Methods in Enzymology Volume 171 Biomembranes Part R Transport Theory: Cells and Model Membranes EDITED BY Sidney Fleischer BeccaF leischer DEPARTMENT OF MOLECULAR BIOLOGY VANDERBILT UNIVERSITY NASHVILLE, TBNNESSEE Editorial Advisory Board Yaw0 Kagawa George Sachs Ronald Kaback Antonio Scarpa Martin Klingenberg Widmar Tanner Robert L. Post Karl Lnll-ich ACADEMIC PRESS, INC. Harcourt Brace Jovanovich, Publishers San Diego New York Berkeley Boston London Sydney Tokyo Toronto Contributors to Volume 171 elcitrA srebmun are in pmentheses following the names of contributors. snoitailiffA detsil are current. PER-AKE NOSSTREBLA (26), Department of V. QTOVSKY (41), Division of Molecular Biochemistry, Chemical Center, -revinU Plant Biology, University of ,ainrofilaC sity of Lund, S-221 0O Lund, Sweden Berkeley, Berkeley, ainrofilaC 02749 OLAF aPRAPS -~ NESREDNA (4), Department of CHRIS NESUALC (32), Department of -isyhP Physiology and Biophysics, Cornell Uni- ology and ,scisyhpoiB School of ,enicideM versity Medical ,egelloC New York, New Health Sciences ,retneC State ytisrevinU of York 12001 New York (SUNY) at Stony Brook, Stony RETEP .F R~,rAB I (40), Medical Research Brook, New York I 4971 Council (MRC) Secretory Mechanisms TREBRON A. DENCnER (13), Biophysics ,puorG Department of Physiology, Divi- ,puorG Department of ,scisyhP Freie -inU sion of Biomedical Sciences, King's Col- t~ltisrev Berlin, 0001-)1 Berlin ,33 Federal lege London, Strand, London WC2R 2LS, Republic of Germany England Rosa DEVS (5), Department of ygoloisyhP RETEP H. BARRY (35), School of ygoloisyhP and ,scisyhpoiB Faculty of Medicine, Uni- and ,ygolocamrahP The University of New versity of Chile, Casilla ,55007 ,ogaitnaS South Wales, Sydney, New South Wales, Chile Australia 2033 SIDNEY FLEISCHER (1), Department of Mo- R. BENZ (14), Lehrstuhlffir ,eigolonhcetoiB lecular Biology, Vanderbilt ,ytisrevinU t~itisrevinU ,grubzr~fW D-8700 ,grubzr~W ,ellivhsaN Tennessee 53273 Federal Republic of Germany J. KEvm FOSK~TT (39), Division of Cell Bi- DAVID CAFISO (16), Department of -simehC ,ygolo Research Institute, The Hospital for try, University of ,ainigriV ,ellivsettolrahC Sick Children, Toronto, Ontario, Canada ainigriV 10922 M5G 8X1 S. RoY CAPLAN (20), Department of Mem- ILAN GREBDEIRF (43), Department of -orciM brane Research, The Weizmann Institute ,ygoloib The George .S Wise Faculty of of ,ecneicS Rehovot ,00167 Israel Life Sciences, Tel-Aviv University, Tel- G. REUQSARRAC (31), Department of Medi- Aviv ,87996 Israel cine, University of Louisville, Louisville, PETER FROMHERZ (18), Abteilung f~r Bio- Kentucky 29204 physik, t~itisrevinU Ulm, 1)-7900 Ulm, N. YKSVONAJEHC (41), Laboratory of Mo- Donau ,7 Federal Republic of Germany ralucel and Cellular ,ygoloiB National In- E. FROMTER (33), Zentrum der ,eigoloisyhP stitute of Diabetes and Digestive and Kid- Johann Woifgang ,tirtisrevinU-ehteoG ney Diseases, National Institutes of 0006-)1 Frankfurt am Main ,07 Federal Health, Bethesda, Maryland 29802 Republic of Germany A. M. CHERET (21), Chargb de ,ehcrehceR ARLYN GARCIA-PEREZ (28), Laboratory of Institut National de al Sant~ et de al Re- Kidney and Electrolyte Metabolism, Na- ehcrehc M~dicale (INSERM) UIO, H@i. tional Heart, Lung and Blood Institute, tal Bichat, F-75877 Paris Cedex ,81 National Institutes of Health, Bethesda, ecnarF Maryland 29802 YRREJ D. GARDNER (29), Digestive Diseases I~d. ,hcnarB National Institute of Diabetes and ix X CONTRIBUTORS TO VOLUME 171 Digestive and Kidney Diseases, National Manchen, D-8000 M~nchen ,2 Federal Institutes of Health, Bethesda, Maryland Republic of ynamreG 29802 DEREK E. KNIOHT (40), Medical Research L. G. M. NODROG (33), Department of Phys- Council (MRC) Secretory Mechanisms iology, University of Otago, Medical ,puorG Department of ,ygoloisyhP Divi- ,loohcS Dunedin, New Zealand sion of Biomedical Sciences, King's -loC ACIDRON GREEN (37), Laboratory of Kidney lege London, Strand, London WC2R 2LS, and Electrolyte Metabolism, National England Heart, Lung and Blood Institute, National H.-A. KOLB (14), Department of ,ygoloiB Institutes of Health, Bethesda, Maryland ytisrevinU of Konstanz, D-7750 Konstanz 29802 ,1 Federal Republic of Germany TREBOR B. GUNN (8), Department of -isyhP OTIHAKAT ODNOK (10), First Department of ,ygolo Emory University School of Medi- Medicine, Hokkaido University School of ,enic Atlanta, aigroeG 22303 Medicine, Sapporo 060, Japan HPESOJ S. HANDLER (37), Laboratory of G. KOTTRA (33), Zentrum der ,eigoloisyhP Kidney and Electrolyte Metabolism, Na- Johann" Wolfgang ,tdtisrevinU-ehteoG tional Heart, Lung and Blood Institute, D.6000 Frankfurt am Main ,07 Federal National Institutes of Health, Bethesda, Republic of Germany Maryland 29802 DRAHNIER KRAMER (19), Institut f~r Bio- KURT OINNAH (25), Max-Planck-Institutfar eigolonhcet 1 der egalnasgnuhcsrofnreK ,eimehcoiB D-8033 Martinsried bei Man- Jalich GmbH, D-5170 Jalich, Federal Re- ,nehc Federal Republic of Germany public of Germany HANs-G. HCIRDIEH (25), Max-Planck-Insti- R. M. KRUPKA (5), Research Center, Agri- tut far Biochemie, D-8033 Martinsried bei erutluc ,adanaC London, ,oiratnO Canada Mfmchen, Federal Republic of Germany N6A 5B7 NOEL A. LEPPEH (43), Section of -simehcoiB PmLIP C. SIRAL (30), Department of -igoloiB try, Molecular and Cell Biology, Cornell cal Sciences, University of ,ainrofilaC ,ytisrevinU Ithaca, New York 35841 Santa ,arabraB Santa ,arabraB ainrofilaC PAUL KARL HORAN (27), Zynaxis Cell 60139 Science, Inc., Malvern, Pennsylvania P. LAUOER (14), Department of ,ygoloiB 55391 ytisrevinU of Konstanz, D-7750 Konstanz MAILLIW P. SKCNEJ (7), Graduate -trapeD ,1 Federal Republic of Germany ment of Biochemistry, Brandeis -revinU NHOJ P. REDAEL (38), Department of -isyhP sity, Waltham, Massachusetts 45220 ology, University of Otago, Medical ECURB D. JENSEN (27), Zynaxis Cell ,loohcS Dunedin, New Zealand Science, Inc., Malvern, Pennsylvania MI~UEL J. M. NXWEL (21), Unit~ de Re- 55391 cherches de ,eigolor~tnE-ortsaG Institut TREBOR T. NESNEJ (29), Digestive Diseases National de al Sant~ et de al Recherche ,hcnarB National Institute of Diabetes and M~dicale (INSERM) 1U ,O H~pital ,tahciB Digestive and Kidney Diseases, National F-75877 Paris Codex ,81 France Institutes of Health, Bethesda, Maryland NOMIS A. SIWEL (36), Department of -isyhP 29802 ygolo and ,scisyhpoiB University of Texas Rose M. ENOTSNHOJ (30), Department of Medical ,hcnarB ,notsevlaG Texas 05577 Biochemistry, McGill University, Mon- MAHARBA RETYOL (41 ), Department of -oiB treal ,cebeuQ Canada H3G 1 Y6 logical Chemistry, Institute of Life MARTIN GREBNEGNILK (2), Institut r~f ,secneicS The Hebrew University of Jeru- Physikalische Biochemie, Universitdt salem, Jerusalem ,40919 Israel CONTRIBUTORS TO VOLUME 171 xi KEvru D. LUSTIG (43), Departments of Bio- KEITH R. RUNYAN (8), Department of Medi- chemistry and Food Science and ,noitirtuN cine, Emory University School of Medi- ytisrevinU of ,aibmuloC-iruossiM Colum- ,enic Atlanta, aigroeG 22303 ,aib Missouri 11256 EOROEG SACHS (1), UCLA School of Medi- YNOHTNA D. C. MXCKNIGHT (38), -trapeD ,enic University of ,ainrofilaC Los ,selegnA ment of ,ygoloisyhP University of ,ogatO Los Angeles, ainrofilaC ,42009 and Center Medical ,loohcS Dunedin, New Zealand for Ulcer Research and Education ALAN McLAuOHLIN (16), Laboratory of (CURE), Veterans Administration, AV Metabolism, National Institute of lohoclA htrowsdaW Hospital ,retneC Los ,selegnA Abuse and Alcoholism, Rockville, Mary- ainrofilaC 37009 land 25802 MARTIN J. SANDERS (23), Department of TRAUTS McLAUGHLIN (16), Department of Metabolic Diseases and General Pharma- ygoloisyhP and ,scisyhpoiB School of Med- ,ygoloc Pfizer Central Research, ,notorG ,enici Health Sciences Center, State Uni- tucitcennoC 04360 versity of New York (SUNY) at Stony CARL YEFFEHCS (39), Department of -goloiB Brook, Stony Brook, New York 49711 ical Sciences, Columbia University, New R. B. McMICHENS (15), Laboratory of Mo- York, New York 72001 ralucel ,scisyhpoiB The University of Ala- GROEOSNAH SCHINDLER (11), Institute of bama at Birmingham, School of ,enicideM .scisyhpoiB University of Linz, A-4040 Birmingham, Alabama 49253 Linz/A ,fohu Austria RICHARD E. PAGANO (42), Department of M. ZTRAWHCS (31), Department of Medi- Embryology, Carnegie Institution of cine, University of Louisville, ,ellivsiuoL ,notgnihsaW Baltimore, Maryland 01212 Kentucky 29204 SOIRTEMED SOLUOPOIDAHAPAP (9), recnaC YNOHTNA W. OTTOCS (12), Division of Di- Research Institute, University of -rofilaC gestive ,sesaesiD Department of ,enicideM nia, San ,ocsicnarF San ,ocsicnarF Cali- Cornell University Medical ,egelloC New fornia 34149 York, New York 12001 OLE H. NESRETEP (34), Department of -syhP WILLIAM L. SMITH (28), Department of Bio- ,ygoloi University of ,loopreviL Liverpool chemistry, Michigan State University, L69 3BX, England East Lansing, Michigan 42884 DANmLA NOEORTEIP (20), CNR Unit for the ANDREW H. SOLL (23), UCLA School of Study of the Physiology of ,airdnohcotiM Medicine, University of ,ainrofilaC Los Institute of lareneG ,ygolohtaP University Angeles, Los Angeles, California ,42009 of ,audaP Padua, Italy and Center for reclU Research and -acudE ASEREHT P. WOLTERP (22), Institute of -aP tion (CURE), Veterans Administration, ,ygoloht Case Western Reserve ,ytisrevinU AV Wadsworth Hospital Center, Los An- School of Medicine, Cleveland, Ohio ,seleg ainrofilaC 37009 60144 WILLIAM K. GRUBNENNOS (28), Depart- SAMOHT G. WOLTERP (22), Institute of -aP ment of Biochemistry, Michigan State ,ygoloht Case Western Reserve ,ytisrevinU ,ytisrevinU East Lansing, Michigan 42884 School of Medicine, Cleveland, Ohio WILLIAM S. NAMLEIPS (28), Department of 60144 Physiology, Michigan State ,ytisrevinU WARREN S. REHM (31), Department of Med- East Lansing, Michigan 42884 icine, University of Louisville, ,ellivsiuoL G. STARK (14), Department of ,ygoloiB Uni- Kentucky 29204 versity of Konstanz, D-7750 Konstanz ,1 HAGAI GREBNETTOR (17), Pathology De- Federal Republic of Germany partment, Hahnemann University, Phila- KCIREDOR E. STEELE (37), Laboratory of ,aihpled Pennsylvania 20191 Technical Development, National Heart, xii CONTRIBUTORS TO VOLUME 171 Lung and Blood Institute, National Insti- C. M. MALAHCATAKNEV (15), Polygen -roC tutes of Health, Bethesda, Maryland ,noitarop Waltham, Massachusetts 45120 29802 GARY A. WEISMAN (43), Departments of DERFLIW D. STEIN (3), Department of Bio- Biochemistry and Food Science and Nutri- logical Chemistry, Institute of Life tion, University of ,aibmuloC-iruossiM ,secneicS The Hebrew University of -ureJ ,aibmuloC Missouri 11256 salem, Jerusalem ,40919 Israel JOHN WELLS (24), Department of Internal Medicine, Pulmonary Division, University T. L. ENAPART (15), Laboratory of -uceloM of Texas Medical Branch, ,notsevlaG lar ,scisyhpoiB The University of Alabama Texas 05577 at Birmingham, School of Medicine, Bir- mingham, Alabama 49253 YCNAN K. WILLS (36), Department of -isyhP ygolo and ,scisyhpoiB University of Texas DAN W. URRY (15), Laboratory of -uceloM Medical ,hcnarB ,notsevlaG Texas 05577 ral ,scisyhpoiB The University of Alabama YNOHTNA WINxsra (16), Department of -oiB at Birmingham, School of Medicine, Bir- chemistry, State University of New York mingham, Alabama 49253 (SUNY) at Stony Brook, Stony Brook, New PAUL S. USTER (42), Liposome ,ygolonhceT York 49711 Inc., Menlo Park, ainrofilaC 52049 MARTIN C. ELDOOW (9), Liposome Tech- TTUDYAJ V. VADGAMA (6), Division of nology, Inc., Menlo Park, California Nephrology and Hypertension, Depart- 52049 ment of Medicine, LAC Harbor-UCLA DAVID ZAKIM (12), Division of Digestive Medical Center, UCLA School of Medi- ,sesaesiD Department of Medicine, Cornell ,enic University of ,ainrofilaC Los ,selegnA University Medical College, New York, ,ecnarroT ainrofilaC 90509 New York 12001 Preface Biological transport is part of the Biomembranes series of Methods in Enzymology. It is a continuation of methodology concerned with mem- brane function. This is a particularly good time to cover the topic of biological membrane transport because there is now a strong conceptual basis for its understanding. The field of transport has been subdivided into five topics. .1 Transport in Bacteria, Mitochondria, and Chloroplasts 2. ATP-Driven Pumps and Related Transport .3 General Methodology of Cellular and SubceUular Transport 4. Cellular and Subeellular Transport: Eukaryotic (Nonepithelial) Cells 5. Cellular and Subcellular Transport: Epithelial Cells Topic 1 covered in Volumes 521 and 621 initiated the series. Topic 2 is covered in Volumes 651 and 157. Topic 3 is covered in Volumes 171 and 172. The remaining two topics will appear in subsequent volumes of the Biomembranes series. Topic 3 provides general methodology and concepts. Its coverage in- cludes transport theory and general methodology for the study ofbiomem- branes. It is divided into two parts: this volume (Part R) which covers Transport Theory: Cells and Model Membranes and Volume 172 (Part S) which covers Transport: Membrane Isolation and Characterization. We are fortunate to have the good counsel of our Advisory Board. Their input ensures the quality of these volumes. The same Advisory Board has served for the complete transport series. Valuable input on the outlines of the five topics was also provided by Qais A1-Awqati, Ernesto Carafoli, Halvor Christensen, Isadore Edelman, Joseph Hoffman, Phil Kuauf, and Hermann Passow. The names of our advisory board members were inadvertanfly omitted in Volumes 521 and 126. When we noted the omission, it was too late to rectify the problem. For Volumes 521 and 126, we are pleased to acknowl- edge the advice of Angelo Azzi, Youssef Hatefi, Dieter Oesterhelt, and Peter Pedersen. The enthusiasm and cooperation of the participants have enriched and made these volumes possible. The friendly cooperation of the staff of Academic Press is gratefully acknowledged. iiix xiv ECAFERP These volumes are dedicated to Professor Sidney Colowick, a dear friend and colleague, who died in 1985. We shah miss his wise counsel, encouragement, and friendship. YENDIS REHCSIELF ACCEB REHCSIELF METHODS IN ENZYMOLOGY EMULOV I. Preparation and Assay of Enzymes Edited by YENDIS P. KCIWOLOC DNA NAHTAN O. NALPAK EMULOV II. Preparation and Assay of Enzymes Edited by YENDIS P. KCIWOLOC DNA NAHTAN O. NALPAK EMULOV III. Preparation and Assay of Substrates Edited by YENDIS P. KCIWOLOC DNA NAHTAN O. NALPAK EMULOV IV. Special Techniques for the Enzymologist Edited by YENDIS P. KCIWOLOC DNA NAHTAN O. NALPAK EMULOV V. Preparation and Assay of Enzymes Edited by YENDIS P. KCIWOLOC DNA NAHTAN O. NALPAK EMULOV VI. Preparation and Assay of Enzymes )deunitnoC( Preparation and Assay of Substrates Special Techniques Edited by YENDIS P. KCIWOLOC DNA NAHTAN O. NALPAK EMULOV VII. Cumulative Subject Index Edited by YENDIS P. KCIWOLOC DNA NAHTAN O. NALPAK EMULOV VIII. xelpmoC Carbohydrates Edited by HTEBAZILE F. DLEFUEN DNA ROTCIV GRUBSNIG EMULOV IX. Carbohydrate Metabolism Edited by SILLIW A. DOOW EMULOV X. Oxidation and Phosphorylafion Edited by DLANOR W. KOORBATSE DNA DRANYAM E. NAMLLUP EMULOV XI. Enzyme Structure Edited by C. H. W. SRIH EMULOV XII. Nucleic Acids (Parts A and B) Edited by ECNERWAL NAMSSORG DNA EIVIK EVADLOM XV xvi METHODS IN ENZYMOLOGY EMULOV XIII. Citric Acid Cycle Edited by J. M. NIETSNEWOL EMULOV XIV. Lipids Edited by J. M. NIETSNEWOL EMULOV XV. Steroids and Terpenoids Edited by DNOMYAR .B NOTYALC EMULOV XVI. Fast Reactions Edited by HTENNEK NITSUK EMULOV XVII. Metabolism of Amino Acids and Amines (Parts A and )B Edited by TREBREH ROBAT DNA AILEC ETIHW ROBAT EMULOV XVIII. Vitamins and Coenzymes (Parts A, ,B and C) Edited by DLANOD .B KCIMROCCM DNA LEUMEL D. THGIRW EMULOV XIX. Proteolytic Enzymes Edited by EDURTREG E. NNAMLREP DNA OLZSAL DNAROL EMULOV XX. Nucleic Acids and Protein Synthesis (Part C) Edited by EIVIK EVADLOM DNA ECNERWAL NAMSSORG EMULOV XXI. Nucleic Acids (Part D) Edited by ECNERWAL NAMSSORG DNA EIVIK EVADLOM EMULOV XXII. Enzyme Purification and Related Techniques Edited by MAILLIW .B YBOKAJ EMULOV XXIII. Photosynthesis (Part A) Edited by 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. SRIH DNA EGRES N. FFEHSAMIT EMULOV XXVI. Enzyme Structure (Part C) Edited by C. H. W. SRIH DNA SERGE N. FFEHSAMIT METHODS IN ENZYMOLOGY xvii EMULOV XXVII. Enzyme Structure (Part D) Edited by C. H. W. SRIH DNA EGRES N. FFEHSAMIT EMULOV XXVIII. Complex Carbohydrates (Part )B Edited by ROTCIV GRUBSNIG EMULOV XXIX. Nucleic Acids and Protein Synthesis (Part E) Edited by ECNERWAL NAMSSORG DNA EIVIK EVADLOM EMULOV XXX. Nucleic Acids and Protein Synthesis (Part F) Edited by EIVIK EVADLOM DNA ECNERWAL NAMSSORG EMULOV XXXI. Biomembranes (Part A) Edited by YENDIS REHCSIELF DNA RETSEL REKCAP EMULOV XXXII. Biomembranes (Part )B Edited by YENDIS REHCSIELF DNA RETSEL REKCAP EMULOV XXXIII. Cumulative Subject Index Volumes I-XXX Edited by AHTRAM G. SINNED DNA DRAWDE A. SINNED EMULOV XXXIV. Affinity Techniques (Enzyme Purification: Part )B Edited by MAILLIW .B YBOKAJ DNA MEIR KEHCLIW EMULOV XXXV. tipid$ (Part )B Edited by NHOJ M. NIETSNEWOL EMULOV XXXVI. Hormone Action (Part A: Steroid Hormones) Edited by TREB W. YELLAM'O DNA LEOJ G. NAMDRAH EMULOV XXXVII. Hormone Action (Part :B Peptide Hormones) Edited by TREB W. YELLAM'O DNA LEOJ G. NAMDRAH EMULOV XXXVIII. Hormone Action (Part :C Cyclic Nucleotides) Edited by LEOJ G. NAMDRAH DNA TREB W. YELLAM'O EMULOV XXXIX. Hormone Action (Part D: Isolated Cells, Tissues, and Organ Systems) Edited by LEOJ G. NAMDRAH DNA TREB W. YELLAM'O EMULOV XL. Hormone Action (Part :E Nuclear Structure and Function) Edited by TREB W. YELLAM'O DNA LEOJ G. NAMDRAH