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This volume of Methods in Enzymology and its companion Volume 238 include molecular, biological, and biochemical methods for the study of cell surface heterotrimeric G proteins (Volume 237) and their effectors (Volume 238). Methods unique to signal transducing G proteins and general techniques applied to the study of G protein systems are covered.
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This book is printed on acid-free paper. e Copyright © 1994 by ACADEMIC PRESS. INC. All Rights Reserved. No pan of this publkation may be reproduced or transmitted in any form or by any means. electronic or mechanical. including photocopy. recording. or any information storage and retrieval system. without permission in writing from the publisher. Academic Press, Inc. 525 B Street. Suite 1900. San Diego. California 92101-4495 United Kingdom Editio/I published by Academic Press Limited 24-28 Oval Road. London NW I 7DX International Standard Serial Number: 0076-6879 International Standard Book Number: 0·12-182138-2 PRINnD IN mE UNITED STATFS OF AMERICA 94 95 96 97 98 99 MM 9 8 7 6 5 4 3 2 I Table of Contents CONTRI8UTORS TO VOLUME 237 . PRt:f"CE . VOLUMES IN SERIES )1'111 Section I. Ga Subunits I. Measurement of Receptor-Stimul"ted Guanosine THOM"S Wlt:I.AND AND Y-O'(y,ThiO)triphosphate Binding by G Pro- K A RL H. JAKOHS teins 2. Receptor-Slimulated Hydrolysis of Guanosine 5' PETER GIERSCIUK. Triphosphate in Membra.ne PreparJ.tions THOMAS BOUILLON. AND K"II.L H. JAKOHS 3. Regulation of G-Protein Activation by Mastopar-lOs and Other Clilionic Pcptides 13 EI.I.IOTT M. Ross AN\} 26 TSUTOMU HIG"SHIJIM" 4. Guanosine 5'-O-(y·Thio)triphosphate Binding As- DoNNA 1. CARTY "NO say for Solubilized G PrOleins J8 R"vt IYENGAR 5. Activation ofCholem TO)lin by ADP-Ribosylation JOEL M oss . Factors: 20-kDa Guanine Nucleotide.Binding R"NOY S. H"UN. Proteins SU CI IEN TSA1, , CATHEII.INE F. Wnsli. FANG-JEN SCOTT LEE, S. Russ PRICE. AND 6. Pertussis Toxin-Catalyzed ADP·Ribosylation of 0 MARTHA VAUGHAN 44 DoNNA 1. CARTY 63 Proteins 7. Synthesis and Use of Radioactive Pholoaclivat- R1CIIARD R. VAILLANCOURT. able NAO' Derivatives as Probes forG-Protein N. DIIANASEKARAN. AND Structure ARNOLD E. Ruouo 8, PholoaffinilY Guanosine Y-Triphosphate Analogs MARK M. RAS EN1CK 70 . as a Tool for the Study ofGTP-Binding Proteins MADIIAVI TAI.I.UIU. AND WILLIAM J. DUNN III 9. Prepar.l.tion of Activated a Subunits ofG, andG,s: From Erythrocyte to Activated Subunit 100 LUTZ BIRNHAUMER, D"GOHERTO GRENET. FERNANDO RI8EIRO-NETO. AND JUAN CODINA 110 TABLE OF COI"'TE",TS v, 10. l>Urification and Separation of Closely Related TOSHI""I KAT"!),,, Membe� of Pertussis Toxin-Substrate G Pro· KENJI KONTANI. teins ATSUSHI INANORF-. [CHIRQ KOBAYASHI. YOSHIIIARU OIlOK". HIItOSIil NISltlNA. AND KATSUNOHU TAKAHASHI II. Purification of Tmnsducin 131 JOf!:LLE BIGA\' AND MARC (HARRE 12. Expression orO-Protein n Subunits in £S{'/lI'ridri" '39 ETUAN LEE. MAURINE E. LINDI,It, AND coli Al.FRED G. GII.MAN '4{\ 13. Synthesis and Applications of Affinity Matrix IOK-Hou PAf"G. Containing Immobilized (ly Subunits orG Pro- ALAN V. SM lleK" , 14. Purification "I'll) PAUL C. STERNWEIS teins of Activated and 1