Table Of ContentAdvisory Board
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Keith R. Porter (Chuirmun)
Elizabeth D. Hay George E. Palade
T. C. Hsu George D. Pappas
Dan Mazia Jean-Paul Revel
David Sabatini
METHODS IN CELL
BIOLOGY
Preppcired under the Auspices of the American Society for Cell Biology
VOLUME 23
Basic Mechanisms of Cellular Secretion
Edited by
ARTHUR R. HAND AND CONSTANCE OLIVER
LABORATORY OF BIOLOGICAL STRUCTURE
NATIONAL INSTITUTE OF DENTAL RESEARCH
NATIONAL INSTITUTES OF HEALTH
BETHESDA. MARYLAND
1981
ACADEMIC PRESS
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LIST OF CONTRIBUTORS
Numbers in pcirentheses ittdicute the puges on which the authors' conrributions begirt,
D. BANERJEETh, e Lindsley F. Kimball Re- ANNA M. CASTLES, ection of Cell Biology,
search Institute of the New York Blood Yale University School of Medicine, New
Center. 310 East 67th Street, New York, New Haven, Connecticut 06510 (335)
York 10021 (231)
J. DAVIDCA STLES,e ction of Cell Biology, Yale
PHILIPJ. BASSFORDJ, R.,' Department of Mic- University School of Medicine, New Haven,
robiology and Molecular Genetics, Harvard Connecticut 06510 (335)
Medical School, Boston, Massachusetts
02115 (27) CONSTANCP.E C HRISTIADNe,p artment of Oral
Diagnosis, University of Connecticut School
JON BECKWITHD,e partment of Microbiology of Dental Medicine, Farmington, Connecticut
and Molecular Genetics, Harvard Medical 06032 (531)
School, Boston, Massachusetts 021 15 (27)
JEAN-MARICEL BMENTU, nite de Programma-
HUGHESB EWUELLEU,n ite de Progranimation tion Moleculaire et Toxicologie Genetique,
Moleculaire et Toxicologie Genetique, lnstitut lnstitut Pasteur, 75015 Paris, France (27)
Pasteur, 75015 Paris, France (27)
Y. CLERMONDTe,p artment of Anatomy, McGill
RICHARDA . BERG,^ Pulmonary Branch, Na- University, Montreal, Quebec, Canada H3A
tional Heart, Lung and Blood Institute, Na- 282 (155)
tional Institutes of Health, Bethesda, Mary-
land 20205 (I 19) CARLE . CREUTZS,e ction on Cell Biology and
Biochemistry of the Clinical Hematology
N. BORGESED,e partment of Pharmacology and Branch, National Institute of Arthritis.
CNR Center of Cytopharmacology, Univer- Metabolism and Digestive Diseases, National
sity of Milan, 20129 Milan, Italy (445) Institutes of Health, Bethesda. Maryland
20205 (313)
BRIANBO TTAROD,e partment of Oral Diagnosis,
University of Connecticut School of Dental LESLIES . CUrI.ER, Department of Oral Diag-
Medicine, Farmington, Connecticut 06032 nosis, University of Connecticut School of
(53 I) Dental Medicine, Farmington, Connecticut
06032 (531)
MARCIABU DARF,' Department of Chemistry,
University of Oregon, Eugene, Oregon 97403 JEFFREY M. DAVIDSONPu,~lm onary Branch,
(101) National Heart, Lung and Blood Institute, Na-
'Present uddress: Department of Bacteriology and Immunology, University of North Carolina
School of Medicine, Chapel Hill, North Carolina 27514.
2Present uddress: Department of Biochemistry, College of Medicine and Dentistry of New Jersey,
Rutgers University Medical School, Piscataway. New Jersey 08854.
'Present uddress: Department of Molecular Biology, University of California, Berkeley, Califor-
nia 94720.
4Presenr ciddress: Department of Pathology, College of Medicine, University of Utah, Salt Lake
City, Utah 84132.
xv
xvi LIST OF CONTRIBUTORS
tional Institutes of Health, Bethesda, Mary- sachusetts General Hospital, and Howard
land 20205 (I 19) Hughes Medical Institute Laboratories, Har-
vard Medical School, Boston, Massachusetts
J. S. DAVISONT,h e Physiological Laboratory, 02114 (51)
University of Liverpool, Liverpool L69 3BX,
United Kingdom (513) ARTHURR . HAND,L aboratory of Biological
Structure, National Institute of Dental Re-
W. W. DOUGLADS,e partment of Pharmacology, search, National Institutes of Health,
Yale University School of Medicine, New Bethesda, Maryland 20205 (I, 137, 429)
Haven, Connecticut 065 10 (483)
JOE HEDGPETHU,~n ite de Programmation
SCOTTD . EMR? Department of Microbiology Moleculaire et Toxicologie Genetique, Institut
and Molecular Genetics, Harvard Medical Pasteur, 75015 Paris, France (27)
School, Boston, Massachusetts 021 15 (27)
EDWARDHE RBERTD, epartment of Chemistry,
MARILYNGI ST FARQUHARSe,c tion of Cell University of Oregon, Eugene, Oregon 97403
Biology, Yale University School of Medicine,
(101)
New Haven, Connecticut 06510 (399)
L. HERMO,D epartment of Anatomy, McGill
H. DAVIDF ISCHERD, epartments of Pediatrics
University, Montreal, Quebec, Canada H3A
and Genetics, Washington University School
282 (155)
of Medicine, and Division of Medical Gene-
tics, St. Louis Children’s Hospital, St. Louis,
SYLVITA. HOFFSTEIDNe, partment of Medicine,
Missouri 631 10 (191)
Division of Rheumatology, New York Uni-
versity School of Medicine, New York, New
D. V. GALLACHETRh,e Physiological Labora-
York 10016 (259)
tory, University of Liverpool, Liverpool L69
3BX, United Kingdom (513)
MAURICE HOFNUNGu,n ite de Programmation
Moleculaire et Toxicologie Genetique, Institut
ALFONSGOO NZALEZ-NORIEDGepAar,t ments of
Pasteur, 75015 Paris, France (27)
Pediatrics and Genetics, Washington Univer-
sity School of Medicine, and Division of Med-
ical Genetics, St. Louis Children’s Hospital, ERICH OLTZMANDe, partment of Biological Sci-
St. Louis, Missouri 631 10 (191) ences, Columbia University, New York, New
York 10027 (379)
JEFFREYH . GRUBBD, epartments of Pediatrics
and Genetics, Washington University School WAYNLE. HUBBELDL,e partment of Chemistry,
of Medicine, and Division of Medical Gene- University of California, Berkeley, California
tics, St. Louis Children’s Hospital, St. Louis, 94720 (335)
Missouri 631 10 (191)
N. IWATSUKI,’ The Physiology Laboratory,
JOEL F. HABENELRa,b oratory of Molecular En- University of Liverpool, Liverpool L69 3BX,
docrinology and Endocrine Unit, Mas- United Kingdom (513)
SPresen~ad dress: Cancer Biology Program, NCI Frederick Cancer Research Center, Frederick,
Maryland 21701.
6Present address: Howard Hughes Medical Institute, Department of Biochemistry, University of
California, San Francisco, California 94143.
’Presenf address: Department of Applied Physiology, Tohoku University School of Medicine,
Seiryocho 2-1, Sendai, Japan 980.
LIST OF CONTRIBUTORS xvii
JAMES D. JAMIESONS,e ction of Cell Biology. tics, St. Louis Children's Hospital, St. Louis,
Yale University School of Medicine, New Missouri 63110 (191)
Haven, Connecticut 065 10 (547)
CONSTANCOE LIVERL, aboratory of Biological
Structure, National Institute of Dental Re-
HELENM . KORCHAKD,e partment of Medicine,
search, National Institutes of Health,
Division of Rheumatology, New York Uni-
versity School of Medicine, New York, New Bethesda, Maryland 20205 (I, 137, 429)
York 10016 (461)
LELIOO RCI, Institute of Histology and Em-
bryology, University of Geneva Medical
HENRY M. KRONENBERCL, aboratory ot
School, 1211 Geneva 4, Switzerland (51,
Molecular Endocrinology and Endocrine
283)
Unit, Massachusetts General Hospital, and
Howard Hughes Medical Institute RICHARIL). ORNBERCS. ection on Functional
Laboratories, Harvard Medical School, Bos- Neuroanatomy, Laboratory of Neuropathol-
ton, Massachusetts 021 14 (51) ogy and Neuroanatomical Sciences, National
Institutes of Health, Bethesda, Maryland
R. LAUCIERT,~he Physiological Laboratory, 20205, and Marine Biological Laboratory,
University of Liverpool, Liverpool L69 3BX, Woods Hole, Massachusetts 02543 (301)
United Kingdom (513)
CHRISTOPPHA TZELTD, epartment of Biochemis-
try, University of Chicago, Chicago, Illinois
C. P. LEBLOND,D epartment of Anatomy,
McGill University, Montreal, Quebec. 60637 (73)
Canada H3A 2B2 (167)
CHRisroPHER J. PAZOLES,~S"e ction on Cell
Biology and Biochemistry of the Clinical
HARVEYF . Looisti, Department of Biology.
Hematology Branch, National Institute of Ar-
Massachusetts Institute of Technology, Cam-
thritis, Metabolism and Digestive Diseases,
bridge, Massachusetts 02 I39 (5)
National Institutes of Health, Bethesda, Mary-
land 20205 (313)
MARIA-LUISAM ACCECCHINI,' Department at
Biochemistry, Biocenter, University of Basel, G. T. PEARSONT,h e Physiological Laboratory,
Ch-4056 Basel, Switzerland (39) University of Liverpool, Liverpool L69 3BX,
United Kingdom (513)
1. MELWLESI,D epartment of Pharmacology
A. PERRELETI,n stitute of Histology and Em-
and CNR Center of Cytopharmacology, Uni-
bryology, University of Geneva Medical
versity of Milan, 20129 Milan, Italy (445)
School, 121 I Geneva 4, Switzerland (283)
R. MONTESANOIn, stitute of Histology and Em- 0. H. PETERSENT,h e Physiological Laboratory,
bryology, University of Geneva Medical
University of Liverpool, Liverpool L69 3BX,
School, 121 I Geneva 4, Switzerland (283) United Kingdom (513)
MARVINNA TOWICZD. epartments of Pediatrics MARJORIEPb iii.LiPs,ii Department of Chemis-
and Genetics, Washington University School try, University of Oregon, Eugene, Oregon
of Medicine, and Division of Medical Gene- 97403 (101)
*Preserir address: INSERM Unite 31, 46 Chemin de la Gaye, 13009 Marseille, France.
'Prrserir address: International Minerals & Chemical Corporation, P.O. Box 207, Terre Haute,
Indiana 47808.
'"Presen/ ciddress: Pfizer Central Research, Eastern Point Road, Groton, Connecticut 06340.
"Presenr address; Laboratory of Toxicology, Harvard School of Public Health, Boston, Mas-
sachusetts 021 15.
xviii LIST OF CONTRIBUTORS
H. G. Ptiii..mii, The Physiological Laboratory. sachusetts Institute of Technology, Cam-
University of Liverpool, Liverpool L69 3BX, bridge, Massachusetts 02139 (89)
United Kingdom (5 13)
NAPHTALI SAVION,T”h e Department of Biolog-
HARVEYB. POI.I.ARDS, ection on Cell Biology ical Chemistry, The Hebrew University of
and Biochemistry of the Clinical Hematology Jerusalem, Jerusalem, Israel (359)
Branch, National Institute of Arthritis,
Metabolism and Digestive Diseases, National GEORGEA . SCHEELEC,e ll Biology Department,
Institutes of Health, Bethesda, Maryland The Rockefeller University, New York, New
20205 (313) York 10021 (345)
MARYP ORTER,D epartment of Biology, Mas-
ZVIS ELINGERT,h e Department of Biological
sachusetts Institute of Technology, Cam-
Chemistry, The Hebrew University of
bridge, Massachusetts 02 I39 (5)
Jerusalem, Jerusalem, Israel (359)
JOHN T. POT.IS, JR., Laboratory of Molecular
Endocrinology and Endocrine Unit, Mas- THOMASJ . SILHAVY,D~e~pa rtment of Mi-
sachusetts General Hospital, and Howard crobiology and Molecular Genetics. Harvard
Medical School, Boston, Massachusetts
Hughes Medical lnstitute Laboratories, Har-
021 15 (27)
vard Medical School, Boston, Massachusetts
021 I4 (51)
WILLIASM. SLY,D epartments of Pediatrics and
JAMESW . PUTNEYJ, R.,D~e~pa rtment of Phar- Genetics, Washington University School of
macology, Wayne State University, School of Medicine, and Division of Medical Genetics,
Medicine, Detroit, Michigan 48201 (503) St. Louis Children’s Hospital, St. Louis, Mis-
souri 631 10 (191)
A. RAMBOURGD,e partement de Biologie du
Commissariat I’Energie Atomique, 91 190 JAMESE . SMOLEND, epartment of Medicine,
Saclay, France (155) Division of Rheumatology, New York Uni-
versity School of Medicine, New York, New
C. M. REDMANT, he Lindsley F. Kimball Re-
York 10016 (461)
search Institute of the New York Blood
Center, 310 East 67th Street, New York, New
MARTIND . SNIDERD,e partment of Biology and
York 10021 (231)
Center for Cancer Research, Massachusetts
Institute of Technology, Cambridge, Mas-
THOMASS . REESE, Section on Functional
sachusetts 02 I39 (89)
Neuroanatomy , Laboratory of Neuropathol-
ogy and Neuroanatomical Sciences, National
DONALDF. STEINERD,e partment of Biochemis-
Institutes of Health, Bethesda, Maryland
20205, and Marine Biological Laboratory, try, University of Chicago, Chicago, Illinois
Woods Hole, Massachusetts 02543 (301) 60637 (73)
PHILLIPWS . ROBBINSD, epartment of Biology THOMASP . STOSSEL,H ematology-Oncology
and Center for Cancer Research, Mas- Unit, Massachusetts General Hospital, and
ILPresent uddress: Department of Pharmacology, Medical College of Virginia, Virginia Com-
monwealth University, Richmond, Virginia 23298.
”Present uddress: The Lautenberg Center for General and Tumor Immunology, Hadassah Medi-
cal School, The Hebrew University of Jerusalem, Jerusalem, Israel.
I4Present uddress: Cancer Biology Program, NCI Frederick Cancer Research Center, Frederick,
Maryland 21701.
LIST OF CONTRIBUTORS xix
Department of Medicine, Harvard Medical JOHN A. WILLIAMDS,e partment of Physiology,
School, Boston, Massachusetts 021 14 (215) University of California. San Francisco,
California 94143 (247)
HOWARDS. TACERD, epartment of Biochemia-
GLENDAM . WRIGHT."D epartment of Anatomy,
try, University of Chicago, Chicago, Illinois
McGill University, Montreal, Quebec, Canada
60637 (73)
H3A 2B2 (167)
STUARJT. WEISS.D epartment of Pharmacology.
S. Yu. The Lindsley F. Kimball Research Insti-
Wayne State University. School of Medicine,
tute of the New York Blood Center, 310 East
Detroit, Michigan 48201 (503)
67th Street, New York, New York 10021
(231)
GERAL.WD EISSMANNDe, partment of Medicine,
Division of Rheumatology, New York Uni- ASHERZ ILBERSTEIN, Department Of Biology,
versity School of Medicine, New York, New Massachusetts Institute of Technology, Cam-
York, 10016 (461) bridge, Massachusetts 02139 (5)
"Presrn/ ut/dres.s: Department of Zoology, University of Toronto, Toronto, Ontario, Canada M5S
IAl.
PREFACE
Basic Mechanisms of Cellular Secretion had its beginnings in the Conference
sponsored by the National Institute of Dental Research on Basic Mechanisms of
Cellular Secretion, held in Annapolis, Maryland, September 17-21, 1979. In
planning this conference, it became apparent that there was no publication which
addressed itself solely to the various aspects of the secretory process. The study
of the secretory process crosses interdisciplinary lines and utilizes all the tech-
niques available to the cell biologist. This book integrates the various methods as
they are used to investigate specific aspects of secretion. Each of the six sections
of the book deals with a major area of research related to the secretory process.
The introductory chapter in each section provides a framework for several shorter
chapters dealing with specific experimental systems. The concluding chapter
synthesizes the contents of the book into a unified view of secretion, and dis-
cusses several areas which warrant further exploration. It is hoped that Basic
Mechanisms of Cellular Secretion will serve as a source book both for estab-
lished investigators wishing to broaden the scope of their research efforts and for
individuals just beginning to delve into the complexities of the secretory process.
Basic Mechanisms of Cellular Secretion is the result of the combined efforts of
many individuals. We are especially grateful to Drs. David B. Scott, Director,
NIDR, and Marie U. Nylen, Director of Intramural Research, NIDR, for their
continued support and encouragement. We would also like to thank the authors
for their contributions, and Mrs. Patricia Youmans for her editorial and adminis-
trative assistance.
ARTHURR . HAND
CONSTANCOEL IVER
xxi
METHODS IN CELL BIOLOGY, VOI.UME 23
Chapter 1
In troda cti on
A. R. HAND ANDC. OLIVER
Laboratory of Biological Structures,
National Institute of Dental Research,
hiational Institirtes of Health,
Bethesda. Maryland
Text . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
The secretion of proteins is a functional characteristic of virtually all cells.
Single-cell organisms respond to and interact with their environment by the
secretion of proteins. The cells of higher organisms secrete proteins to create or
alter their environment, to influence the activity of other cells in their immediate
vicinity or at some distance, or to digest food substances. Thus, it is not surpris-
ing that the processes involved in the synthesis and secretion of proteins have
been so intensively and extensively studied during the last three decades. The
basic steps in protein secretion, as outlined by Palade (1975), are (I) synthesis;
(2) segregation; (3) intracellular transport; (4) concentration; (5) intracellular
storage; and (6) discharge. The generality of these steps has been confirmed by
studies of numerous cells and tissues, and as the title of this volume implies, the
basic mechanisms underlying these steps are remarkably similar for organisms as
diverse as bacteria and mammals. While the prototype secretory cell, the pan-
creatic exocrine cell, continues to be widely used, fibroblasts, leukocytes,
plasma cells, virus-infected cells, prokaryotes, and isolated organelles have been
established as important models for studying various aspects of protein secretion.
Thus, much of our knowledge of the secretory process has been gleaned from
systems which express their individuality through exaggeration, or deletion, of
one or more of the steps.
I
a
Copyright 1Y81 by Academic Pres. Inc
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