Table Of ContentIn memory
of
Dr. Han de Man
(1924-1976)
Manroalaöi Cell
(^[®ΙΠΠ3^[Γ^Ο®3
V O L U ME T WO
The Diversity of Membranes
Edited by
G. A. Jamieson Ph.D., D.SC.
Research Director
American Red Cross Blood Research Laboratory
Bethesda, Maryland, USA
and
Adjunct Professor of Biochemistry
Georgetown University Schools of Medicine and Dentistry
Washington, DC, USA
and
D. M. Robinson Ph.D.
Professor of Biology, Georgetown University
and
Member, Vincent T. Lombardi Cancer Research Center
Georgetown University Schools of Medicine and Dentistry
Washington, DC, USA
B U T T E R W O R T HS
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First published 1977
© Butterworth & Co (Publishers) Ltd 1977
ISBN 0 408 70723 2
Library of Congress Cataloging in Publication Data
Main entry under title:
Mammalian cell membranes.
Includes bibliographical references and index.
CONTENTS: v. 1. General concepts, v. 2. The
diversity of membranes, v. 3. Surface membranes of
specific cell types, v. 4. Membranes and cellular functions.
1. Mammals—Cytology. 2. Cell membranes.
1. Jamieson, Graham Α., 1929- II. Robinson,
David Mason, 1932- [DNLM: 1. Cell membrane.
2. Mammals. QH601 M265]
QL739.15.M35 599'.08'75 75-33317
ISBN 0-408-70723-2 (v. 2)
Filmset and printed Offset Litho in Great Britain by
Cox & Wyman Ltd, London, Fakenham and Reading
Contributors
RODERICK A. CAPALDI
Department of Biology and Institute of Molecular Biology, University of
Oregon, Eugene, Oregon 93703, USA
ROBERT P. DONALDSON
Department of Biochemistry, Michigan State University, East Lansing,
Michigan 48824, USA
P. EMMELOT
Department of Biochemistry, Antoni van Leeuwenhoek Laboratory, The
Netherlands Cancer Institute, Amsterdam, The Netherlands
p. FAVARD
Centre National de la Recherche Scientifique, Centre de Cytologie Expéri-
mentale, 94200 Ivry-sur-Seine, France
D. J. FRY
Department of Anatomy, Medical Sciences Institute, University of Dundee,
Dundee DDI 4HN, Scotland
J. J. GEUZE
Center for Electron Microscopy, Medical Faculty, University of Utrecht,
Nicolas Beetsstraat 22, Utrecht, The Netherlands
NICHOLAS A. KEFALIDES
Departments of Medicine and Biochemistry, University of Pennsylvania
and Philadelphia General Hospital, Philadelphia, Pennsylvania 19104, USA
M. F. KRAMER
Laboratory for Histology and Cell Biology, Medical Faculty, University of
Utrecht, Nicolas Beetsstraat 22, Utrecht, The Netherlands
c.
j. H. DE MAN
Laboratory for Pathology, University of Leyden, Wassenaarseweg 62,
Leyden, The Netherlands
F. A. RAWLINS
Centro de Biofisica y Bioquimica, Instituto Venezolano de Investigaciones
Cientificas (IVIC), Apartado 1827, Caracas, Venezuela
CONTRIBUTORS
PETER SATIR
Department of Physiology-Anatomy, University of California, Berkeley,
California 94720, USA
N. E. TOLBERT
Department of Biochemistry, Michigan State University, East Lansing,
Michigan 48824, USA
B. G. UZMAN
Centro de Biofisica y Bioquimica, Instituto Venezolano de Investigaciones
Cientificas (IVIC), Apartado 1827, Caracas, Venezuela
G. M. VILLEGAS
Centro de Biofisica y Bioquimica, Instituto Venezolano de Investigaciones
Cientificas (IVIC), Apartado 1827, Caracas, Venezuela
ROBERT WATTIAUX
Facultés Universitaires Notre-Dame de la Paix, Laboratoire de Chimie
Physiologique, 66 rue de Bruxelles, 5000 Namur, Belgium
Preface
This series on 'MAMMALIAN CELL MEMBRANES' represents an attempt to bring
together broadly based reviews of specific areas so as to provide as compre-
hensive a treatment of the subject as possible. We sought to avoid producing
another collection of raw experimental data on membranes, rather have we
encouraged authors to attempt interpretation, where possible, and to express
freely their views on controversial topics. Again, we have suggested that
authors should not pay too much attention to attempts to avoid all overlap
with fellow contributors in the hope that different points of view will provide
greater illumination of controversial topics. In these ways, we hope that
the series will prove readable for specialists and generalists alike.
The first volume, entitled General Concepts, served to introduce the subject
and covered the essential aspects of physical and chemical studies which
have contributed to our present knowledge of membrane structure and
function. This, the second volume, is called The Diversity of Membranes and
addresses itself to specific types of intra- and extracellular membranes, while
the third volume, Surface Membranes of Specific Cell Types, as its title
indicates, will review the knowledge that we have of the surface membranes
of the various cell types which have been studied in any detail to this time.
Membranes and Cellular Functions will be covered in Volume 4, which will
concern ultrastructural, biochemical and physiological aspects. Since the
cell surface represents the point of interaction with the cellular environment,
Volume 5, entitled Responses of Plasma Membranes, deals with the way in
which external influences are mediated by the plasma membrane.
As editors, our approach to our responsibilities has been rather permissive.
With regard to nomenclature and useful abbreviations, we have used 'cell
surfaces' and 'plasma membranes' where appropriate rather than 'cell
membranes' since this last is nonspecific. Both British and American usage
and spelling have been utilized depending upon personal preference of the
authors and editors with, again, no attempt at rigid adherence to a particular
style.
While the title of the series is 'MAMMALIAN CELL MEMBRANES', we have
encouraged authors to introduce concepts and techniques from non-
mammalian systems which may be useful in their application to eukaryotic
cells. The aim of this series is to provide a background of information and,
hopefully, a stimulation of interest to those investigators working in, or
about to enter, this burgeoning field.
Finally, the editors would like to acknowledge the dedication and resource-
fulness of their secretary and editorial assistant, Mrs Alice R. Scipio, in the
coordination and preparation of these volumes.
G. A. JAMIESON
D. M. ROBINSON
1
The organization of the plasma
membrane of mammalian cells :
structure in relation to function
P. Emmelot
Department of Biochemistry, Antoni van Leeuwenhoek Laboratory,
The Netherlands Cancer Institute, Amsterdam
1.1 INTRODUCTION
Today the study of membranes is flourishing and extending its scope to
many biological problems, physiological as well as pathological (Wallach,
1973). The present chapter is limited to some aspects of plasma membrane
structure and function, and tries to outline the possible significance for
membrane function of the organizational disposition of membrane con-
stituents. One is faced here with the intrinsic difficulty of demonstrating
whether a particular component or activity being selected for measurement
is the cause or the result of the biological process under consideration.
However, temperature-sensitive mutant cell lines (Willingham, Carchman
and Pastan, 1973) and transforming viruses (Otten et al, 1972) have been
of considerable help in this respect.
Mammalian cells are enclosed by the plasma membrane, sometimes called
the 'cell envelope', 'cell surface membrane', 'plasmalemma', or 'cell mem-
brane'. This membrane separates the cell interior from the exterior and, in
this manner, behaves in many respects as a dynamic organelle rather than
as a passive sieve or a static border.
The histogenetic integration of cells into tissue imposes 'polarity' which
is also expressed at the plasma membrane level. Regional specializations
occur for transport purposes, in separate areas according to whether uptake
or excretion occurs, for example in blood front and bile space lining mem-
branes in the liver. Intercellular contact is found at membrane junctions,
which have adhesive, communicative and barrier functions in, for example,
desmosomes, and gap and tight junctions (McNutt and Weinstein, 1973).
1
2 ORGANIZATION OF THE PLASMA MEMBRANE
It follows that the plasma membrane is not only subject to, but is also involved
in, the positional control of cells in recognition and intercellular contact.
Breakdown of control in this area generates free cells; this may be one of
the requirements for metastasis of cancer cells.
Cells derived from solid tissue which have been subsequently grown as
free cells, either in vitro or in vivo (for example, ascites tumor cells) lack the
principal phenotypic expressions of the histogenetic relations of their parent
cells in situ such as the junctional membrane complexes and other local
differentiations of the plasma membranes. However, the genetic potential
to form these specialized membrane structures may be variously retained,
as shown by the solid growth of ascites tumor cells after subcutaneous
transplantation, and by island formation of certain strains of these cells.
Other membrane expressions, such as the topographical contributions of
sialic acid, may also differ according to whether these cells are grown in
ascites or in solid form (Cook, Seaman and Weiss, 1963).
Dramatic changes to the trilamellar unit membrane result from regional
specializations such as tight and gap junctions (McNutt and Weinstein,
1973). A trilamellar structure featured in the earliest membrane models
(Gorter-Grendel, Davson-Danielli, Robertson), but models have been con-
stantly changing in the last 15 years to incorporate new structures and ideas
(Finean, 1972). The present favorite is the fluid-mosaic membrane model
(Singer and Nicolson, 1972). These models depict the arrangement of the
molecular species which compose the membrane element proper.
However, among the regional specializations of plasma membranes are
the globules of diameter 5-6 nm, which are present on differentiated areas
of plasma membranes of certain cells and appear to be related to transport
mechanisms in the membranes {see p. 38).
Glycoprotein may be integrated within the membrane (e.g. of the liver
cell surface) but may also extend quite far from the cell surface to form a
glycocalyx (Bennett, 1963), as is the case in intestinal cells (Ito, 1965), which
may form a filamentous network or 4fuzz' (Parsons and Subjeck, 1972) that
traps molecules and immobilizes water. Thus, there may be a long-range
ordering of water molecules around these parts of the cell surface, causing
local high viscosity (Schultz and Asunmaa, 1970; Drost-Hansen, 1971).
Bundles of 4-6 nm microfilaments are found in the cell cortex immedi-
ately below, if not in contact with, the cytoplasmic side of the plasma
membrane. There is now suggestive evidence that these ectoplasmic micro-
filaments actively participate in a number of membrane processes. Thus,
one arrives at what has been called the 'greater membrane' (Revel and Ito,
1967), which also contains the structures associated with the membrane
element.
As well as exerting positional control, the cell surface is also involved in
growth control. This conclusion has been drawn from the many experiments
in vitro on cell growth behavior, especially the 'contact inhibitions of move-
ment and growth' (Emmelot, 1973). It seems not unlikely that positional
and growth controls are related, some form of contact being required for
growth control.
Finally, the cell surface functions in immunological control or surveillance
and contains a group of special glycoproteins, the transplantation antigens,
which are instrumental in the rejection of a tissue graft in a noncompatible
ORGANIZATION OF THE PLASMA MEMBRANE 3
host. Similarly, chemical changes in determinants of the cell surface or
production of non-self components by mutation or viral infection (e.g. in
tumor cells) may cause the immunological apparatus to become active.
Immunologically competent cells contain cell surface expressions which
sense or recognize the non-self expression on their target cells. Recognition
of non-self may thus be related in mechanism to an aspect of positional
control, viz. the recognition of self between cells of a tissue. The difference
lies in the reaction to the recognition, leading to target-cell destruction in
the case of immunocompetent cells and to histogenetic integration or dis-
crimination of nonimmunocompetent cells.
Positional and immunological as well as growth controls serve to maintain
and protect the organism and its parts, and this establishment of barriers
seems—both literally and figuratively—to be a function of cell surface
interactions.
1.2 CHEMICAL COMPOSITION AND MEMBRANE
ARCHITECTURE
Plasma membranes consist of lipids, proteins and carbohydrates, the latter
contained in glycoproteins and glycolipids, varying in amount according to
cell type. Small amounts of both RNA and DNA have been detected in
various plasma membrane preparations but neither their function nor
whether they are genuine components of the plasma membrane has been
definitely established; most likely all DNA, and at least some of the RNA,
encountered in certain preparations represents contamination (Emmelot and
Bos, 1972).
The membrane constituents are combined into a three-dimensional,
supramolecular arrangement, held together mainly by noncovalent bonds,
of lateral continuity and exhibiting a certain geometrical width in which the
lipid bilayer and the attendant proteins are accommodated. X-ray diffraction
analyses have shown that the membrane width may be appreciably greater
than that observed by electron microscopy (Finean et al., 1968). Osmium
tetroxide has recently been shown to be a poor fixative for erythrocyte
membranes since it may remove membrane protein, whereas fixation with
5% glutaraldehyde preserves most of the membrane protein and yields a
membrane of width 16 nm (McMillan and Luftig, 1973).
The lipid bilayer, which forms a barrier to the free flow of solutes, is now
a reasonably well established feature of the plasma membrane (Wilkins,
Blaurock and Engelman, 1971; Coleman, 1973), though modified from the
classical model of Davson and Danielli. In that model a lipid bilayer was
coated by a sheet of protein on both sides. In the present fluid-mosaic mem-
brane model (Singer and Nicolson, 1972), proteins occur both within the
membrane and on its two faces. This concept of a lipid continuum locally
interrupted by protein intercalations reconciles the two earlier and distinct
concepts of membrane structure, namely the concepts of bilayer and globular
organization (Benedetti and Emmelot, 1968).
The general design of the plasma membrane is that of a sheet with two
hydrophilic sides, one facing the outside and the other the inside of the cell,
and a lipophilic, hydrophobic interior. The cell surface is generally negatively
Figure 1.1 isolated rat-liver plasma membranes stained with colloidal iron hydroxide. The
electron-dense granules are restricted to the outer leaflet of the membranes (inset). Junctional
complexes (brackets) are not stained. The bars, in both main figure and inset, represent 0.1 μχη.
(From Benedetti and Emmelot, 1967, courtesy of the Company of Biologists)
