 795 Nussinov, Ruth, 183, 487

O

Oberleithner, H., 191, 437; 192, 710 Odeimat, Abdulrahman, 187, 98 OdenwaUer, Rebecca, 187, 479 O'Donohue, Thomas L., 168, 3

303

Ogita, Kouji, 168, 347 O'Grady, Scott M., 191, 781; 192, 746 Ohishi, Nobuya, 187, 286 Ohkuma, Shoji, 174, 131 Ohlson, S., 179, 55 Ohno, Mutsuhito, 181, 209 Ohno, Shigeo, 187, 486 Ohrui, Hiroshi, 186, 157 Okuno, Masataka, 189, 330 Oldstone, Michael B. A., 178, 693 Olins, Peter O., 185, 115 Oliver, Cynthia N., 186, 464, 575 Ollis, David, 182, 646 Olsen, Gary J., 180, 227 Olson, James Allen, 189, 425 Olson, Maynard V., 194, 251 Ong, David E., 189, 450 Ono, R., 178, 74 Ookhtens, Murad, 192, 485 Op den Kamp, Jos A. F., 173, 223 Opella, S. J., 176, 242 Oppenheimer, Norman J., 176, 78 Orfanos, C. E., 190, 291 Orlean, P., 194, 682 Orlich, Gabriele, 174, 288 Ornston, L. Nicholas, 188, 122, 126, 130 Orr, George A., 184, 167 Orrenius, Sten, 186, 627 Orville, Allen M., 188, 82, 89, 107 O'Shannessy, Daniel J., 184, 162 Osheim, Yvonne N., 180, 481 Oshiro, C. M., 177, 159 Osipchuk, Y., 192, 300 Osterhaus, A. D. M. E., 178, 375 Ostroff, Gary, 179, 275 Ostrove, Steven, 182, 357, 371 Oudet, Pierre, 170, 14, 166 Overton, J. A., 192, 617 Ozols, Juris, 182, 225, 587

Pace, Bernadette, 181, 366 Pace, Norman R., 18tl, 227; 181, 366 Pace-Asciak, C. R., 187, 1 Pacifici, Robert E., 186, 485 Packer, Lester, 186, 197, 670, 723; 190, 273 Packham, Marian A., 169, 3 Pagano, Richard E., 171, 850

304

CONTRIBUTOR INDEX

Pai, Roger, 185, 421 Paine, Philip L., 181, 36 Palmer, Robert E., 194, 749 Palovcik, Reinhard A., 168, 129 Pantazis, Panayotis, 169, 210 Paoletti, Francesco, 186, 209 Papahadjopoulos, Demetrios, 171, 193 Papermaster, David, 184, 370 Papp, Audrey C., 187, 578 Parrs, Xavier, 189, 436 Park, C. Hyung, 191, 340 Park, Jeen-Woo, 186, 521 Park, Kinam, 169, 91 Parker, Charles W., 182, 700, 721 Parker, John C., 173, 292 Parker, Katherine A., 180, 454 Parker, Roy, 180, 510; 194, 415 Passaniti, Antonino, 179, 82 Passow, H., 173, 453 Patrignani, Paola, 187, 34 Patrono, Carlo, 187, 34 Patton, George M., 187, 195 Paul, Cynthia P., 180, 418 Payne, Lendon G., 178, 746 Payne, N. Ann, 187, 505 Pearson, William R., 183, 63 Pedersen, Peter L., 172, 393; 173, 732 Pederson, David S., 170, 26 Peerce, Brian E., 192, 165 Pelkonen, Sinikka, 179, 104 Peltz, Stuart W., 194, 415 Pendleton, Robert B., 186, 431 Pennypacker, Keith R., 184, 451 Perlman, Philip S., 181, 539 Perosa, F., 178, 74 Pesole, Graziano, 183, 570 Peter-Katalinic, Jasna, 193, 713 Petersen, O. H., 171, 663; 192, 300 Peterson, A. A., 172, 439 Peterson, Glenn, 193, 389 Petit, C., 178, 144 Pfaff, Donald W., 168, 822 Pfahl, Magnus, 189, 256 Philippsen, Peter, 194, 169 Phillips, J. M., 192, 501 Phillips, M. Ian, 168, 129, 545 Phizicky, Eric M., 181, 463 Picard, Didier, 194, 389 Picard, Serge, 187, 98 Pick, Leslie, 181, 480

Pickett, Walter C., 187, 142 Piekut, Diane T., 178, 212 Pietrobon, Daniela, 171, 397 Pikielny, C., 181, 122 Pilkington, Simon J., 188, 181 Pimlott, Weston, 193, 623 Pino, Richard M., 184, 388 Pifiol-Roma, Seraffn, 180, 410; 181, 317, 326 Pisoni, Ronald L., 174, 154 Pitcock, James A., 191, 152 Plagemann, Peter G. W., 173, 714 Planta, R. J., 180, 96; 194, 453 Pleij, C. W. A., 180, 289 Plow, Edward F., 169, 296 Pliickthun, Andreas, 178, 497; 182, 132 Pohl, Jan, 192, 124 Pohl, Thomas, 182, 68 Polak, J. M., 168, 791 Pollack, Scott J., 178, 551 Polli, Joseph W., 184, 451 Pomerantz, Steven C., 193, 796 Ponec, Maria, 190, 30 Porter, John C., 168, 371 Posnett, David N., 178, 739 Poubelle, Patrice E., 187, 98 Poulter, Linda, 193, 661 Pourmotabbed, Tayebeh, 177, 74, 282 Powell, Keith D., 168, 166 Pozzan, Tullio, 172, 230 Pradelles, Philippe, 178, 275; 187, 24, 82 Pratt, Raymond D., 173, 732 Preobrazhenskaya, O. V., 170, 386 Preparata, Giuliano, 183, 570 Prescott, Stephen M., 187, 344, 520 Pretlow, Theresa P., 171, 462 Pretlow, Thomas G., 171, 462 Price, Virginia L., 185, 308 Pringle, John R., 194, 565, 729, 732 Pryor, William A., 186, 665 Puglisi, Joseph D., 180, 304 POschel, B., 191, 437 Q Quamme, Gary A., 191, 72 Quarles, Richard H., 179, 291 Quayle, J. R., 188, 379 Quertermous, Thomas, 178, 515

CONTRIBUTOR INDEX

Rabon, E., 191, 469 R~.dmark, Olof, 187, 296,486 Raizada, Mohan K., 168, 545 Ramain, Philippe, 170, 317 Ramesha, Chakkodabylu S., 187, 142 Ramwell, Peter W., 187, 1 Rance, Mark, 176, 114 Randhawa, Zafar I., 184, 275 Rando, Robert R., 189, 411,494 Rangwala, Shaukat H., 185, 115 Rao, B. D. Nageswara, 176, 279; 177, 358 Rao, D. N. Ramakrishna, 186, 318 Rapoport, Micha, 178, 300 Rappuoli, Rino, 178, 404 Raschke, Klaus, 174, 312 Rasmussen, Dennis D., 168, 206 Rasmussen, Howard, 191, 661 Raths, Susan, 194, 697 Rau6, H. A., 194, 453 Raven, J. A., 174, 366 Raychaudhuri, Syamal, 178, 3 Raymond, Christopher K., 194, 644 Razin, Ehud, 187, 514 Read, Marjorie S., 169, 149 Reagan, Kevin J., 184, 556 Rearick. James 1., 190, 42 Rector, Edward S., 184, 111 Reddanna, Pallu, 187, 268 Reddy, C. Channa, 187, 268 Reddy, Ram, 180, 521 Redeuilh, G6rard, 184, 292 Redfern, Christopher P. F., 189, 239, 307 Redhead, Christopher, 191, 572 Reenstra, W, W., 192, 151 Reeve, Joseph R., Jr., 168, 660 Regenass, M., 172, 528 Regnier, Evelyne, 170, 14 Regnier, Fred E., 182, 392 Rehm, Warren S., 171, 607 Reichner, Jonathan S., 179, 82 Reid, Gordon P., 172, 288 Reilly, J. David, 180, 177 Reisfeld, Avi, 184, 608 Reiss, Gloria, 183, 3 Reithmeier, Reinhart A. F., 173, 494 Renz, Mark E., 185, 421 Repine, John E., 186, 651 Reshkin, S., 191, 494

305

Retief, J. D., 170, 431 Reuss. L., 192, 650 Reyes, Vicente M., 180, 63 Rhee, Sue Goo, 186, 478 Rhodes, D., 170, 420, 575 Rhodes, David G., 182, 555,566 Rial, Eduardo, 174, 85 Rice, John, 183, 3 Rich, Alexander, 184, 328 Richards, Frederic M., 184, 3 Richter, Wilhelm J., 193, 607 Rickles, Richard J., 190, 110 Riezman, Howard, 194, 697 Rightsel, Wilton A., 191, 152 Rimmelzwaan, G. F., 178, 375 Rinck, G., 190, 291 Rine, Jasper, 194, 239 Risley, John M., 177, 376 Rivier, Catherine, 168, 588 Rivier, Jean, 168, 588 Robberecht, Wim, 168, 47 Roberts, Christopher J., 194, 644 Roberts, Gerald D., 193, 501 Roberts, L. Jackson, II, 187, 1, 51 Robertson, Hugh D., 180, 130, 418; 181, 189 Robins, Sander J., 187, 195 Rockmill, Beth, 194, 146 Roder, Heinrich, 176, 446 Roeder, G. Shirleen, 194, 146 Roelofsen, Ben, 173, 223 Roepstorff, Peter, 193, 432 Rogers, Wade T., 168, 808 Rokach, Joshua, 187, I Rol|man, Ola, 190, 163 Roop, Dennis R., 190, 91 Rosbash, M., 181, 122 R6sch, Paul, 176, 342 Rose, Alan B., 185, 234 Rose, Mark D., 194, 195 Rose, Mary Callaghan, 179, 3 Rosen, Gerald M., 186, 611 Rosenberg, Alan H., 185, 60 Rosenberg, Robert D., 169, 342 Rosenberg, Steven, 185, 341 Rosenbusch, J. P., 172, 528 Rosenthale, Marvin E., 190, 346 Rosevear, Paul R., 177, 333 Ross, A. Catharine, 189, 81,442; 190, 259 Ross. Alberta B., 186, 113

306

CONTRIBUTOR INDEX

Ross, Jeffrey, 181, 202 Rossi, Claudio, 176, 184 Rossomando, Edward F., 182, 38, 309 Rothblatt, Jonathan, 194, 662 Rothenberg, Jeffrey M., 184, 208, 608 Rothstein, Aser, 173, 401 Rothstein, Rodney, 194, 281 Rottenberg, Hagai, 171, 364; 172, 63 Roussel-Velge, Florence, 178, 390 Roux, Kenneth H., 178, 130 Rouzer, Carol A., 187, 312 Ruby, S. W., 181, 97 Rucinski, Boguslaw, 169, 22 Ruf, Horst, 172, 364 Ruggeri, Zaverio M., 169, 244 Rumrich, Gerhard, 191, 98 Runge, Marschall S., 178, 515 Runyan, Keith R., 171, 164 Ruoho, Arnold E., 184, 660 Rush, J. D., 186, 148 Ruskin, Barbara, 181, 180 Russell, Christopher B., 177, 44 Ryals, Julie, 183, 3 Ryan, Clarence A., 179, 566 Rymond, B. C., 181, 122

Saari, John C., 190, 156 Saboli6, Ivan, 191, 505 Saccone, Cecilia, 183, 570 Sachs, Alan B., 181, 332 Sachs, G., 171, 3; 191, 469 Sachs, John R., 173, 80 Sacks, Peter G., 190, 100 Saelinger, Catharine B., 184, 379 Saint-Remy, Jean-Marie R., 178, 150 Saitou, Naruya, 183, 584 Sakariassen, Kjell S., 169, 37 Sakena, Puspha, 189, 315 Sakon, Masato, 169, 442 Sala, Angelo, 187, 90 Salazar, Margarita D., 183, 3 ~ali, Andrej, 183, 670 Salustri, Antonietta, 179,435 Salyan, Mary Ellen K., 179, 242 Salzman, Edwin W., 169, 104, 386 Sammar, Marei, 178, 714 Samokyszyn, Victor M., 186, 457; 190, 281

Samuelsson, Bengt, 187, 1,296, 312,486, 491 Samuelsson, Bo E., 193, 623 Sanders, Charles R., II, 177, 317 Sanders, Dale, 174, 443 Sanders, Martin J., 171, 482 Sani, Brahma P., 189, 43, 348 Sankoff, David, 183, 428 Saran, Manfred, 186, 343 Saris, Nils-Erik Leo, 174, 68 Sasaki, Takeshi, 178, 422 Sasaki, Terukatsu, 179, 559 Sato, Fusako, 192, 583 Sato, Kenzo, 192, 583 Sauer, N., 174, 390 Sauron, M. E., 178, 144 Sauvaget, Isabelle, 183, 237 Scarborough, Gene A., 174, 667 Scarpa, Antonio, 173, 662 Schachter, Harry, 179, 351 Schafer, James A., 191, 232, 354 Schaich, K. M., 186, 121 Schechter, Yoram, 178, 300 Scheek, R. M., 177, 204 Scheffery, Carl, 171, 792 Schekman, Randy, 194, 662 Schena, Mark, 194, 389 Scherf, Christine, 194, 169 Schick, Michael R., 178, 36 Schild, David, 194, 827 Schindler, Hansgeorg, 171, 225 Schleich, Thomas, 176, 386, 418 Schlessinger, David, 181, 355 Schlessinger, Joseph, 168, 481 Schloeder, Diane, 178, 531 Schlondorff, Detlef, 191, 130 Schmaier, Alvin H., 169, 276 Schnaar, Ronald L., 179, 542 Schneerson, Rachel, 184, 537 Schneider, Barbara G., 184, 370 Schneider, Jane C., 194, 373 Schneider, Jerry A., 174, 154 Sch6neich, Jerzy T., 181, 307 Schoner, Brigitte E., 185, 94 Schoner, Ronald G., 185, 94 Schoolnik, Gary K., 178, 163 Schram, Karl H., 193, 791 Schuchmann, Heinz-Peter, 186, 511 Schultz, Loren D., 185, 297 Schultz, Patrick, 170, 166

CONTRIBUTOR INDEX Schultz, Peter G., 178, 551 Schulz, Irene, 192, 280 Schulz, John T., 173, 676 Schulz, Riidiger, 178, 243 Schiimperli, Daniel, 181, 74 Schuricht, B., 191, 437 Schwab, A., 191, 437 Schwaber, James S., 168, 778, 808 Schwager, S., 170, 431 Schwartz, Ariela, 178, 714 Schwartz, George J., 191, 253 Schwartz, Jeffrey, 168, 29 Schwartz, M., 171, 607 Schwartzman, Michal Laniado, 187, 372 Schwarz, Alexander, 184, 160 Schwarz, W., 173, 112,453 Schweikl, Helmut, 192, 608 Schwoebel, Eric D., 182, 663 Scoble, Hubert A., 193, 519 Scott, Carolyn D., 168, 309 Scotto, Anthony W., 171, 253 Seabold, Robert R., 187, 175 Sealy, Linda, 170, 612 Secco, Claude, 184, 292 Segal, Y., 192, 650 Seibicke, Stefan, 191, 583 Seifert, H. Steven, 194, 329 Seifter, Sam, 182, 285,626 Semenza, G., 192, 409 Senapathy, Periannan, 183, 252 Senior, Robert M., 169, 233 Seraphin, B., 181, 122 Serbinova, Elena E., 186, 355 Serhan, Charles N., 187, 167 Sewell, B. T., 170, 431 Seyama, Yousuke, 187, 286, 296, 486 Shafik, lhab M., 191, 72 Shakhov, A. N., 170, 408 Shaltiel, Shmuel, 186, 464 Shao, Ming-Chuan, 184, 653 Shapiro, Marvin B., 183, 252 Shapiro, Stanley S., 190, 326, 334, 352 Shatkin, Aaron J., 180, 164 Shattil, Sanford J., 169, 355 Shaw, Gary S., 189, 112 Shearman, Mark S., 168, 347 Sheldon, Edward L., 184, 577 Shen, T. Y., 187, 446 Shepherd, John C. W., 183, 180 Sherman, Fred, 194, 3, 21, 38, 362

307

Sherman, Mark A., 178, 91 Sherman, Michael 1., 189, 337, 525; 190, 148 Shero, James H., 194, 749 Sheshberadaran, Hooshmand, 178, 746 Shick, V. V., 170, 386 Shigenaga, Mark, 186, 521 Shimamura, Akiko, 170, 603 Shimizu, Takao, 187, 286, 296, 486 Shimura, Yoshiro, 181, 209, 395 Shin, Seung-Uon, 178, 459 Shiraishi, Hideaki, 181, 395 Shirley, Michael A., 187, 277 Shively, John E., 184, 507, 518; 193, 361 Shiver, J. W., 172, 439 Shuman, H., 172, 203 Shuman, Stewart, 181, 170 Siebert, Friedrich, 189, 123 Siegenthaler, Georges, 189, 299, 530 Sies, Helmut, 173, 523; 174, 118; 186, 595 Sikorski. Robert S., 194, 302 Silberstein, Leslie E., 179, 299 Silbert, Jeremiah E., 179, 422,428 Silhavy, Thomas J., 185, 166 Silva, Patricio, 191, 31; 192, 754 Silverman, Melvin, 191, 34 Simasko, Steven M., 192, 309 Simchen, Giora, 194, 94 Simic, Michael G., 186, 89 Simmons, N. L., 191, 426 Simon, Ethan S., 179, 275 Simons, Kai, 191, 813 Simonson, Michael S., 187, 544 Simpson, K. L., 189, 433; 190, 237 Simpson, Nancy J., 185, 421,471 Simpson, Robert T., 170, 26 Singer, Birgit, 194, 697 Singer, S. J., 184, 343 Singh, Ajit, 186, 686 Singh, Arjun, 185, 421 Singh, Harwant, 186, 686 Sirois, Pierre, 187, 98 Siu, Chi-Hung, 189, 356 Sixma, Jan J., 169, 301 Sjodin, Raymond A., 173, 695 Skerra, Arne, 178, 497 Skingle, Derek C., 184, 588 Skinner, Sheri M., 182, 670 Skorecki, Karl, 191, 551 Skutelsky, Ehud, 184, 340

308

CONTRIBUTOR INDEX

Slayman, Clifford L., 174, 654 Slayter, Henry S., 169, 251,326 Sledziewski, Andrzej Z., 185, 351 Slot, Jan-Willem, 169, 301 Smigel, Murray D., 172, 696 Smith, David F., 179, 30 Smith, David L., 193, 374 Smith, John Edgar, 190, 229 Smith, Laura L., 178, 693 Smith, Leland L., 186, 438 Smith, Margaret L., 174, 154 Smith, Martyn T., 186, 579 Smith, Richard D., 193, 412 Smith, William L., 171, 581; 187, 1,469 Snavely, Marshall D., 173, 546 Soberman, Roy J., 187, 319, 335 Soboll, Sibylle, 174, 118 SOderstrOm, Mats, 187, 306 Sogo, Jos6 M., 170, 142 Sokolov, A. P., 188, 339 Soldati, Dominique, 181, 74 Soil, Andrew H., 171, 482 S011, Dieter, 181, 377, 451 Solomon, Arthur K., 173, 192 Solomon, Richard J., 192, 754 Somlyo, A. P., 172, 203 Somlyo, A. V., 172, 203 Sommer, Andreas, 185, 187 Sone, Masayoshi, 191, 409 Sonnenburg, William K., 171, 581 Soprano, Dianne Robert, 189, 286 Sottocasa, Gian Luigi, 174, 50 Souroujon, Miriam C., 178, 433 Spaeth, Daniel D., 192, 599 Spanswick, Roger M., 174, 479 Sparks, Steven W., 176, 134 Speilman, William S., 171, 581 Spencer, Forrest, 194, 749 Spillmann, Dorothe, 179, 270 Spitalnik, Steven L., 179, 299 Sprague, George F., Jr., 194, 77, 187 Srere, Paul A., 182, 539 Srivastava, Anand K., 181, 355 Stadelmann, Eduard J., 174, 225,246 Staden, Rodger, 183, 163, 193 Stader, Joan A., 185, 166 Stadtman, Earl R., 182, 793; 186, 464, 478 Stafforini, Diana M., 187, 344 Stampfer, Meir J., 189, 155 Stanway, C. A., 185, 329

Stark, G., 171, 274 Staros, James V., 172, 609, 687 Stauber, Claudia, 181, 74 Stearns, Tim, 185, 280 Steck, Theodore L., 173, 356, 513 Steele, Roderick E., 171, 736 Stein, Arnold, 170,585 Stein, Evan A., 189, 155 Stein, Wilfred D., 171, 23 Steiner, Melissa Gale, 186, 137 Steipe, Boris, 182, 132 Steitz, Joan A., 180, 454, 468 Stellwagen, Earle, 182, 317, 343 Stelts, Stephen R., 183, 3 Stene, Danny O., 187, 277 Stephens, Robert J., 186, 388, 399 Steudle, Ernst, 174, 183 Stevens, Fred J., 178, 107 Stevens, Tom H., 194, 644 Stevenson, George T., 178, 341 Stewart, James R., 186, 742 Stewart, P. L., 176, 242 Stieger, Bruno, 172, 346 Stiekema, Frank, 184, 314 Stillwell, William, 189, 373,383 Stitt, Mark, 174, 518 Stocker, Roland, 186, 301 Stocum, David L., 190, 189 Stoll, Vincent S., 182, 24 Stone, Kathryn L., 193, 389 Stormo, Gary D., 183, 211; 185, 89 Storrie, Brian, 182, 203 Stoscheck, Christa M., 181, 50 Stotz, Agathe, 194, 169 Stoytchev, Tsanko S., 186, 355 Strasburger, Christian J., 184, 481 Strathern, Jeffrey N., 194, 319, 342 Strauss, Ellen G., 180, 121 Strauss, James H., 180, 121 Strickland, Sidney, 190, 110 Strosberg, A. D., 178, 179, 265 Struhl, Kevin, 194, 520 Stryer, Lubert, 184, 188 Studier, F. William, 185, 60 Stults, Cheryl L. M., 179, 167 Stump, W. Thomas, 187, 175 Sugumaran, Geetha, 179, 422, 428 Sumida, Satoshi, 186, 197 Sussman, Joel L., 184, 90 Sussman, Marc S., 186, 711

CONTRIBUTOR INDEX Sutoh, Kazuo, 184, 409 Sutton, Ann, 184, 537 Suzuki, Koichi, 187, 486 Suzuki, Tsuneo, 179, 331 Swank-Hill, Patti, 179, 542 Swanson, Maurice S., 181, 326 Swearingen, Karen C., 168, 254 Sweeley, Charles C., 179, 167 Swennen, Luc, 168, 47 Symons, Robert H., 181, 583; 184, 588 Sytkowski, Arthur J., 184, 353

T Takagi, Toshio, 172, 514 Takahashi, Mark, 192, 438 Takahashi, Noriko, 189, 233 Takashima, Ikuo, 184, 497 Tam, James P., 168, 7; 178, 739 Tanaka, Atsuo, 188, 171, 176, 463 Tang, Laura, 192, 124 Taniguchi, Naoyuki, 179, 397, 570 Tanner, Michael J. A., 173, 423 Tanner, N. Kyle, 180, 25 Tanner, W., 174, 390 Tarara, Thomas E., 189, 418 Taylor, Paul B., 193, 501 Taylor, Wayne E., 185, 308 Taylor, William R., 183, 456 Teelmann, Kampe, 190, 382 Tekamp-Olson, Patricia, 185, 341 Terada, Lance S., 186, 651 Ternynck, Th6r6se, 184, 469 Tesoriere, Luisa, 189, 369, 503 Thaller, Christina, 190, 201 Th6ze, J., 178, 144 Thom, Margaret, 174, 552 Thoma, Fritz, 170, 142 Thomas, David W., 186, 388, 399 Thomas, Jean O., 170, 369, 549 Thomas, John O., 181, 307 Thomas, Thomas C., 182, 499 Thomason, J. F., 177, 159 Thompson, Robert C., 185, 187 Thor, Hj6rdis, 186, 627 Thornton, George B., 178, 634 Thorsness, Peter E., 194, 149 Thiirich, T., 186, 723 Tien, Ming, 188, 159

309

Timmers, Adrian M., 190, 3 Timmons, Sheila, 169, 11 Timmons, Therese M., 182, 441,679 Tinoco, Ignacio, Jr., 180, 304 Tiribelli, Claudio, 174, 50 Tocchini-Valentini, G. P., 181, 510 Tomlinson, Laurie J., 183, 3 Toone, Eric J., 179, 275 Torchia, Dennis A., 176, 134 T6rm~, Hans, 190, 163,210 Torres, B. Virginia, 179, 30 Touati, Daniele, 186, 646 Touster, Oscar, 179, 446, 500 Townsend, R. Reid, 179, 65 Tramontano, Alfonso, 178, 531 Trapane, T. L., 171, 286 Trentham, David R., 172, 288 Tridente, Giuseppe, 178, 404 Trotsenko, Y. A., 188, 339 Troyer, D. A., 191, 141 Trudgill, Peter W., 188, 70, 77 Trueheart, Joshua, 194, 774 Trujillo, Brian L., 183, 3 Tsai, Ming-Daw, 177, 317 Tsien, Roger, 172, 230 Tsolas, O., 182, 788 Tsygankov, Yuri D., 188, 247, 335 Tullius, Thomas D., 186, 545 Tulsiani, Daulat Ram P., 179, 446, 500 Turk, John, 187, 175 Turner, Douglas H., 183, 281 Turner, R. James, 191, 479; 192, 26 Twining, Sally S., 190, 259 Tzagoloff, Alexander, 194, 477 Tzukerman, Mary, 189, 256

U Ueda, Mitsuyoshi, 188, 171, 176, 463 Ueda, Natsuo, 187, 338 Ueda, Tetsufumi, 174, 9 Uhl, George R., 168, 741 Uhl6n, Mathias, 185, 129 Uhlenbeck, Olke C., 180, 51 Ullman, M. D., 172, 538 Ullrich, Karl Julius, 191, 1, 98 Umetsu, Kazuo, 179, 331 Underwood, Barbara A., 190, 242 Urry, Dan W., 171, 286

310

CONTRIBUTOR INDEX

Ursini, Fulvio, 186, 448 Uster, Paul S., 171, 850 UytdeHaag, F. G. C. M., 178, 375

V Vadgama, Jaydutt V., 171, 133 Vahlquist, Anders, 190, 163, 210 Vaidya, Chitra M., 187, 406 Vale, Wylie, 168, 29, 588 Valiquette, Guy, 168, 574 Vallerand, Pierre, 187, 98 van Adelsberg, Janet, 172, 85 van Corven, Emile J. J. M., 192, 341 van der Klei, Ida J., 188, 411,420 van der Meer, R. A., 188, 260 Vanderwel, Marianne, 169, 455 Van Etten, Robert L., 177, 376 van Gunsteren, W. F., 177, 204 Vanhove, A., 187, 70 van Kleef, M. A. G., 188, 260 van Kuijk, Frederik J. G. M., 186, 388, 399 van Lier, J. E., 186, 502 Vann, Willie F., 184, 537 Van Regenmortel, Marc H. V., 170, 251 Van Tol, Hubert H. M., 168, 398 van Wielink, John E., 188, 235 Varki, Ajit, 179, 409, 416 Vath, James E., 193, 738 Vaughan, Joan M., 168, 588 Vaz, Alfin D. N., 186, 278 Vaz, Winchel L. C., 172, 471 Veenhuis, Marten, 188, 411 Venkatachalam, C. M., 171, 286 Venton, Duane, 187, 245,406 Verkman, Alan, 191, 551 Vestal, Marvin L., 193, 107 Vicente, Oscar, 181, 499 Vigan6, T., 187, 621 Vihinen, Mauno, 183, 447 Villafranca, Joseph J., 177, 390, 403 Vinayek, Rakesh, 191, 609 Vingron, Martin, 183, 352 Vioque, Agustin, 181, 569 Viscidi, Raphael P., 184, 600 Vishkin, Uzi, 183, 487 Vlodavsky, Israel, 169, 76 Voelkel, Norbert F., 187, 599

Vogel, Hans J., 177, 263 Voglmayr, Josef K., 168, 588 von Dippe, Patricia, 174, 25 von Holt, C., 170, 431 von Sonntag, Clemens, 186, 511 Vrbanac, J. James, 187, 62

W

Wackett, Lawrence P., 188, 39 Wagener, Christoph, 184, 507, 518 Wagner, Gerhard, 176, 93 Wagner, J. R., 186, 502 Wakem, Paul, 194, 38 Wakui, M., 192, 300 Walker, Jeffrey W., 172, 288 Walker, Scott S., 182, 521 Wallace, James C., 183, 111 Wallace, John C., 180, 177 Wallmark, BjOrn, 191, 721 Walsh, John H., 168, 660 Walter, Peter, 194, 675 Waiters, Julian R. F., 192, 448 Wandinger-Ness, Angela, 191, 813 Wandrey, Christian, 184, 160 Wang, Kuan, 172, 687 Wang, Paulus S., 168, 371 Wang, W., 179, 55 Wang, W., 192, 710 Wank, Stephen A., 192, 247 Ware, J. Anthony, 169, 386 Warner, Jonathan R., 194, 423 Warnke, Roger A., 184, 363 Waseem, Ahmad, 173, 513 Wassail, Stephen R., 189, 373,383 Wassarman, Paul M., 170, 290 Waterman, Michael S., 183, 221 Watson, J. Throck, 193, 86 Watson, Robert, 184, 577 Wedden, Sarah, 190, 201 Weerheim, Arij, 19tl, 30 Wehner, F., 192, 650 Wehrly, John A., 184, 556 Weiner, David B., 178, 321 Weiner, Mark G., 183, 3 Weiner, Richard I., 168, 254 Weiser, Milton M., 192, 448 Weisleder, D., 186, 380 Weisman, Gary A., 171, 857

CONTRIBUTOR INDEX Weiss, Etienne, 170, 14 Weiss, George H., 176, 3 Weiss, Lawrence M., 184, 363 Weiss, Shelly, 182, 371 Welch, Frank J., 183, 3 Wells, John, 171, 497 Wendel, Albrecht, 186, 675 Wendelborn, Daniel F., 187, 51 Werdan, Karl, 173, 634 Werner, G., 178, 676 Westphale, H.-J., 191, 437 Wetsel, W., 168, 517 Whatley, Ralph E., 187, 520 Whelan, J., 187, 268 Whitaker, John N., 178, 63 White, Jeffrey D., 168, 681 White, Ronald E., 186, 273 White, Stephen, 182, 646 Whitehead, James S., 184, 201 Whiteheart, Sidney W., 179, 82 Whitesides, George M., 179, 275 Whittaker, James W., 188, 82 Wickens, Marvin, 180, 369 Wickert, Lucia, 184, 507 Widdas, W. F., 173, 231 Widger, W. R., 172, 439 Wiebauer, Karin, 181, 148 Wieczorek, Helmut, 192, 608 Wiederholt, Michael, 192, 571 Wiemken, Andres, 174, 504 Wiggert, Barbara, 189, 213 Wiig, Susan E., 183, 3 Wilchek, Meir, 184, 5, 14, 49, 68, 80, 123, 138, 160, 174, 208, 217, 243,301, 340, 415,427,467, 560, 608, 618 Wilde, Joyce A., 177, 74, 282 Wilkinson, Keith D., 185, 387 Willhite, Calvin C., 190, 406 Williams, David E., 187, 253 Williams, G. Melville, 186, 748 Williams, Jacinta B., 181, 451 Williams, James C., Jr., 191, 232, 354 Williams, John T., 168, 103 Williams, Kenneth R., 193, 389 Williams, Patrick L., 183, 615 Williams, William V., 178, 321 Williamson, John R., 191, 691 Willis, Anthony L., 187, 1 Willis, Ian, 181, 377 Wills, Ken N., 189, 256

311

Wills, Nancy K., 171, 715 Wilson, M., 185, 329 Wilson, William E., 168, 444 Winguth, Sandra D., 182, 688 Winicov, Irene, 168, 218 Winiski, Anthony, 171, 342 Winkler-Oswatitsch, Ruthild, 183, 505 Winterbourn, Christine C., 186, 265 Wirthensohn, Gabriele, 191, 325 Wirthmueller, Urs, 187, 567 Wise, Jo Ann, 194, 405 Wohlhueter, Robert M., 173, 714 Wold, Finn, 184, 653 Wolf, Barry, 184, 103 Wolf, Bryan A., 187, 175 Wolfersberger, Michael G., 192, 608 Wolgel, Sanford A., 188, 95 Wollheim, Claes B., 192, 188, 223 Wood, Phillip G., 173, 346 Woodcock, Christopher L. F., 170, 180 Woodle, Martin C., 171, 193 Worcel, Abraham, 170, 603 Wormmeester, Jan, 184, 314 Worthington, Marlis, 185, 308 Wozney, John M., 182, 738 Wreggett, K. A., 191, 707 Wright, P. A., 191, 226 Wright, Peter E., 176, 114 Wu, Carl, 170, 269 Wu, Kenneth K., 187, 578 Wfithrich, Kurt, 177, 125 Wyss, Ronald, 189, 146

X Xin, Lu, 178, 212 Xu, Qi, 181, 463

Y Yadagiri, Pendri, 187, 357 Yaffe, Michael P., 194, 627 Yalow, Rosalyn S., 168, 302 Yamada, Ryo, 185, 398 Yamada, Tadataka, 192, 176 Yamagata, Tatsuya, 179, 488 Yamamoto, Keith R., 194, 389 Yamamoto, Shozo, 187, 1, 338

312

CONTRIBUTOR INDEX

Yamamoto, Yorihiro, 186, 371 Yamashina, Ikuo, 179, 310, 322 Yamashiro, Carl T., 194, 644 Yamashita, Katsuko, 179, 46, 331 Yanagishita, Masaki, 179,435 Yankaskas,James R., 192, 565 Yansura, Daniel G., 185, 54, 161 Yared, Aida, 191, 107 Yeo, Erik L., 169, 311 Yip, Carli, 185, 351 Yisraeli, Joel K., 180, 42 Yolken, Robert H., 184, 529 Yoshigaga, Kaoru, 178, 422 Yoshikawa, Toshikazu, 186, 656, 660 Yost, Richard A., 193, 154 Youdim, Moussa B. H., 178, 308 Youle, Richard J., 178, 356 Young, Elton T., 185, 308 Young, Richard A., 194, 230, 508 Young, W. Scott, III, 168, 702

Yudin, Katherine, 183, 3 Yule, D., 192, 300

Z Zakim, David, 171, 253 Zaug, Arthur J., 181, 558 Zhang, Xiao-kun, 189, 256 Zhou, Zhongrui, 193, 374 Ziegler, D. M., 186, 621 Zimmer, G., 186, 723 Zimmerman, Guy A., 187, 520 Zimmerman, Theodore S., 169, 244 Zimmermann, U., 174, 338 Zins, Larry B., 183, 3 Zopf, D., 179, 55 Zucker, Marjorie B., 169, 117 Zuckier, Gerald N., 174, 654 Zuker, Michael, 180, 262; 183, 281 Zulauf, M., 172, 528