Table Of ContentCELL MEMBRANE
TRANSPORT
Principles and Techniques
CELL MEMBRANE
TRANSPORT
Principles and Techniques
by Arnost Kotyk and Karel Janacek
Laboratory for Cell Membrane Transport
Institute of Microbiology
Czechoslovak Academy of Sciences
Prague, Czechoslovakia
In collaboration with the Staff of the Laboratory
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PLENUM PRESS . NEW YORK - LONDON . 1970
Library of Congress Catalog Card Number 71-107538
ISBN-13: 978-1-4684-0720-4 e-ISBN-13: 978-1-4684-0718-1
001: 10.1007/978-1-4684-0718-1
© 1970 Plenum Press, New York
Softcover reprint of the hardcover 1s t edition 1970
A Division of Plenum Publishing Corporation
227 West 17th Street, New York, N.Y. 10011
United Kingdom edition published by Plenum Press, London
A Division of Plenum Publishing Corporation, Ltd.
Donington House, 30 Norfolk Street, London W.C. 2, England
All rights reserved
No part of this publication may be reproduced in any form
without written permission from the publisher
To students of
flow across barriers - not only biological
PREFACE
It is not a particularly rewarding task to engage in writing a book on a
subject which is undergoing a rapid and potentially revolutionary develop
ment, but, on the other hand, the investigation of transport of substances
into and out of cells has reached a stage of maturity or at least of self
realization and this fact alone warrants a closer examination of the subject.
No one will doubt at present that the movement-mostly by selective
translocation-of substances, ranging from hydrogen ions to deoxyribo
nucleic acids, across the cell-surrounding barriers represents one of the
salient features of a living cell and that, if we are permitted to go so far,
the cessation of the selective transport processes might be considered as
the equivalent of cell death. Hardly anybody will question the premise
that cell and tissue differentiation within the ontogenetic development of
an organism is closely associated with properties of the outer cell face.
Perhaps no serious scholar will attempt to refute the concept that mem
branes with characteristic morphology and composition represent the ar
chitectural framework for the whole cell. And probably no experienced
biologist will raise objections to the belief that many physiological processes,
like nervous impulse conduction and other electrical phenomena of cells
and tissues or their volume changes, are associated with membrane-regulated
shifts of ions and molecules.
Still, perhaps because all these observations comprise a border discipline
par excellence, textbooks of biochemistry hardly ever mention the problem
of biological transport, textbooks of physiology deal at most with the ion
and water movement in the kidney, and textbooks of biophysics (the few
there are) treat membrane phenomena and ion distribution in cells rather
one-sidedly.
Bringing together all the pertinent information-an awesome task-is
thus imperative at this stage of experimental research. However, collating
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viii Preface
the diversified pieces of evidence within a system has proved to be extremely
difficult, apparently owing to our notions of systems acquired either in
physics or in chemistry or in biology. It is not intended to be an over
estimation of the magnitude of the work facing the membrane biologist
(or chemist or physicist?) if we state that none of the classical frameworks
will fit here flawlessly. Therefore, we were compelled to resort to a certain
compromise when designing the layout of this book. It includes a physico
chemical treatment arranged according to the degree of complexity of
transport models, as well as one based on the type of biological objects
examined. We believe that this dualism is not detrimental to the usefulness
of the book as each of the sections contains a different type of information
and, from the viewpoint of supplying data to the reader, we feel that it is
actually of greater practical value.
While we realize that this lack of a unitary system is a handicap for
our present effort it will only add to the merit of those who, perhaps in
the near future, will be able to propose a neat and orderly classification of
transport processes based on their molecular parameters, a challenge to the
enzymologist and to the protein chemist.
This is not the first book on biological transport ever written, a mono
graphic treatment of one or more aspects of transport or cell membranes
having appeared several times before, the more important and recent ones
of these being included in the bibliography at the end of this book. How
ever, this book has been written not only to summarize what we feel to be
important experimental evidence and theories attempting to account for it
but also to provide the reader with an insight into the technical means that
are available for transport studies. And it is this section of the book which
we think might be especially useful to those aspiring to study transport
phenomena in various types of cells.
It was our intention to include in the text all the physical concepts
underlying the more sophisticated phenomena and we did our best to use
uniform symbols throughout although certain inconsistencies were unavoid
able because of traditionally used symbols in different iteatments of the
subject (such as the symbol c as well as s used for solute concentration).
We attempted to include, wherever appropriate, practical examples to
illustrate either the theories or the techniques.
The 26 chapters of this book are divided into 5 sections so as to em
phasize the different approaches employed. The first section deals with the
morphological and chemical aspects of membranes and it was included
to present the student with a physical picture of the object of the subsequent,
largely abstract, considerations.
Preface ix
The second section covers most, if not all, of the kinetic concepts and
theories involved in membrane transport. It starts necessarily with a chapter
on diffusion of substances and migration of ions in a homogeneous medium,
to proceed to the transport of substances in anisotropic systems and to the
metabolically linked types of transport. The section also includes a brief
treatment of special types of transport for which the detailed kinetics are
not known, viz., pino- and phagocytosis.
The third section is one which will probably become obsolete most
rapidly as it presents the rather meager but fast accumulating evidence on
the molecular properties and plausible models of the membrane transport
systems.
The fourth section contains technical descriptions of methods and
basic equipment used in transport studies, together with practical examples
illustrating their application.
The fifth section is a more or less comparative treatment of the transport
information as has accrued from studies of different cell types.
The writing of the various chapters of the book was accompanied by
frequent mutual consultations and the whole text was revised several times
to avoid major inconsistencies and duplicity. If errors and shortcomings
are found by readers of the book their criticism should be addressed to
A.K. for chapters 1,3.2.,5.1.2.,5.2., 7, 8, 9, 10, 11, 15, 16, 17, 19, and 26,
and to K.J. for chapters 2, 3.1., 4, 5.1.1., 6, 12, 13, 22, 24.1, 24.2., 24.3.,
and 24.4.
It is a pleasure to acknowledge the factual and moral support for
writing this book shown by our colleagues and friends. Among those who
actively contributed, greatest credit goes to Dr. Renata Rybova who wrote
chapters 18, 20, and 21; to Dr. Jifina Kolinska who contributed greatly
by writing chapters 23, 24.6., 24.7., and 24.8.; to Dr. Milan HOfer who
wrote the chapter on mitochondria (25); to Dr. Rudolf Metlicka and Mr.
Ivan Benes who wrote chapter 14 on the use of artificial membranes; and
to Dr. Stanislav Janda who contributed by sections 23.4.2 and 24.5.
Other collaborators helped us greatly by calling our attention to pub
lications that had escaped our attention (special credit to Dr. Jaroslav
Horak) and by discussing the text with us.
We are greatly indebted to Dr. Jifi Ludvik who heads the Laboratory
of Electron Microscopy of this institute who supplied a number of original
electron microphotographs. Our thanks are due to a number of assistants
for typing parts of the manuscript, drawing some of the figures, and photo
graphing others.
It would be most ungrateful not to mention here the debt we owe to
x Preface
Prof. Arnost Kleinzeller, now at the University of Pennsylvania, who in
troduced us into the realm of transport studies and, last but not least, to
Prof. Ivan Malek who, as head of the Institute of Microbiology of the
Czechoslovak Academy of Sciences, showed profound understanding for
the problems of membrane transport when he permitted a whole depart
ment of his institute to concentrate on biological transport phenomena.
Prague, July 1969 Arnost Kotyk
Karel Janacek
Laboratory for Cell Membrane Transport
Institute of Microbiology
Czechoslovak Academy of Sciences
CONTENTS
Structural Aspects
1. Composition and Structure of Cell Membranes .............. 3
1.1. Cell Envelopes and Membranes ....................... 3
1.2. Chemical Composition of the Plasma Membrane . . . . . . . . 12
1.3. Fine Structure of the Plasma Membrane . . . . . . . . . . . . . . . . 15
Kinetic Aspects
2. Transport in Homogeneous Liquid Phase ................... 27
2.1. Mass Flow and Diffusion ............................ 27
2.1.1. Introductory Definitions .. . . . . . . . . . . . . . . . . . . . . . 27
2.1.2. The Laws of Diffusion ........................ 28
2.2. Migration and Electrodiffusion of Ions . . . . . . . . . . . . . . . . . 39
3. Passive Membrane Transport of Nonelectrolytes ............. 55
3.1. Permeation by Simple Diffusion ...................... 55
3.2. Mediated Diffusion and Related Phenomena . . . . . . . . . . . . 61
3.2.1. General considerations ........................ 61
3.2.2. Model I ..................................... 63
3.2.3. Model II .................................... 75
3.2.4. Model III ................................... 79
3.2.5. Model IV . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 82
3.2.6. Inhibition of Mediated Diffusion ............... 86
4. Permeation of Ions and Water. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 91
4.1. Permeation of Ions and Membrane Potentials. . . . . . . . . . . 91
4.1.1. Introduction ................................. 91
4.1.2. Equilibrium of Ions across Membranes 92
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