Membrane Transport in Biology - Volume I

Membrane Transport in Biology

Edited by

G. Giebisch . D. C. Tosteson H. H. Ussing

Associate Editor

M. T. Tosteson

Volume I

Concepts and Models

Contributors

0. S. Andersen J. E. Hall D. J. Hanahan U. V. Lassen P. K. Lauf R. J. Lefkowitz E. Racker B. E. Rasmussen

S. A. Rudolph F. A. Sauer C. W. Slayman G. Stark O. Sten-Knudsen H. H. Ussing

Editor

D. C. Tosteson

With 108 Figures and 36 Tables

Springer-Verlag Berlin' Heidelberg' New York 1978

Professor Dr. Gerhard Giebisch Yale University, School of Medicine, Department of Physiology

333 Cedar Street, New Haven, Conn. 06510/ USA

Professor Dr. Daniel C. Tosteson, Dean Harvard Medical School

25 Shattuck Street, Boston, Mass. 02115 / USA

Professor Dr. Hans H. Ussing University of Copenhagen, Institute of Biological Chemistry A

13 Universitetsparken, DK - 2100 Copenhagen

Dr. Magdalena T. Tosteson Harvard Medical School, Department of Physiology

25 Shattuck Street, Boston, Mass. 02115 / USA

ISBN-13: 978-3-642-46372-3 DOl: 10.1007/978-3-642-46370-9

e-ISBN-13: 978-3-642-46370-9

Library of Congress Cataloging in Publication Data: Main entry under title: Membrane transport in biology. 1. Biological transport. 2. Membranes (Biology). I. Giebisch, G., 1927-; II. Tosteson, D. C, 1925-; III. Ussing, Hans H., 1911-. [DNLM: 1. Biological transport. 2. Cell membrane -

Physiology. QH 509 M 533] QH 509-M44. 574.8'75. 78-17669.

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Preface

This Volume forms the cornerstone of this series of four books on Membrane Transport in Biology. It includes chapters that address i) the theoretical basis of investigations of transport processes across biological membranes, ii) some of the experimental operations often used by scientists in this field, iii) chemical and biological properties common to most biological membranes, and iv) planar thin lipid bilayers as models for biological membranes. The themes developed in these chapters recur frequently throughout the entire series.

Transport of molecules across biological membranes is a special case of diffu­sion and convection in liquids. The conceptual frame of reference used by investigators in this field derives, in large part, from theories of such processes in homogeneous phases. Examples of the application of such theories to transport across biological membranes are found in Chapters 2 and 4 of this Volume. In Chapter 2, Sten-Knudsen emphasizes a statistical and molecular approach while, in Chapter 4 Sauer makes heavy use of the thermodynamics of irreversi­ble processes. Taken together, these contributions introduce the reader to the two sets of ideas which have dominated the thinking of scientists working in this field. Theoretical consideration of a more special character are also included in several other Chapters in Volume I. For example, Ussing (Chapter 3) re-works the flux ratio equation which he introduced into the field of transport across biological membranes in 1949. Andersen (Chapter 11) discusses some of the physico-chemical properties of bilayers which place constraints on the applica­bility of the theory of transport processes in homogeneous systems to these extrem­ely thin and highly organized structures. Stark (Chapter 12) treats quite thor­oughly the theory of carrier mediated ion transport across bilayers. These essays describe many of the concepts now available to and used by investigators of membrane transport in biology.

Measurements of the rates of transport of substances across membranes in­volve certain common experimental operations. In Chapter 3, Ussing reviews conceptual and technical aspects of isotopic tracers as non-perturbing probes of transport processes. He was one of the first scientists to use these techniques when radioactive isotopes became available in the 1940's and has continued to observe and participate in the gathering of information with these tools since that time. Through the use of isotopic tracers, the operational description of net transport processes as the resultant of oppositely directed unidirectional fluxes

VI Preface

became possible for the first time, thus deepening our grasp of the kinetic mechanisms underlying these phenomena. For example, measurements of fluxes with tracers permitted Ussing to recognize exchange diffusion as an important and hitherto unsuspected pathway for the mixing of molecules separated by membranes. Electrical methods have also been usefully employed to character­ize the transport of ions across membranes. Lassen and Rasmussen review the advantages and pitfalls of these techniques when applied to cell membranes in Chapter 5. They point out that the introduction of microelectrodes into small cells may seriously perturb the system under investigation. Several applications of electrical methods to detect ion movements within and across membranes are included in the three last chapters on bilayers.

Biological membranes share certain molecular properties which influence all transport processes. These shared characteristics form the substance of several papers in this Volume. Hanahan introduces these with a thoughtful review of the chemical composition of red cell membranes. Racker contributes a brilliant analysis of membrane enzymes with special emphasis on ATPases (Chapter 8). Slayman reviews the genetic determination of membrane transport systems in Chapter 7. Lauf explores the relation between immunological reactions and transport phenomena. Rudolph and Lefkowitz provide an introduction to the rapidly growing contemporary literature on membrane receptors which modu­late and regulate transport. These Chapters are both distinctive expressions of important dimensions of membrane transport in biology and also useful summa­ries of material which will be helpful to readers of subsequent Volumes in this series.

The ingenious work of Mueller and Rudin in the early 60's began a new stage of research on transport across biological membranes. Before that time, the minds of most scientists in the field were animated by concepts derived from the analysis of bulk systems of membranes with dimension and physical properties that are markedly different from the structures which surround cells and organ­elles. Their development of relatively stable thin lipid bilayers made available for the first time a model with chemical composition and thickness comparable to the membranes found in living cells. The planar character of Mueller-Rudin membranes permitted simultaneous electrical and chemical transport measure­ments. The large number of productive insights into membrane transport which have resulted from studies of these structures are summarized in the last three chapters of this Volume. Andersen describes the features of unmodified bilay­ers, Stark considers carriers and Hall discusses channels. The relatively simple and defined character of these systems permits quantitative analysis of many concepts which are valuable in thinking about transport across more complicat­ed biological membranes

New Haven, Boston, Copenhagen G. Giebisch D. C. Tosteson H. H. Ussing

Contents

List of Contributors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . XIX

Chapter 1 - Membrane Transport in Biology (H. H. Ussing) ............... .

Chapter 2 - Passive Transport Processes (0. Sten-Knudsen)

A. Introduction . . . . . .

B. Fundamental Definitions I. Flux ....... . II. Types of Passive Transport

1. Diffusion 2. Migration . 3. Convection

III. Flux Equations 1. Migration Flux 2. Convection Flux 3. Diffusion Flux

a) Fick's Law . b) The Driving Force behind the Diffusion Process

4. Diffusion and Migration Proceeding Concurrently 5. Convection with Superimposed Diffusion

1

5

5

5 5 6 6 7 7 7 7 9 9

10 11 12 13

C. Diffusion Processes: Macroscopic Treatment 13 I. The Diffusion Equation ........ 14

1. Classification of Diffusion Processes 15 II. Stationary Processes in One Dimension 15

1. Steady-State Diffusion in a Plate 16 2. The Permeability Coefficient 17 3. Stationary Diffusion through Two Different Media 19

III. Time-Dependent Processes ............. 20 1. Kinetics of Exchange between Two Phases Separated by a Membrane. 21

a) One of the Phases is Infinitely Large .............. 21 (i) Outer Concentration Zero ................. 21 (ii) Outer Concentration Finite, Inner Concentration Initially

Zero ............................ 22

VIII Contents

b) Both Phases Comparable in Size ..... 23 c) Unidirectional Fluxes ..... ..... 26

2. Instantaneous Point Source (Green's Function) 27 a) Solutions of Diffusion Problems by Means of Green's Function 29

(i) Initial Uniform Distribution in the One Half-Space 30 (ii) The One Half-Space is Separated by an Impermeable Wall 30 (iii) The Presence of an Absorbing Barrier 31 (iv) Variable Flux into the Half-Space 31

3. Diffusion Out of a Plate. . . . . . . . . . . . 32 a) Concentration Profiles .......... 32 b) The Time Constant for the Exchange of the Mean Concentration 35

4. Establishing the Stationary Concentration Profile 37

D. Diffusion Processes: Microscopic Aspects 40 I. Brownian Movements . . . . . . . 40 II. Smoluchowski's Treatment 41

1. Statistical Interpretation of the Diffusion Equation 41 2. Random Walk in One Dimension . . . 42 3. The Einstein-Smoluchowski Equation 45

III. Random Walk and Fick's Law (Einstein) 46 IV. The Smoluchowski Equation 48 V. Kramers' Equation ....... 51 VI. Diffusion Coefficient and Mobility 51

1. Einstein's Relation ... 52 2. Einstein-Stokes'Relation . . . 53

E. Diffusion and Superimposed Convection 53 I. The Equation of Motion . , . . . . 54 II. Steady-State Concentration Profile 55

1. Stationary Transport through a Membrane 55 a) Determination of the Flux . . . . . . 55 b) Unidirectional Fluxes and Flux Ratio 56 c) The Concentration Profile 57

F. Electrodiffusion . . . . . . . . . . 58 I. Conductance......... 59 II. The Nemst-Planck Equations 61

1. Various Equivalent Forms 61 2. The Poisson Equation . 63

a) Electroneutrality 64 b) The Constant Field 65

III. Membrane Equilibrium . 65 1. Nonosmotic Equilibrium 65

a) The Nemst Equation 65 b) Equivalent Electrical Circuit for the Ion-Selective Membrane 67

2. Donnan Equilibrium ..................... 68 a) Thermodynamic Treatment . . . . . . . . . . . . . . . . 68 b) Concentration and Potential Profiles between the Phases

(The Poisson-Boltzmann Equation) . 73 IV. Diffusion Potentials ............ 80

1. Charging Time and Redistribution Time 82 2. The Henderson Regime . . . . . . . . . 84

Contents IX

3. The Planck Regime . . . . . . . . . . . . . . . . . . . . . 85 a) Planck's General Relations . . . . . . . . . . . . 86 b) The Electrical Equivalent Circuit for the Planck Regime 87 c) Planck's Expression for the Diffusional Potential 90

V. Electrodiffusion through Membranes 91 1. Single Salt . . . . . . . . 91

a) Diffusion Potential 91 b) Membrane Resistance . . . 92 c) Equivalent Electrical Circuit 94 d) Electroneutrality ..... 95

2. Ion-Selective Membrane ... 96 3. Membrane Separating Electrolytes Having a Common Ion 98

a) Aux Ratio . . . . . . . . . . . . . . . . . . . . . . . 98 b) The Goldman Regime . . . . . . . . . . . . . . . . . 99

(i) The Separate Ionic Currents and the Diffusion Potential 100 (ii) Total Membrane Current and Membrane Potential 102 (iii) Concentration Profiles and Membrane Potential 104 (iv) Ionic Conductances and Membrane Potential 106

c) Equivalent Electrical Circuits 109

Acknowledgements List of Symbols

References

Chapter 3 - Interpretation of Tracer Fluxes (H. H. Ussing) ..... .

A. Introduction . . . . . . . . . .

B. Fundamental Concepts

C. Tracer Permeability Coefficients I. Measurement of Tracer Permeability Coefficients II. Multicompartment Systems .......... .

D. The Concept of Unidirectional Aux I. Unidirectional Auxes II. Isotope Effects ....... . III. Associated Unidirectional Auxes IV. The Relation of Tracer Auxes to Active and Passive Transport V. Effects of Membrane Potentials on Ionic Auxes VI. Exchange Diffusion ............ . VII. Limitations for Integration of Aux Equations

E. Aux Ratio Analysis ........... . I. The Aux Ratio Equation ..... . II. Derivation of the Aux Ratio Equation III. Estimation of Electrochemical Potential Differences IV. The Short-Circuiting Method ....... . V. Aux Ratio with Solvent Drag ....... . VI. Solvent Drag on Non-Electrolytes and Water VII. Solute-Solute Interactions . . . . . . . . . . VIII. Meaning of the Term "Interaction" ..... IX. Interpretation of Deviations from the Aux Ratio Equation

110

110

112

115

115

115

116 116 117

118 118 118 119 119 120 120 121

122 122 123 126 127 127 130 131 131 132

X Contents

F. Examples ............ 133 I. The Short-Circuited Frog Skin 133 II. Single-File Diffusion 137 III. Solvent Drag Effects 138

G. Concluding Remarks 139

References

Chapter 4 - Nonequilibrium Thermodynamics of Isotope Flow through Membranes (F. A. Sauer) ..

A. Introduction . . . . . . .

B. The System: Definitions and Mathematical Techniques

C. Nonthermodynamic Considerations of Isotope Flow

D. The Nonequilibrium Thermodynamic Approach

E. Applications to Model Systems

F. Summary

Acknowledgements

List of Symbols

References

Chapter 5 - Use of Microelectrodes for Measurement of Membrane Potentials (D. V. Lassen and B. E. Rasmussen)

A. Introduction . . . . . . . . . . .

B. Principles of Bioelectric Recording I. Electrode Chains and Junction Potentials II. Comments on Electronic Equipment

C. The Glass Capillary Microelecirode . . . . . . I. The Suspension Effect . . . . . . . . . . II. Diffusion Regime of the Microelectrode Tip

D. Potential Recording with Microelectrodes . . . . I. Penetration of the Cell Membrane . . . . . II. Microelectrodes and Leaks in the Membrane

E. Epilogue . . . .

Acknowledgements

References

Chapter 6 - Chemical Composition of Membranes (D. J. Hanahan)

A. Introduction . . . . . . . . . . . . . . . . . . . . . .

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148

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167

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169

169

170 171 175

178 181 183

190 190 194

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202

202

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Contents

B. Some General Observations on Erythrocyte Composition I. General Composition II. Ion Composition . . . . . . . . . III. Age-Related Patterns ..... .

1. Intact Mixed-Age Erythrocytes 2. Age (density)-Separated Erythrocytes

C. Comments on Major Components of the Erythrocyte Membrane I. Lipid: an Appraisal of Composition and Orientation or Localization

1. Some Structural Features of the Erythrocyte Lipids a) Neutral Lipid b) Phospholipids .. . . . . . . . . . . . . . . . c) Sphingoglycolipids ............. .

XI

.206 206 206 208 208 210

214 215 216 216 216 217

2. Observations on Types of Phospholipids Present in Human, Cow, and Pig Erythrocyte . . . . . . . . 217 a) Fatty Acid Composition . . . . . . . . 218 b) Positioning of Fatty Acids . . . . . . . 219 c) Importance of Fatty Acid Composition 220

3. Localization of Lipids in Membranes: A Compositional Study of a Different Type ...................... . a) Some General Observations ................ . b) Use of Enzymes as Probes for Location of Lipids in Membranes

4. Summary of Observations on Phospholipase Action on Erythrocytes a) Phospholipase Az b) Phospholipase C . . . . . . . . . . . . . . . . . . . . . . . . c) Sphingomyelinase . . . . . . . . . . . . . . . . . . . . . . . d) Combined Activity of Phospholipase C and Sphingomyelinase

.221 222 223 224 224 225 225 225

5. Development of the Concept of Asymmetric Location of Phospholipids in Membranes . . . . . . . . . . . . . . . . . . . 225

6. On the Validity of the Lipid Asymmetry Proposal . 226 7. Summary Statement . 231

II. Protein Composition . . . . . . . . . . . . . . . . 231 1. General Comments . . . . . . . . . . . . . . 232 2. Nature of Polypeptide Patterns on SDS-PAGE 233

a) Specific Protein Components Revealed by SDS Gel Electrophoresis ................... 233

b) Studies on "Spectrin" of the Human Erythrocyte Membrane 234 c) Observations on the Ox (Bovine) Erythrocyte Polypeptide

Heterogeneity . . 235 3. Summary Statement 236

References

Chapter 7 - Genetic Detennination of Membrane Transport (c. W. Slayman)

A. Introduction . . . . . . . . . . . .

B. Microorganisms .. . . . . . . . . I. Isolation of Transport Mutants II. Kinds of Genetic Analysis . . . III. Examples of the Use of Genetic Analysis

. 236

. 239

.239

.240 240 241 243

XII Contents

C. Higher Organisms . . . . . I. Cystinuria...... II. HK/LK Erythrocytes

D. Cultured Somatic Cells I. Methods for Selecting Transport Mutants II. The Kinds of Genetic Information that can be Obtained III. Ouabain-Resistant Mutants

E. Conclusions

References

Chapter 8 - Mechanisms of Ion Transport and ATP Fonnation (E. Racker) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

A. Translocation of Protons by the Oxidation Chain of Mitochondria and Chloroplasts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . I. The Chemiosmotic Mechanism of Mitchell ........... . II. Asymmetry of Oxidation Chain of Mitochondria and Chloroplasts III. The Coenzyme Q Cycle .......... .

B. The Translocation of Protons by Bacteriorhodopsin I. Proton Movements and A TP Formation . . . II. Mechanism of Proton Translacotion . . . . .

C. The Translocation of Protons by the Oligomycin- or Dicyclohexylcarbodiimide­Sensitive ATPase of Mitochondria, Chloroplasts and Bacteria ..... I. Proton Movements and A TP Formation . . . . . . . . . . . . . . II. Properties of the Isolated Oligomycin-Sensitive ATPase Complex

1. The Water-Soluble ATPase ............. . 2. The Oligomycin-Sensitivity Conferral Protein (OSCP) 3. The Heat-Stable Coupling Factor F6 (Fe2) 4. Coupling Factor 2 (F2) •••••••••••..

III. Model of the Proton Pump and its Mode of Action 1. The Mitchell Hypothesis .......... . 2. The Phosphoenzyme Intermediate Hypothesis . 3. The Boyer-Slater Hypothesis ......... .

D. Translocation of Calcium by the ATPase Complex of Sarcoplasmic Reticulum I. Properties of the Pump ......... . II. Properties of the Ca + + -ATPase Complex. . . . . . . .

1. Latency of the Ca++-ATPase ............ . 2. Structural Properties of the Ca++-ATPase Complex. 3. Catalytic Properties of the Ca++ -ATPase Complex .

II. The Reconstituted Pump and its Mechanism of Action

E. Translocation of Sodium and Potassium Ions by the ATPase Complex of the Plasma Membrane . . . . . . . . . . . . . . . . . I. Properties of the Pump .......... . II. Properties of the Na+-K+-ATPase Complex

1. Latency of the ATPase ......... . 2. Structural Properties of the Enzyme Complex 3. Catalytic Properties of the Enzyme Complex

III. The Reconstituted Pump

F. Concluding Remarks

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· 254

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· 261 · 261 .263

263 263

265 265 266 267 269 270 270 271 272 273 273

.274 274 276 276 276 277 277

281 281 282 282 282 283 284

.286

Acknowledgement

Abbreviations

Addendum

References

Contents

Chapter 9 - Membrane Immunological Reactions and Transport (P. K. Lauf) ........ .

A. Introduction: The Concept

XIII

287

287

287

287

291

291

B. Immunological Reactions and Membrane Transport Proteins 292 I. Introduction............... 292 II. Antibodies Against the Na+-K+-ATPase . . . . . . . 293

1. Properties of Antigens and Antibodies . . . . . . . 293 2. Sidedness of Binding and Immunological Effects on the

Na+ -K+ -ATPase and its Partial Reactions . . . . . . . 295 a) Sidedness of Binding ................ 295 b) Immunological Effects on the Na+-K+-ATPase Activity. 295 c) Effects of Partial Reactions of the Na+-K+-ATPase . . . 296

3. Immunological Alteration of Cation F1uxes in Resealed Ghosts . 297 4. Species and Organ Specificities of Immunological Reactions Involving

the Na+-K+-ATPase . . . . . . . . . . . . . . . . . . . . . 297 III. Antibodies Against Ca++-ATPase of Sarcoplasmic Reticulum ...... 298 IV. Conclusion ................................. 299

C. Immunological Reactions at the Outer Membrane Surface and Cation Transport in Erythrocytes 299 I. Introduction.......................... 299 II. Sheep Red Cells . . . . . . . . . . . . . . . . . . . . . . . . 301

1. Cation Transport, Genetics and Immunological Parameters 301 a) Cellular Cations and Genetics . . . . 301 b) Membrane Antigens and Genetics 302 c) Active and Passive Cation Transport 303 d) Ouabain Binding .......... 304 e) Na+ -K+ -ATPase. . . . . . . . . . . 304

2. The Effect of Antibodies on Cation Transport 305 a) Modification of Cation Pump and Leak F1uxes 305 b) Activation of the Na+-K+-ATPase . . . . . . 307 c) Correlation between Antigenic Sites and Na+ -K+ Pumps 308

3. Properties of the ML Surface Antigens and Antibodies 310 a) Antigens . . . . . . . . . . . . . . . . . . . . 310 b) Antibodies .................. 311

4. Developmental Aspects of Transport and Antigens 312 a) Red Cells of Newborn Sheep ......... 312 b) Stress-Induced Erythrocyte Regeneration . . . 313

III. Cation Transport Polymorphism and Antigenic Parameters in Red Cells of Ruminants Other than Sheep 315 1. Goat Red Cells ....... . . . . . . . . . . . . . 315

a) Cations and Antigens ............... 315 b) Cation Transport and its Modification by Antibody 315

2. Cattle Red Cells ................... 317

XIV Contents

IV. Human Red Cells .................. . 1. The Rhesus Antigen Complex and Cation Transport 2. The En(a)-Negative Red Cell as Physiological Model

V. Conclusion ...................... .

· 318 318

· 319 · 320

D. Membrane Immunological Reactions and Cation Transport in Lymphocytes and Other Cells ..... 321 I. Lymphocytes ......................... 321

1. Introduction ........................ 321 2. Cellular Differentiation and Membrane Surface Receptors

of Lymphocytes . . . . . . . . . . . . . . . . . . . . . . . 3. Cellular and Membrane Morphological and Biochemical Changes

Induced by Immunological Reactions in Lymphocytes .. . 4. Modification of Monovalent Cation Transport ......... .

a) General Aspect of the Effect of Immunological Reactions b) Cation Transport in the Absence of Immunological Reactions c) Cation Transport Changes Induced by Immunological Reactions

5. Requirement of Bivalent Cations for Lymphocyte Stimulation by Immunological Reactions

II. Tumor Cells ................... . 1. Introduction .................. . 2. Passive Permeability Changes Induced by Lectins

III. Conclusion.....

E. Summary and Prospectus

Acknowledgement

References

Chapter 10 - Membrane Receptors, Cyclic Nucleotides, and Transport (S. A. Rudolph and R. J. Lefkowitz)

A. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

B. Beta-Adrenergic-Receptor Binding in Avian and Amphibian Erythrocytes

C. Beta-Adrenergic-Mediated Transport Processes in Avian and Amphibian Erythrocytes

D. The Amphibian Bladder . . . .

E. Cholera Enterotoxin . . . . . .

F. The Superior Cervical Ganglion

G. Nicotinic Cholinergic Receptors

H. The Heart .......... .

J. General Comments and Conclusions

References

Chapter 11 - Permeability Properties of Unmodified Lipid Bilayer Membranes (0. S. Andersen)

A. Introduction . . . . . . .

321

.324 326 326 327 330

338 338 338 339 340

341

342

342

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· 366

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Contents

B. Lipid Bilayer Membranes I. Capacitance ... . II. Composition ... .

C. Transport Model and the Potential Energy Barrier I. The Transport Model II. Unstirred Layers . .

1. Stationary Fluxes . 2. Transient Fluxes 3. The Membrane-Solution Interface 4. Chemical Reaction in Unstirred Layers

III. Potential Energy of Ions Within Lipid Bilayers 1. The Born Energy . . . . 2. The "Image" Force . . . . . . . . . . . . . 3. Diffusion or Distortion? ......... .

IV. Potential Energy of Dipolar Molecules Within Lipid Bilayers V. Interfacial Potentials . . . . . . . .

1. Diffuse Double-Layer Potentials 2. Dipole Potentials . . . . .

VI. Hydrophobic Interactions . . VII. The Potential Energy Barrier

D. Permeability to Neutral Solutes I. Partition Coefficients

1. Nonpolar Solutes 2. Polar Solutes

II. Mobility . . . . . . . 1. Indirect Measurements

a) Microviscosity b) Walden's Rule . . .

2. Direct Estimates 3. Variation through the Membrane

III. Permeability . . . 1. H 20 .............. . 2. Organic Solutes . . . . . . . . .

IV. The Rate-Limiting Barrier for Solute Movement E. Ion Permeability . . . . . . . . .

I. Tracer Flux Measurements. . . . . II. Anion Permeability ....... .

1. Stationary Conductance Changes 2. Translocation through the Membrane Interior

a) The Transport Model ..... . b) Kinetics of Charge Translocation .... . c) Temperature-Dependence .... . d) Ion Translocation as a Function of Membrane Composition

III. Positive Ions . . . . . . . . . . . . . . . . . . . . . . . . IV. Interactions Among Ions Absorbed into Lipid Membranes

1. Space Charge-Limited Conductance 2. Blocking Phenomena . . . . 3. The Three-Capacitor Model

a) Charge Adsorption b) Charge Translocation

4. Discrete Charge Effects?

xv

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.373 373 375 376 376 378 378 379 379 380 384 385 385 387 388 389 390 392 393 393 394 395 395 395 395 396 396 397 397 399

.400

.402 403 404 404 406

.408 · 415 · 419 .419

425 426 427 428

.430 · 431 .432 .435

XVI

Acknowledgements

References

Contents

Chapter 12 - Carrier-Mediated Ion Transport Across Thin Lipid Membranes (G. Stark) ........ .

A. Introduction . . . . . . I. Carriers and Pores II. A Survey of Suggested Ion Carriers

B. Carriers of Hydrogen Ions

C. Macrocyclic Carriers . I. Neutral Carriers . . II. Charged Carriers

D. The Iodide-Iodine System

E. The Carrier-Transport Model I. Kinetic Analysis of the Carrier Model II. Valinomycin and Trinactin

F. Biological Implications

References

Chapter 13 - Channels in Black Lipid Films (J. E. Hall)

A. Introduction . . . . . . . . . . . . . . . . .

B. Basic Experiments . . . . . . . . . . . . . . I. Demonstration of Conductance by Pore II. Basic Conductance Characteristics

1. Steady-State Current-Voltage Curves 2. Conductance and Antibiotic Concentration 3. Kinetics of Conductance Development: Response to a Voltage Pulse

C. Advanced Experiments I. Introduction.......... II. Single-Step Experiments

1. The Probability Distribution 2. ElM and Hemocyanin: The Unit Event Explains High-Level

Conductance. . . . . . . . . . . . . . . . . . . 3. Noise Measurements and the Unit Conductance 4. Noise Measurements on Alamethicin . . . . . . 5. Noise Measurements on Monazomycin . . . . . 6. Compounds with Unknown Unit Events: Summary

III. Conductance and Ion Selectivity of Unit Channels 1. Introduction .................. . 2. Conductance and Selectivity of the Gramicidin Unit Event 3. Conductance and Selectivity of the Unit Events of ElM

and Hemocyanin ..................... . 4. Conductance and Selectivity of the Alamethicin Unit Event Levels

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Contents

IV. Time Course of the Unit Event V. Alteration of the Molecule and the Membrane

1. Effects of Membrane Composition .... 2. Alteration of the Pore Forming Molecule

D. Possible Molecular Mechanisms of Pore Formation

Acknowledgements

References

XVII

517 519 520

. 523

.525

529

529

Subject Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 533

List of Contributors

Olaf S. Andersen Cornell University, Medical College, Department of Physiology, 1300 York Avenue, New York, N.Y. 10021 / USA

James E. Hall Dept. of Physiology, California College of Medicine, University of California, Irvine, California 92717 / USA

Donald J. Hanahan The University of Texas, Health Science Center at San Antonio, Department of Biochemistry, 7703 Floyd Curl Drive, San Antonio, Texas 78284 / USA

Ulrik V. Lassen University of Copenhagen, August Krogh Institute, Zoophysiological Laboratory B, 13 Universitetsparken, DK-2100 Copenhagen

Peter K. Lauf Department of Physiology, Duke University Medical Center, Durham, North Carolina 27710 / USA

Robert J. Lefkowitz Duke University Medical Center, M 3325, Durham, North Carolina 27710 / USA

Efraim Racker Cornell University, Section of Biochemistry, Molecular and Cell Biology, Wing Hall, Ithaca, New York 14853 / USA

B. E. Rasmussen University of Copenhagen, August Krogh Institute, Zoophysiological Laboratory B, 13 Universitetsparken, DK-2100 Copenhagen

Stephen A. Rudolph Case Western Reserve University, School of Medicine, Dept. of Pharmacology, Cleveland, Ohio 44106 / USA

xx List of Contributors

Friedrich A. Sauer Max-Planck -Institut fUr Biophysik, Kennedyallee 70, 0-6000 Frankfurt/Main 70

c. W. Slayman Yale University, School of Medicine, Department of Human Genetics, 333 Cedar Street, New Haven, Conn. 06510 / USA

Gunther Stark Universitat Konstanz, Fachbereich Biologie, D-7750 Konstanz

Ove Sten-Knudsen University of Copenhagen, Panuminstituttet, Department of Biophysics, Blegdamsvej 3 C, DK-2200 Copenhagen N

Hans H. Ussing University of Copenhagen, Institute of Biological Chemistry A, 13 Universitetsparken, DK-2100 Copenhagen