Table Of ContentMethods in Enzymology
Volume 98
Biomembranes
Part L
Membrane Biogenesis: Processing and Recycling
EDITED BY
Sidney Fleischer
BeccaF leischer
DEPARTMENT OF MOLECULAR BIOLOGY
VANDERBILT UNIVERSITY
NASHVILLE, TENNESSEE
Editorial Advisory Board
David Baltimore Walter Neupert
Giinter Blobel George Palade
Nam-Hai Chua David Sabatini
1983
ACADEMIC PRESS
A Subsidiary ofHarcourt Brace Jovanovich, Publishers
New York London
Paris San Diego San Francisco Slo Paulo Sydney Tokyo Toronto
Contributors to Volume 98
elcitrA numbers are in parentheses gniwollof the names of contributors.
snoitailiffA listed are current.
PETER ARVAN (8), Section of Cell Biology, cultk de M(dicine, F 13385 Marseille
Yale ytisrevinU, School of Medicine, New Cedex ,5 France
Haven, Connecticut 06510 A. M. NIR~uG-TUABMAHC (15), Laboratoire
KIBOOR AZARNIA (47), Department of Physi- de Biochimie des Transports Cellulaires,
ology and Biophysics, University of Miami, Institut de Biochimie, ~tisrevinU Paris Sud,
Miami, Florida 33101 91405 Orsay Cedex, France
JACQUES U. BAENZIGER (12), tnemtrapeD of LIVNAZ A. COHN (35), Department of -ulleC
Pathology, Washington University School lar Physiology and Immunology, The
of Medicine, St. Louis, Missouri 63110 Rockefeller University, New York, New
P. F. BAKER (3), Department of ,ygoloisyhP York 10021
King's College London, Strand, London JERRY R. COLCA (16), Department of -lohtaP
WC2R 2LS, England ogy, Washington University School of Med-
WILLIAM E. BALCH (4), Department of -DiB ,enici St. Louis, Missouri 63110
chemistry, Stanford University, Stanford, IOM E. CREEK (23), The Edward Mal-
ainr~lfilaC 94305 linckrodt Department of Pediatrics, Wash-
PIDNAS K. BASU (19), Department of -uceloM ington University School of Medicine, and
lar Genetics, University of Texas Health Division of Medical Genetics, St. Louis
Science Center at Dallas, Dallas, Texas Children's Hospital, St. Louis, Missouri
;"3257 63178
LABUS EEYAHSIB (49), Department of Bio- GERHARD DAHL (47), Department of Physi-
chemistry and Biophysics, University of ology and Biophysics, University of Miami,
Pennsylvania School of Medicine, Philadel- Miami, Florida 33101
phia, Pennsylvania 19104 JOHN L. DAISS (29), Biosciences Division,
GONTER LEBOLB (40), Laboratory of Cell Bi- Kodak Research Laboratories, Eastman
ology, The Rockefeller University, New Kodak Company, Rochester, New York
York, New York 10021 14650
~3JANREB BLOJ (51), Division of Nutritional MANJUSRI SAD (49), Department of Bio-
Sciences and Section of Biochemistry, Mo- chemistry and Biophysics, University of
lecuktr, and Cell Biology, Cornell Univer- Pennsylvania School of Medicine, Philadel-
sity, Ithaca, New York 14853 phia, Pennsylvania 19104
EUQIRNE NADNARB (28), Departamento Bio- RANDALL L. DIMOND (13), Department of
logia Celular, Universidad Catolica, Cas- Bacteriology, University of Wisconsin,
silla 114-D, Santiago, Chile Madison, Wisconsin 53706
LEAHCIM .S NWORB (19), Department of Mo- JACK J. DISTLER (25), Rackham Arthritis Re-
r~hucel Genetics, University of Texas search Unit and Department of Internal
Health Science Center at Dallas, Dallas, Medicine, University of Michigan, Ann
Texas 75235 Arbor, Michigan 48109
RICHARD S. CAMERON (8), Section of Cell MAILLIW A. DUNN (18), Department of Cell
Biology, Yale University School of Medi- Biology and Anatomy, The Johns Hopkins
cine, New Haven, Connecticut 06510 University School of Medicine, Baltimore,
J. DAVO CASTLE (8), Section of Cell ,ygoloiB Maryland 21205
Yale University School of Medicine, New WILLIAM G. DUNPHY (4), Department of
Haw,n, Connecticut 06510 Biochemistry, Stanford University, Stan-
ENNAIRAM DRABMAHC (42), Laboratoire de ford California 94305
Biochimie elacidOM et INSERM U38, Fa- HANSJORG EIBL (58), Max-Planck-Institut
xi
xii CONTRIBUTORS TO VOLUME 98
fftr Biophysikalische Chemie, Karl-Fried- 1RA HELENIUS (20), Section of Cell Biology,
rich-Bonhoeffer-lnstitut, Abteilung ,31 Yale University School of Medicine, New
D-3400 GOttingen-Nikolausberg, Federal Haven, Connecticut 06510
Republic of Germany ELLIOT L. HERTZBERG (44), Verna and
ALAN D. ELBEIN (11), Department of Bio- Marrs McLean Department of Biochemis-
chemistry, University of Texas Health try, Baylor College of Medicine, Texas
Science Center, San Antonio, Texas 78284 Medical Center, Houston, Texas 77030
MARILYN GIST FARQUHAR (1, )71 Section REKLOV HERZOG (17, 39), Institutfiir Zell-
of Cell Biology, Yale University School biologie der tigtisrevinU ,nehcn2fM D-8000
of Medicine, New Haven, Connecticut Mfmchen, Federal Republic of Germany
06510 TREBOR L. HILL (26), Department of Bio-
JEFFREY FEINMAN (49), Department of -DiB chemistry, Duke University Medical
chemistry and Biophysics, University of Center, Durham, North Carolina 27710
Pennsylvania School of Medicine, Philadel- G. HOLT (24), Department of Physiology and
phia, Pennsylvania 19104 Biophysics, Washington University School
n. DAVID REHCSIF (23), Department of -DiB of Medicine, St. Louis, Missouri 63110
chemistry, Molecular Biology, and Cell Bi- ANN L. HUBBARD (18), Department of Cell
ology, Northwestern University, Evanston, Biology and Anatomy, The Johns Hopkins
Illinois 60201 University School of Medicine, Baltimore,
EDUARD A. M. FLEER (58), Department of Maryland 21205
Molecular Biology, Vanderbilt University GEORGE W. JOURDIAN (25), Raekham Ar-
School of Medicine, Nashville, Tennessee thritis Research Unit and Department of
37232 Internal Medicine, University of Michigan,
ACCEB FLEISCHER (6, 28), Department of Ann Arbor, Michigan 48109
Molecular Biology, Vanderbilt ,ytisrevinU H. H. KAMP (52), Laboratory of Biochemis-
Nashville, Tennessee 37235 try, State University Hospital, Utrecht,
YENDIS REHCSIELF (28, 56, 58), Department NL-3500 CG Utrecht, The Netherlands
of Molecular Biology, Vanderbilt Univer- SEMAJ H. KEEN 13( ,) Fels Research Institute
sity, Nashville, Tennessee 37235 and Department of Biochemistry, Temple
ERIK FRIES (4), The Wallenberg Laboratory, University School of Medicine, Philadel-
University of ,alasppU 22 Uppsala, Sweden phia, Pennsylvania 19140
ENILEUQCAJ GAaRION (42), Laboratoire de TREBLIG A. KELLER (32), Department of Bi-
Biochimie M(dicale et INSERM U38, Fa- ology, University of California at San
Otluc de M(dicine, F 13385 Marseille Diego, La Jolla, California 92037
Cedex ,5 France EUGENE P. KENNEDY (55), Department of
AIHTNYC J. YAWOLLAG (48), noitceS of lleC Biological Chemistry, Harvard Medical
Biology, Yale University School of Medi- School, Boston, Massachusetts 02115
cine, New Haven, Connecticut 06510 DAVID A. KNECHT (13), Department of Biol-
NOTRON B. ALUL1G (45), tnemtrapeD of lleC ogy, University of California at San Diego,
Biology, Baylor College of Medicine, Texas La Jolla, California 92037
Medical Center, Houston, Texas 77030 D. E. KNIGHT (3), Department of ,ygoloisyhP
PAUL GLEESON (I0), National Institute for King's College London, Strand, London
Medical Research, The Ridgeway, London WC2R 2LS, England
NW7 1AA, England M. KONISH (24), Department of Physiology
HPESOJ L. GOLDSTEIN (19), Department of and Biophysics, Washington University
Molecular Genetics, University of Texas School of Medicine, St. Louis, Missouri
Health Science Center at Dallas, Dallas. 63110
Texas 75235 ORUSTET KONO (38), Department of -loisyhP
HARRY T. HAIGLER (22), Department of ogy, Vanderbilt University School of Medi-
Physiology and Biophysics, University of cine, Nashville, Tennessee 37232
,ainrofilaC Irvine, California 92717 ALAMRIN KOTAGAL (16), Department of -aP
CONTRIBUTORS TO VOLUME 98 xiii
thology, Washington University School of Pennsylvania School of Medicine, Philadel-
Medicine, St. Louis, Missouri 63110 phia, Pennsylvania 19104
NILAN M. KUMAR (45), Department of Cell TREBOR C. MIERENDORF, JR. (13), -trapeD
Biology, Baylor College of Medicine, Texas ment of ,ygoloiretcaB University of Wiscon-
Medical Center, Houston, Texas 77030 sin, Madison, Wisconsin 53706
PAUL E. LACY (16), Department of Pathol- DIANE C. MITCHELL (25), Rackham sitirhtrA
ogy, Washington University School of Med- Research Unit and Department of lnternal
,enici St. Louis, Missouri 63110 Medicine, University of Michigan, Ann
MARK .k/. NAMRHEL (26), Department of Mo- Arbor, Michigan 48109
lecular Genetics, University of Texas KEITH E. VOTSOM (40), Laboratory of Cell
Health Science Center at Dallas, Dallas, Biology, The Rockefeller University, New
Texas 75235 York, New York 10021
MAILLIW LENNARZ (9), Department of Bio- P. MULLER (15), Laboratoire de Biochimie
chemistry and Molecular Biology, Univer- des Transports Cellulaires, Institut de Bio-
sity of Texas System Cancer Center, M. .D chimie, Universit~ Paris Sud, 91405 Orsay
Anderson Hospital and Tumor Institute, Cedex, France
Texas Medical Center, Houston, Texas WILLIAM A. MULLER (35), Department of
77030 Cellular Physiology and Immunology, The
.lEINAD DRAVUOL (33), Laboratoire de Bio- Rockefeller University, New York, New
logie des Membranes, Institut Pasteur, York 10021
F-75724 Paris Cedex ,51 France SAROJA NARASIMHAN (10), Department of
LEAHCIM L. McDANIEL (16), tnemtrapeD of Biochemistry, Hospital for Sick Children,
Pathology, Washington University School Toronto, Ontario MSG IX8, Canada
of Medicine, St. Louis, Missouri 63110 BRUCE J. NOSLOHCIN (46), noisiviD of -loiB
J. OLreEg MCINTYRE (58), Department of ,ygo California Institute of Technology, -aP
raluc?hoM Biology, Vanderbilt University sadena, California 91125
School of Medicine, Nashville, Tennessee PAULA NORTH (56), Department of Physiol-
37232 ogy, Vanderbilt University School of Medi-
MARK MARSH (20), Section of lleC ,ygoloiB cine, Nashville, Tennessee 37232
Yale University School of Medicine, New IRA H. PASTAN (21), Laboratory of Molecu-
Haven, Connecticut 06510 lar Biology, National Cancer Institute, Na-
KARL MATLIN (20), naeporuE raluceloM -iB tional Institutes of Health, Bethesda,
ygolo Laboratory, 6900 Heidelberg, Fed- Maryland 20205
eral Republic of Germany B. M. F. ESRAEP (27), Laboratory of Molecu-
NAEJ PMAHCUAlV~ (42), Laboratoire de Bio- lar Biology, Medical Research Council
chimie M~dicale et INSERM U38, Facultd Center, University Medical School, Cam-
de ,enicidOM F 13385 Marseille Cedex ,5 bridge CB2 2QH, England
France MARK REGNETTIP (49), Department of Bio-
OPOCAJ lSELODLEM (7), Department of Phar- chemistry and Biophysics, University of
macology and CNR Center of Cytopharma- Pennsylvania School of Medicine, Philadel-
colcgy, tgtisrevinU Degli Studi di Milano, phia, Pennsylvania 19104
20129 Milano, Italy .B J. H. M. SIUHTROOP (53, 54), Laboratory
ARI NAMLLEiV/ (48), Section of Cell Biology, of Biochemistry, State University of
Yale University School of Medicine, New ,thcertU NL-3584 CH ,thcertU The Nether-
Haven, Connecticut 06510 lands
~tNOMYAR J. METZ (57), Mental Health Re- S. NIKZSUP (30), Department of Pathology,
search Institute and Department of -goloiB Mount Sinai School of Medicine, City -inU
'iaci Chemistry, University of Michigan, versity of New York, New York, New York
Ann Arbor, Michigan 48109 10029
NAMREH LEAHCIM (49), Department of Bio- NAMRON S. RADIN (57), Mental Health Re-
chemistry and Biophysics, University of search Institute and Department of -goloiB
xiv CONTRIBUTORS TO VOLUME 98
ical Chemistry, University of Michigan, F. ANTHONY SIMION (28), Department of
Ann Arbor, Michigan 48109 Molecular Biology, Vanderbilt University,
NIVLOC M. REDMAN (14), Lindsley F. Kim- Nashville, Tennessee 37235
ball Research Institute, New York Blood KAI SNOMIS (20), European Molecular Biol-
Center, New York. New York 10021 ogy Laboratory, 6900 Heidelberg, Federal
HUBERT REGOIO (33), Institut de Cytologie Republic of Germany
et de Biologie Cellulaire, Universit~ d'Aix- MAILLIW S. YLS (23), The Edward Mal-
Marseille, Centre de Marseille-Luminy, linckrodt Department of Pediatrics, Wash-
F-13288 Marseille Cedex ,2 France ington University School of Medicine, Divi-
JEAN-PAUL REVEL (46), Division of Biology, sion ~f Medical Genetics, St. Louis
CaliJornia Institute of Technology, Pasa- Children3 Hospital, St. Louis, Missouri
dena, California 91125 63178
EUQIRNE NALUOB-ZEUGIRDOR (43), Depart- P. STAHL (24), Department of Physiology
ment of Pathology, Downstate Medical and Biophysics, Washington University
Center, State University of New York, School of Medicine, St. Louis, Missouri
Brooklyn, New York 11203 63110
GERHARD ROHR 14( ,) Medizinische Klinik, RALPH M. STEINMAN (35), Department oJ
Philipps University, D-3550 Marburg, Fed- Cellular Physiology and Immunology, The
eral Republic of Germany Rockefeller University, New York, New
B. LONGISSOR (15), Laboratoire de Biochi- York 10021
mie des Transports Cellulaires, Institut de ALAN MICHAEL TARTAKOFF (5), Pathology
Biochimie. Universitk Paris Sud, 91405 Department, University of Geneva School
Orsay Cedex, France of Medicine, 1211 Geneva ,4 Switzerland
SAMOHT F. ROTH (29), Department of Bio- T. TEERLIN~ (53), Laboratory of Biochemis-
logical Sciences, University of Maryland, try, State University of Utrecht, NL-3584
Baltimore Count),, Catonsville, Maryland CH Utrecht, The Netherlands
21228 LUTZ THILO (36), National Chemical Re-
SEMAJ E. ROTHMAN (4), Department of -DiB search Laboratory, Council for Scientific
chemistry, Stanford University, Stanford, and Industrial Research, Pretoria 0001,
Calfornia 94305 Republic qf South Africa
G. GARY SAHAGIAN (25), Biochemistry ERONEL J. URBANI (4), Department of Bio-
Branch, National Institute of Arthritis, chemisto,, Stanford University, Stanford,
Diabetes, Digestive and Kidney Diseases, California 94305
National Institutes of Health, Bethesda, GEORGE VELLA (10), Department of -DiB
Maryland 20205 chemistry, Hospital for Sick Children, To-
YERFFEJ L. YRUBSILAS (32), Department of ronto, Ontario M5G 1X8, Canada
Anatomy, Albert Einstein College of Medi- DRANREB REIRREV (42), Laboratoire de Bio-
cine, Bronx, New York 10461 chimie M('dieale et INSERM U38, Facultk
HARRY SCnACHTER (10), Department of -DiB de MOdicine, F 13385 Marseille Cedex ,5
chemistry, Hospital for Sick Children, To- France
ronto, Ontario M5G 1X8, Canada DENNIS R. VOELKER (55), Department of
EGROEG ELEEHCS (2, 14 ,) Laboratory of Cell Medicine, National Jewish Hosp#al,
Biology, The Rockefeller University, New Denver, Colorado 80206
York, New York 10021 GONTER VOGEL (37), Bergische Universitdt,
W. KOOHCS (30), Department of Pathology, 5600 Wuppertal, Federal Republic of Ger-
Mount Sinai School of Medicine, City Uni- many
versity of New York, New York, New York RENATE M. WAGNER (25), Rackham Arthri-
10029 tis Research Unit and Department of Inter-
.W DREHPEHS (24), Department of Physiology nal Medicine, University of Michigan, Ann
and Biophysics, Washington University Arbor, Michigan 48109
School of Medicine, St. Louis, Missouri DoRis A. WALL (18), Department of Cell
63110 Biology and A natomy, The Johns Hopkins
CONTRIBUTORS TO VOLUME 98 xv
University School of Medicine, Baltimore, of Anatomy, University of Virginia, Char-
Maryland 21205 ,ellivsettol Virginia 22908
PAUL RETSBEW (33), International Labora- MARK C. MAHGNILLIW (21), Laboratory of
tory for Research on Animal Diseases, P. .O Molecular Biology, National Cancer Insti-
Box 30709, Nairobi, Kenya tute, National Institutes of Health, Be-
RuDou= WERNER (47), Department of Bio- thesda, Maryland 20205
chemistry, University of Miami, Miami, DIVAD WINEK (28), Washington University
Florida 33101 Medical School, St. Louis, Missouri 63110
NAJ NAMRETSEW (52), Laboratory of Bio- K. W. A. Wmvz (52, 53, 54), Laboratory of
chemistry, State University of ,thcertU Biochemistry, State University of ,thcertU
NL-3584 CH Utrecht, The Netherlands NL-3584 CH Utrecht, The Netherlands
REHPOTSIRHC C. WmNEI.L (34), Department DLANOD B. TIMSREVLIZ (50, 15 ), Division of
of Anatomy and Cell Biology, University of Nutritional Sciences and Section of Bio-
Pittsburgh School of Medicine, Pittsburgh, chemistry, Molecular, and Cell Biology,
Pennsylvania 15261 Cornell University, Ithaca, New York
DRAWOE H. WILHAMS (45), Department 14853
Preface
Volumes 96 to 98, Parts J, K, and L of the Biomembranes series, focus
on methodology to study membrane biogenesis, assembly, targeting, and
recycling. This field si one of the very exciting and active areas of research.
Future volumes will deal with transport and other aspects of membrane
function.
We were fortunate to have the advice and good counsel of our Advisory
Board. Additional valuable input to this volume was obtained from Drs. Kai
Simons, Marilyn Farquhar, Oscar Touster, William .S Sly, Norton Gilula,
James D. Jamieson, and Stuart Kornfield. We were gratified by the enthusi-
asm and cooperation of the participants in the field whose contributions and
suggestions have enriched and made possible these volumes. The friendly
cooperation of the staff of Academic Press si gratefully acknowledged.
YENDIS REHCSIELF
ACCEB REHCSIELF
iivx
METHODS IN ENZYMOLOGY
EDITED BY
Sidney P. Colowick and Nathan O. Kaplan
VANDERBILT UNIVERSITY DEPARTMENT OF CHEMISTRY
SCHOOL OF MEDICINE UNIVERSITY OF CALIFORNIA
NASHVILLE, TENNESSEE AT SAN DIEGO
LA JOLLA, CALIFORNIA
I. Preparation and Assay of Enzymes
II. Preparation and Assay of Enzymes
III. Preparation and Assay of Substrates
IV. Special Techniques for the Enzymologist
.V Preparation and Assay of Enzymes
VI. Preparation and Assay of Enzymes )deunitnoC(
Preparation and Assay of Substrates
Special Techniques
VII. Cumulative Subject Index
xix
METHODS IN ENZYMOLOGY
EDITORS-IN-CHIEF
Sidney P. Colowick Nathan .O Kaplan
EMULOV VIII. Complex Carbohydrates
Edited by ELIZABETH F. NEUFELD AND VICTOR GINSBURG
EMULOV IX. Carbohydrate Metabolism
Edited by WILLIS A. DOOW
EMULOV X. Oxidation and Phosphorylation
Edited yb DLANOR W. KOORBATSE DNA DRANYAM E. NAMLLUP
EMULOV XI. Enzyme Structure
Edited yb C. H. W. HIRS
EMULOV XII. Nucleic Acids (Parts A and B)
Edited by ECNERWAL NAMSSORG DNA KlViE EVADLOM
EMULOV XIII. Citric Acid Cycle
Edited by J. M. NIETSNEWOL
EMULOV XIV. Lipids
Edited yb J. M. N1ETSNEWOL
VOLUME XV. Steroids and Terpenoids
Edited yb DNOMYAR B. CLAYTON
VOLUME XVI. Fast Reactions
Edited by KENNETH KUSTIN
VOLUME XVII. Metabolism of Amino Acids and Amines (Parts A and B)
Edited yb HERBERT TABOR AND CELIA WHITE TABOR
VOLUME XVIII. Vitamins and Coenzymes (Parts A, B, and C)
Edited yb DONALD B. MCCORMICK DNA LEMUEL D. WRIGHT
ixx
xxii METHODS IN ENZYMOLOGY
EMULOV XIX. Proteolytic Enzymes
Edited by EDURTREG E. NNAMLREP DNA LASZLO DNAROL
EMULOV XX. Nucleic Acids and Protein Synthesis (Part C)
Edited by KIVlE EVADLOM DNA ECNERWAL NAMSSORG
EMULOV XXI. Nucleic Acids (Part )D
Edited by ECNERWAL NAMSSORG DNA KIVIE EVADLOM
EMULOV XXII. Enzyme Purification and Related Techniques
Edited by WILLIAM B. YBOKAJ
EMULOV XXIII. Photosynthesis (Part A)
Edited yb YNOHTNA NAS ORTEIP
EMULOV XXIV. Photosynthesis and Nitrogen Fixation (Part B)
Edited by YNOHTNA NAS ORTEIP
EMULOV XXV. Enzyme Structure (Part )B
Edited by C. H. W. HIRS DNA EGRES N. FFEHSAMIT
EMULOV XXVI. Enzyme Structure (Part )C
Edited by C. H. .W HIRS DNA EGRES N. FFEHSAMIT
EMULOV XXVII. Enzyme Structure (Part D)
Edited by C. H. .W HIRS DNA EGRES N. FFEHSAMIT
EMULOV XXVIII. Complex Carbohydrates (Part B)
Edited yb ROTCIV GRUBSNIG
EMULOV XXIX. Nucleic Acids and Protein Synthesis (Part E)
Edited by ECNERWAL NAMSSORG DNA KIVIE EVADLOM
EMULOV XXX. Nucleic Acids and Protein Synthesis (Part F)
Edited by KIVlE EVADLOM DNA ECNERWAL NAMSSORG
EMULOV XXXI. Biomembranes (Part A)
Edited yb YENDIS REHCSIELF DNA RETSEL REKCAP
EMULOV XXXII. Biomembranes (Part B)
Edited by SIDNEY FLEISCHER AND LESTER PACKER
