The Fluid Mosaic Model of the Structure of Cell Membranes

Author(s): S. J. Singer and Garth L. Nicolson

Source: Science, New Series, Vol. 175, No. 4023 (Feb. 18, 1972), pp. 720-731
Published by: American Association for the Advancement of Science
Stable URL: http://www.jstor.org/stable/1733071
Accessed: 16-09-2016 23:50 UTC

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 experimental evidence in terms of the
model; and (v) to show that the fluid
mosaic model suggests new ways of
The Fluid Mosaic Model of the thinking about membrane functions and
membrane phenomena.

Structure of Cell Membranes

Thermodynamics and
Membrane Structure
Cell membranes are viewed as two-dimensional solutions
The fluid mosaic model has evolved
of oriented globular proteins and lipids.
by a series of stages from earlier ver-
sions (1-4). Thermodynamic considera-
tions about membranes and membrane
S. J. Singer and Garth L. Nicolson
components initiated, and are still cen-
tral to, these developments. These con-
siderations derived from two decades
of intensive studies of protein and nu-
Biological membranes play a crucial considerable detail several models of cleic acid structures; the thermodynamic
role in almost all cellular phenomena, the gross structural organization of principles involved, however, are per-
yet our understanding of the molecular membranes, in terms of the thermody- fectly general and apply to any macro-
organization of membranes is still rudi- namics of macromolecular systems andmolecular system in an aqueous en-
mentary. Experience has taught us, how- in the light of the then available ex- vironment. These principles and their
ever, that in order to achieve a satisfac- perimental evidence. From this analysis,application to membrane systems have
tory understanding of how any biologi- it was concluded that a mosaic struc- been examined in detail elsewhere (1)
cal system functions, the detailed ture of alternating globular proteins and are only summarized here. For our
molecular composition and structure of and phospholipid bilayer was the only present purposes, two kinds of non-
that system must be known. While we membrane model among those analyzed covalent interactions are most impor-
are still a long way from such knowl- that was simultaneously consistent with tant, hydrophobic (5) and hydrophilic
edge about membranes in general, prog-thermodynamic restrictions and with all (1). By hydrophobic interactions is
ress at both the theoretical and experi- the experimental data available. Since meant a set of thermodynamic factors
mental levels in recent years has brought that article was written, much new evi-that are responsible for the sequester-
us to a stage where at least the gross dence has been published that strongly ing of hydrophobic or nonpolar groups
aspects of the organization of the pro- supports and extends this mosaic model. away from water, as, for example, the
teins and lipids of membranes can be In particular, the mosaic appears to be immiscibility of hydrocarbons and
discerned. There are some investigators, a fluid or dynamic one and, for many water. To be specific, it requires the
however, who, impressed with the great purposes, is best thought of as a two- expenditure of 2.6 kilocalories of free
diversity of membrane compositions and dimensional oriented viscous solution. energy to transfer a mole of methane
functions, do not think there are any In this article, we therefore present and from a nonpolar medium to water at
useful generalizations to be made even discuss a fluid mosaic model of mem- 25?C (5). Free energy contributions of
about the gross structure of cell mem- brane structure, and propose that it this is magnitude, summed over the many
branes. We do not share that view. We applicable to most biological mem- nonpolar amino acid residues of soluble
suggest that an analogy exists between branes, such as plasmalemmal and in- proteins, are no doubt of primary im-
the problems of the structure of mem- tracellular membranes, including the portance in determining the conforma-
branes and the structure of proteins. membranes of different cell organelles, tions that protein molecules adopt in
The latter are tremendously diverse in such as mitochondria and chloroplasts. aqueous solution (6), in which the non-
composition, function, and detailed These membranes are henceforth re- polar residues are predominantly se-
structure. Each kind of protein mole- ferred to as functional membranes. questered in the interior of the mole-
cule is structurally unique. Nevertheless,There may be some other membrane- cules away from contact with water.
generalizations about protein structure like systems, such as myelin, or the By hydrophilic interactions is meant a
have been very useful in understanding lipoprotein membranes of small animal set of thermodynamic factors that are
the properties and functions of protein viruses, which we suggest may be rigid, responsible for the preference of ionic
molecules. Similarly, valid generaliza- rather than fluid, mosaic structures, but and polar groups for an aqueous rather
tions may exist about the ways in whichsuch membrane systems are not a pri- than a nonpolar environment. For ex-
the proteins and lipids are organized in mary concern of this article. ample, the free energy required to trans-
an intact membrane. The ultimate test Our objectives are (i) to review brieflyfer a mole of zwitterionic glycine from
of such generalizations, or models, is some of the thermodynamics of macro- water to acetone is about 6.0 kcal at
whether they are useful to explain old molecular, and particularly membrane, 25?C, showing that ion pairs strongly
experiments and suggest new ones. systems in an aqueous environment; prefer to be in water than in a non-
Singer (1) has recently examined in (ii) to discuss some of the properties polar medium (1). These and related
of the proteins and lipids of functional free energy terms no doubt provide the
Dr. Singer is a professor of biology at the Uni-
versity of California at San Diego, La Jolla. Dr. membranes; (iii) to describe the fluid reasons why essentially all the ionic
Nicolson is a research associate at the Armand
Hammer Cancer Center of the Salk Institute for
mosaic model in detail; (iv) to analyze residues of protein molecules are ob-
Biological Studies, La Jolla, California. some of the recent and more direct served to be in contact with water,
720 SCIENCE, VOL. 175

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usually on the outer surface of the mol-
ecule, according to x-ray crystallo-
graphic studies. Similar thermodynamic
arguments apply to saccharide residues
(1). It requires the expenditure of sub-
Fig. 1. A phospho-spectrin (8) of erythrocyte membranes,
lipid bilayer: sche- which can be removed by chelating
matic cross-sectional
view. The filled cir-
agents under mild conditions, are ex-
cles represent the amples of membrane proteins that sat-
ionic and polar head isfy the criteria for peripheral proteins.

Ita?
stantial free energy to transfer a simple groups of the phos- On the other hand, the major portion
pholipid molecules, (> 70 percent) of the proteins of most
saccharide from water to a nonpolar which make contact
solvent, and such residues will therefore membranes have different characteris-
with water; the wavy
be in a lower free energy state in con- lines represent the tics, which may be assigned to integral
tact with water than in a less polar fatty acid chains. proteins: (i) they require much more
environment. drastic treatments, with reagents such
There are other noncovalent inter- as detergents, bile acids, protein dena-
actions, such as hydrogen bonding and (to the maximum extent feasible) from
turants, or organic solvents, to dissociate
electrostatic interactions, which also contact with water, while the ionic and them from membranes; (ii) in many in-
contribute to determine macromolecular polar groups of the proteins-along stances, they remain associated with
structure. However, with respect to gross with those of the lipids and the oligosac-lipids when isolated; (iii) if completely
structure, with which we are now charides-should be in contact with the freed of lipids, they are usually highly
concerned, these are very likely of sec- aqueous solvent. These requirements insoluble or aggregated in neutral aque-
ondary magnitude compared to hydro- place restrictions on models of mem- ous buffers (9).
phobic and hydrophilic interactions.brane structure; in particular, they ren- The distinction between peripheral
The familiar phospholipid bilayer der highly unlikely the classical model and integral proteins may be useful in
structure illustrates the combined effects of a trilaminar arrangement of a con- several regards. It is assumed that only
of hydrophobic and hydrophilic inter- tinuous lipid bilayer sandwiched be- the integral proteins are critical to the
actions. In this structure (Fig. 1) the tween two monolayers of protein. The structural integrity of membranes.
nonpolar fatty acid chains of the phos- latter model is thermodynamically un- Therefore, the properties and interac-
pholipids are sequestered together away stable because not only are the non- tions of peripheral proteins, while in-
from contact with water, thereby maxi- polar amino acid residues of the mem- teresting in their own right, may not be
mizing hydrophobic interactions. Fur- brane proteins in this model perforcedirectly relevant to the central prob-
thermore, the ionic and zwitterionic largely exposed to water but the ionic lems of membrane structure. The prop-
groups are in direct contact with the and polar groups of the lipid are se- erties of cytochrome c, for example,
aqueous phase at the exterior surfaces questered by a layer of protein frommay not be typical of mitochondrial
of the bilayer, thereby maximizing hy- contact with water. Therefore, neither membrane proteins. Furthermore, the
drophilic interactions. In the case of hydrophobic nor hydrophilic interac- biosynthesis of peripheral and integral
tions are maximized in the classical proteins and their attachment to the
zwitterionic phospholipids such as phos-
model. membrane may be very different proc-
phatidylcholine, dipole-dipole interac-
tions between ion pairs at the surface esses. This is not the appropriate oc-
of the bilayer may also contribute to casion to discuss membrane biogenesis
the stabilization of the ,bilayer structure. Some Properties of in any detail, but it may be significant
In applying these thermodynamic Membrane Components that, although cytochrome c is a mito-
principles to membranes, we recognize chondrial protein, it is synthesized on
first that of the three major classes of Peripheral and integral proteins. It cytoplasmic rather than mitochondrial
membrane components-proteins, lip- seems both reasonable and important ribosomes; in fact only a small fraction
ids, and oligosaccharides-the proteins to discriminate between two categories of the total mitochondrial protein (per-
are predominant. The ratio by weight of proteins bound to membranes, which haps only the integral proteins of the
of proteins to lipids ranges from about we have termed peripheral and integral inner mitochondrial membrane?) ap-
1.5 to 4 for those functional membranes proteins (1). Peripheral proteins may be pears to be synthesized on mitochon-
which have been well characterized characterized by the following criteria. drial ribosomes (10). In any event,
[compare (7)]. A substantial frac- (i) They require only mild treatments, because of the relatively unimportant
tion of this protein most probably playssuch as an increase in the ionic strength membrane structural role assigned to
an important role in determining theof the medium or the addition of a the peripheral proteins, they are not a
structure of membranes, and the struc-chelating agent, to dissociate themprimary mo- concern of this article.
tural properties of these proteins arelecularly intact from the membrane; Properties of integral proteins. Since
therefore of first-order importance. (ii) they dissociate free of lipids; and
the proteins we have classified as in-
Membrane proteins are considered in(iii) in the dissociated state they tegral, are according to the criteria speci-
some detail in the following section. At relatively soluble in neutral aqueous fied, constitute the major fraction of
this juncture, the significant point isbuffers. These criteria suggest that a
membrane proteins, we assume that the
that if hydrophobic and hydrophilic in- peripheral protein is held to the mem-
properties to be discussed apply to the
teractions are to be maximized and the brane only by rather weak noncovalent integral proteins.
lowest free energy state is to be at-(perhaps mainly electrostatic) interac- 1) For several well-characterized
tained for the intact membrane in an tions and is not strongly associated membrane systems, including erythro-
aqueous environment, the nonpolar with membrane lipid. The cytochrome cyte and other plasma membranes, and
amino acid residues of the proteins- c of mitochondrial membranes, which mitochondrial membranes, the proteins
along with the fatty acid chains of the can be dissociated free of lipids by high have been shown to be grossly hetero-
phospholipids-should be sequestered salt concentrations, and the protein geneous with respect to molecular
18 FEBRUARY 1972 721

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 weights (11). There is no convincing lar proteins are attached to the outer Two qualifications should be stressed,
evidence that there exists one predom- surfaces of a lipid bilayer (16) would however, concerning the bilayer form
inant type of membrane protein that is not be satisfactory because, among of membrane lipids. (i) None of the
specifically a structural protein; recent other reasons, it would require mem-evidence so far obtained for the bilayer
reports to the contrary have been with- brane thicknesses much larger than theform permits us to say whether the
drawn. We consider this heterogeneity 75 to 90 angstroms generally observed.bilayer is continuous or interrupted (1).
to be more significant for a general A model in which globular protein The calorimetrically observed phase
model of membrane structure than the molecules are intercalated within the transitions, for example, occur over a
fact that in a few specialized instances, membrane would, however, meet these broad temperature interval, allowing the
as in the case of disk membranes of restrictions. possibility that the cooperative unit in-
retinal rod outer segments (12, 13), Theaphospholipids of membranes. volved in the phase transition is quite
single protein species predominates. ThereAis now substantial evidence that small, consisting perhaps of only 100
satisfactory membrane model must the be
major portion of the phospholipidslipid molecules on the average. (ii) None
capable of explaining the heterogeneity is in bilayer form in a variety of intactof the experiments mentioned above is
of the integral membrane proteins. membranes. For example, differentialsufficiently sensitive and quantitative to
2) The proteins of a variety of calorimetry
intact of intact mycoplasma mem-prove whether 100 percent of the phos-
membranes, on the average, show ap-shows that they undergo a phase pholipid is in the bilayer form. It is
branes
preciable amounts of the a-helicaltransition
con- in a temperature range verytherefore not excluded that some signifi-
formation, as was first shown iby Keto that of aqueous dispersions ofcant fraction of the phospholipid (per-
similar
(14), Wallach and Zahler (4), and the phospholipids extracted from thehaps as much as 30 percent) is physi-
Lenard and Singer (3). For example, membranes (16, 17). Thus the structurescally in a different state from the rest
circular dichroism measurements of of the lipid in the membrane and of theof the lipid.
aqueous suspensions of intact and me-
lipid in isolated aqueous dispersion are Protein-lipid interactions in mem-
chanically fragmented human erythro- closely similar; presumably the latter is branes. Several kinds of experiments
cyte membranes (provided that we thetake
bilayer form. This conclusion is sup-indicate that protein-lipid interactions
into account certain optical anomalies ported iby x-ray diffraction '(18) andplay a direct role in a variety of
of these measurements) reveal that spir-label studies (19) on similar mem- membrane functions. Many membrane-
about 40 percent of the protein is inbrane preparations. bound enzymes and antigens require
the right-handed a-helical conformation The bilayer character of membranelipids, often specific phospholipids, for
(15). Most soluble globular proteins lipids rules out models such as that of the expression of their activities [see
whose circular dichroism spectra have Benson (20) in which the proteins and table 2 in (21)]. Furthermore, the
been obtained exhibit a smaller fraction lipids form a single-phase lipoproteinnature of the fatty acids incorporated
of a-helix in their native structures. subunit that is repeated indefinitely ininto phospholipids affects the function
two dimensions to constitute the mem-
This suggests that the integral proteins of certain membrane-bound proteins in
in intact membranes are largely globu- In such a model, most of the
brane. bacterial membranes (22).
lar in shape rather than spread out lipids
as would be expected to have dis- On the other hand, the calorimetric
monolayers. On the other hand, a tinctly different properties from thosedata discussed above give no significant
membrane model in which such globu- of a bilayer. indication that the association of pro-
teins with the phospholipids of intact
membranes affects the phase transitions
of the phospholipids themselves. Ex-
4. periments with phospholipase C and
membranes have shown that the en-
zymic release of 70 percent of the
phosphorylated amines from intact
erythrocyte membranes profoundly
perturbs the physical state of the resid-
ual fatty acid chains, but has no detect-
able effect (as measured by circular
dichroism spectra) on the average con-
formation of the membrane proteins
(2). Such results therefore suggest that
the phospholipids and proteins of
membranes do not interact strongly; in
Fig. 2. The lipid-globular protein fact, they appear to be largely
mosaic model inde- of m
cross-sectional view. The phospholipids pendent. are depicted as
a discontinuous bilayer with their ionic
This paradox, and
that different polar
types of head
lipid may be structurally differentiated from the bulk
experiments suggest strong protein-lipid
is not explicitly shown in the figure. The integral prote
senting the folded polypeptide chains, interactions on the are
one hand, and
shownweak as gl
bedded in, and partially protruding or no interactions
from, on the other, the
can be membr
on their surfaces the ionic residues (- and +) of the protein, while the nonpolar resolved in a manner consistent with
residues are largely in the embedded parts; accordingly, the protein molecules are am- all the data if it is proposed that, while
phipathic. The degree to which the integral proteins are embedded and, in particular,
whether they span the entire membrane thickness depend on the size and structure of the largest portion of the phospholipid
the molecules. The arrow marks the plane of cleavage to be expected in freeze-etchingis in bilayer form and not strongly
experiments (see text). [From Lenard and Singer (3) and Singer (1)] coupled to proteins in the membrane,
722 SCIENCE, VOL. 175

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a small fraction of the lipid is more adopt an amphipathic structure, protein
can be or in the association of two or
tightly coupled to protein. With any integral proteins of membranes; in this
more integral protein subunits to form
one membrane protein, the tightly manner, the heterogeneity of the a specific
pro- aggregate within the mem-
coupled lipid might be specific; that is, teins of most functional membranes can These features can be accom-
brane.
the interaction might require that the be rationalized. modated in Fig. 2 without any changes
phospholipid contain specific fatty acid The same considerations may also ex-in the basic structure.
chains or particular polar head groups. plain why some proteins are membrane- The phospholipids of the mosaic
There is at present, however, no satis- bound and others are freely soluble in structure are predominantly arranged as
factory direct evidence for such a dis- the cytoplasm. The difference may be an interrupted bilayer, with their ionic
tinctive lipid fraction. This problem is that either the amino acid sequence of and polar head groups in contact with
considered again in connection with a the particular protein allows it to adoptthe aqueous phase. As has been dis-
discussion of the experiments of Wilson an amphipathic structure or, on thecussed, however, a small portion of the
and Fox (23). contrary, to adopt a structure in whichlipid may be more intimately associated
the distribution of ionic groups is nearlywith the integral proteins. This feature
spherically symmetrical, in the lowest is not explicitly indicated in Fig. 2. The
Fluid Mosaic Model free energy state of the system. If thethickness of a mosaic membrane would
ionic distribution on the protein sur-vary along the surface from that across
Mosaic structure of the proteins and face were symmetrical, the proteina phospholipid bilayer region to that
lipids of membranes. The thermody- would be capable of interacting stronglyacross a protein region, with an average
namic considerations and experimental with water all over its exterior surface,value that could be expected to corre-
results so far discussed fit in with the that is, it would be a monodisperse sol-spond reasonably well to experimentally
idea of a mosaic structure for mem- uble protein. measured membrane thicknesses.
branes (1-3, 24) in which globular mol- The mosaic structure can be readily Matrix of the mosaic: lipid or pro-
ecules of the integral proteins (perhaps diversified in several ways. Although tein? In the cross section of the mosaic
in particular instances attached to oli-the nature of this diversification is a structure represented in Fig. 2, it is not
gosaccharides to form glycoproteins, matter of speculation, it is importantindicated
to whether it is the protein or the
recognize that the mosaic structure need
or interacting strongly with specific lip- phospholipid that provides the matrix of
ids to form lipoproteins) alternate with not be restricted by the schematic rep- the mosaic. In other words, which com-
sections of phospholipid bilayer in the resentation in Fig. 2. Protein-protein ponent is the mortar, which the bricks?
cross section of the membrane (Fig. 2). interactions that are not explicitly con- This question must be answered when
The globular protein molecules are pos- sidered in Fig. 2 may be important in the third dimension of the mosaic struc-
tulated to be amphipathic (3, 4) as are determining the properties of the mem- ture is specified. Trhese two types of
brane. Such interactions may result
the phospholipids. That is, they are mosaic structure may be expected to
either in the specific binding of have
structurally asymmetric, with one highly a very different structural and func-
peripheral protein to the exterior tional
polar end and one nonpolar end. The ex- properties, and the question is
posed surface of a particular integral
highly polar region is one in which the therefore a critical one. It is our hy-
ionic amino acid residues and any co-
valently bound saccharide residues are
clustered, and which is in contact with
the aqueous phase in the intact mem-
brane; the nonpolar region is devoid of
ionic and saccharide residues, contains
many of the nonpolar residues, and is
embedded in the hydrophobic interior
of the membrane. The amphipathic
structure adopted by a particular in-
tegral protein (or lipoprotein) molecule,
and therefore the extent to which it is
embedded in the membrane, are under
thermodynamic control; that is, they
are determined by the amino acid se-
quence and covalent structure of the
protein, and by its interactions with its
molecular environment, so that the free
energy of the system as a whole is at a
minimum. An integral protein molecule
with the appropriate size and structure,
or a suitable aggregate of integral pro-
teins (below) may transverse the entire
membrane (3); that is, they have re-
gions in contact with the aqueous sol- Fig. 3. The lipid-globular protein mosaic model with a lipid matrix (the fluid mosaic
vent on both sides of the membrane. model); schematic three-dimensional and cross-sectional views. The solid bodies with
It is clear from these considerations stippled surfaces represent the globular integral proteins, which at long range are
randomly distributed in the plane of the membrane. At short range, some may form
that different proteins, if they have thespecific aggregates, as shown. In cross section and in other details, the legend of
appropriate amino acid sequence toFig. 2 applies.
18 FEBRUARY 1972 723

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 area of the membrane surface. In able evidence. Some more recent re-
pothesis that functional cell membranes
have a long-range mosaic structure with sults, however, bear even more directly
other words, we suggest that long-range
the lipids constituting the matrix, as is random arrangements in membranes on are
the problem, and only this evidence
the norm; wherever nonrandom distri- is discussed below.
shown in Fig. 3. Supporting evidence is
discussed later. At this point, let us butions are found, mechanisms must
consider some of the consequences of exist which are responsible for them.
this hypothesis. 2) It has been shown that, under Some Recent Experimental Evidence
1) There should generally be no long- physiological conditions, the lipids of
range order in a mosaic membrane with functional cell membranes are in a Evidence for proteins embedded in
a lipid matrix. By long range, we mean fluid rather than a crystallinemembranes.
state. One proposal of the fluid
over distances of the order of a few mosaic model is that an integral pro-
(This is not true of myelin, however.)
tenths of a micrometer and greater. tein is
This evidence comes from a variety ofa globular molecule having a
Suppose we have a membrane prepara- significant fraction of its volume em-
sources, such as spin-labeling experi-
tion containing many different protein ments (25), x-ray diffraction bedded in the membrane. The results
studies
species, and suppose further that 10,000 (18), and differential calorimetry of recent
(16, freeze-etching experiments
with membranes strongly suggest that a
molecules of protein A are present in 17). If a membrane consisted of integral
the membrane of a single cell or or- substantial amount of protein is deeply
proteins dispersed in a fluid lipid matrix,
ganelle. How is protein A distributed the membrane would in effect be a two- embedded in many functional mem-
over the membrane surface? If the dimensional liquid-like solution of mon- branes. In this technique (26) a frozen
membrane proteins formed the matrix omeric or aggregated integral proteins specimen is fractured with a microtome
of the mosaic, defined by specific con- (or lipoproteins) dissolved in the lipid knife; some of the frozen water is sub-
tacts between the molecules of different bilayer. The mosaic structure would be limed (etched) from the fractured sur-
integral proteins, protein A might be a dynamic rather than a static one. Theface if desired; the surface is then
distributed in a highly ordered, two- integral proteins would be expected to shadow cast with metal, and the surface
dimensional array on the surface. On undergo translational diffusion within replica is examined in the electron mi-
the other hand, if lipid formed thethe membrane, at rates determined in croscope. By this method the topog-
matrix of the mosaic, there would be no part by the effective viscosity of the raphy of the cleaved surface is re-
long-range interactions intrinsic to the lipid, unless they were tied down by vealed. A characteristic feature of the
membrane influencing the distribution some specific interactions intrinsic orexposed surface of most functional
of A molecules, and they should there- extrinsic to the membrane. However,membranes examined by this technique,
fore be distributed in an aperiodic ran- because of their amphipathic structures,including plasmalemmal, vacuolar, nu-
dom arrangement on the membrane the integral proteins would maintain clear, chloroplast, mitochondrial, and
surface. their molecular orientation and their bacterial membranes (27, 28), is a
The absence of long-range order degree of intercalation in the membrane mosaic-like structure consisting of a
should not be taken to imply an ab- while undergoing translational diffusionsmooth matrix interrupted by a large
sence of short-range order in the mem- in the plane of the membrane (as dis- number of particles. These particles
brane. It is very likely that such short- cussed below). have a fairly characteristic uniform
range order does exist, as, for example, In contrast, if the matrix of the mo- size for a particular membrane, for
among at least some components of the saic were constituted of integral pro- example, about 85-A diameter for eryth-
electron transport chain in the mito- teins, the long-range structure of the rocyte membranes. Such surfaces re-
chondrial inner membrane. Such short- membrane would be essentially static. sult from the cleavage of a membrane
range order is probably mediated byLarge energies of activation would be along its interior hydrophobic face
required for a protein component to (29). This interior face (Fig. 2) corre-
specific protein (and perhaps protein-
diffuse in the plane of the membrane sponds to the plane indicated by the
lipid) interactions leading to the forma-
tion of stoichiometrically defined ag- from one region to a distant one be- arrow. If cleavage were to occur
gregates within the membrane. How- cause of the many noncovalent bonds smoothly between the two layers of
ever, in a mosaic membrane with a between the proteins that would have phospholipid in the bilayer regions, but
lipid matrix, the long-range distribu- to be simultaneously broken for ex- were to circumvent the protein mole-
tion of such aggregates would be ex- change to take place. Therefore, a cules penetrating the mid-plane of the
pected to be random over the entiremosaic membrane with a protein ma- membrane, then the alternating smooth
surface of the membrane. trix should make for a relatively rigid and particulate regions observed on the
The objection may immediately be structure with essentially no transla- freeze-etch surfaces can be readily ex-
raised that long-range order clearly tional diffusion of its protein compo- plained by a mosaic structure for the
exists in certain cases where differen- nents within the membrane. membrane (Fig. 2), provided that the
tiated structures (for example, synapses) From the discussion in this and the particles can be shown to be protein
are found within a membrane. We sug- previous section, it is clear that the in nature. That the particles are indeed
fluid mosaic model suggests a set of
gest, in such special cases, either that protein has been suggested by recent
short-range specific interactions amongstructural properties for functional experiments (30).
integral proteins result in the formationmembranes at least some of which can Another consequence of the mosaic
of an unusually large two-dimensionalbe tested experimentally. In an earliermodel, suggested from its inception
aggregate or that some agent extrinsic article (1), a large body of experimen-(3), is that certain integral proteins pos-
to the membrane (either inside or out- tal evidence was examined for its rele- sessing the appropriate size and struc-
side the cell) interacts multiply with vance to models of membrane structure. ture may span the entire thickness of
specific integral proteins to produce a It was concluded that a mosaic struc- the membrane and be exposed at both
clustering of those proteins in a limitedture was most consistent with the avail- membrane surfaces. Chemical evidence
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that a trans-membrane protein, whose the treated (sensitized) cells were lysed
Rho(D) antigen site (36) on the mem-
molecular weight is about 100,000, is at an air-water interface, so that the brane. Since the clusters were distrib-
present in large amounts in the human cell membranes were spread out flat. uted in a random array, we conclude
erythrocyte membrane has been ob- The flattened membranes, after being that the Rho(D) antigen, which exhibits
tained by two independent methods- picked up on an electron microscope properties of an integral protein (37),
one involving proteolysis of normal grid, were treated with the specific "in- is molecularly dispersed and is distrib-
compared to everted membranes (31), direct stain," ferritin-conjugated goat uted in a random ,two-dimensional array
and the other specific chemical labeling antibodies specific for human y-globu- on the human erythrocyte membrane.
of the membrane proteins !(32). lin. Thus, wherever the human anti-Rho Similar experiments were carried out
Distribution of components in the (D) molecules were bound to the Rho with the H-2 alloantigenic sites on
plane of the membrane. A prediction (D) antigen on the membrane surface, mouse erythrocyte membranes. In this
of the fluid mosaic model is that the the ferritin-labeled goat antibodies be- case (Fig. 5) the clusters -of ferritin
two-dimensional long-range distribution came specifically attached. In other molecules of the indirect stain were not
of any integral protein in the plane of words, the human y-globulin antibodyisolated, as in the case of the Rho(D)
the membrane is essentially random. now functioned as an antigen for the antigen, but instead occurred in patches.
To test this prediction, we have devel- goat antibodies (Fig. 4). The ferritin The patchy distribution of the H-2
oped and applied electron microscopic was distributed in discrete clusters, eachhistocompatibility alloantigenic sites had
techniques to visualize the distribution containing two to eight ferritin mole- earlier been observed by different tech-
of specific membrane antigens over clues within a circle of radius about niques (38), but the two-dimensional
large areas of their membrane surfaces 300 A. The numblers of such clusters distribution of the patches could not be
(33) and have so far studied the dis- per unit area of the membrane surface ascertained. In our experiments, the
tribution *of the Rhd(D) antigen on corresponded to the number of 125I- patches contained variable numbers of
human erythrocyte membranes (34), andlabeled human anti-Rho(D) molecules clusters, and were arranged in an ir-
of H-2 histocompatibility alloantigens bound per unit area. This indicates that regular two-dimensional array on the
on mouse erythrocyte membranes (35).each ferritin cluster was bound to a membrane surface. The histocompati-
In the case of the Rho(D) antigen, single anti-Rho(D) molecule, and a clus- bility antigen appears to be glycopro-
for example, cells of 0, Rh-positive ter represents the number of goat tein in nature (39). The long-range dis-
type were reacted with a saturating antibody molecules bound to a single trilbution of both the Rho(D) and H-2
amount of 125I-labeled purified humanhuman y-globulin molecule. Each clus-histocompatibility antigens on their re-
antibody to Rho(D) [anti-Rlho(D)], and ter therefore corresponds to a single spective membrane surfaces, therefore,

Fig. 4 (left). The outer membrane surface of an Rh-positive human erythrocyte sensitized with human anti-Rho(D) and stained with
ferritin-conjugated goat antibody to human y-globulin. The cells were first labeled to saturation with purified lIL-labeled human anti-
body to Rho(D) and then lysed at an air-water interface. The erythrocyte membrane ghosts, flattened by surface forces (inset,
low magnification) were picked up on a coated, electron microscope grid and indirectly stained with ferritin-conjugated goat anti-
bodies to human y-globulin. The ferritin appears bound to the membrane in discrete clusters of two to eight ferritin-conjugates; each
cluster is circumscribed by a circle of radius 300 A. The number of such clusters per cell (9300) is equal within experimental error to
the number of "2I-labeled human antibody to Rho(D) molecules bound per cell (10,200). Each cluster -therefore corresponds
to an individual Rho(D) antigenic site. Scale is 0.1 tum; inset scale is i ,tm. [From Nicolson, Masouredis, and Singer (34)] Fig.
5 (right). The outer membrane surface on a mouse erythrocyte (H-2b) sensitized with alloantibodies against H-2b histocompatibil-
ity antigens and stained with ferritin-conjugated antibodies against 7S mouse r-globulin. The procedures are the same as listed in
the legend to Fig. 4. The ferritin-antibody clusters are present in randomly spaced "patches" of variable size on the membrane sur-
face. Scale is 0.1 ,um. [From Nicolson, Hyman, and Singer (35)]

18 FEBRUARY 1972 725

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 are in accord with the prediction of the trated two-dimensional fluid solution of explained by a diffusion mechanism for
fluid mosaic model that the integral identical protein molecules will appear, the intermixing process, as follows. The
proteins of membranes are randomly when dried, to be arranged in an or- antibodies to the human cell membrane
arranged in two dimensions. dered array, particularly when optical were no doubt directed to a heteroge-
The particles on the inner membrane tricks are used to enhance the apparent neous set of antigens, whereas the anti-
faces revealed by freeze-etching experi- order (43). What is really a fluid phase bodies ito the mouse cell were directed
ments, which (as discussed above) are may therefore artifactually be made 'to specifically to the histocompatibility
probably protein in nature, are generally appear as a crystalline solid. This ap- alloantigen. However, the histocompati-
also relatively randomly distributed in pears to be the situation with the reti- bility antigens occur as large aggregates
two dimensions. nal receptor disk membranes. in the membrane (Fig. 5), and might
Evidence that proteins are in a A major contribution to membrane therefore be expected to diffuse more
fluid state in intact membranes. An im- studies has been made by Frye and slowly than a complex mixture of largely
portant series of experiments has been Edidin (44), who investigated the mem- unaggregated human antigens in the
carried out (12, 40-44) with receptor brane properties of some cell fusion membrane. Thus, at appropriate inter-
disk membranes from the retina of the heterokaryons. Human and mouse cells mediate times after cell fusion, signifi-
frog. This membrane system is unusual in culture were induced to fuse with cant numbers of (M1/2-HI) but not of
in that it contains as its predominant, one another, with Sendai virus as the (Ml-HI/2) fused cells might appear, to
if not only, protein component the pig- fusing agent. The distribution of humanbe converted at longer times to cells
ment rhodopsin. In electron microscopy and mouse antigenic components of the with completely intermixed components.
of the negatively stained surfaces of fused
the cell membranes was then deter- A rough estimate may be made of
dried membranes, a somewhat tightly mined by immunofluorescence, with thethe average effective diffusion constant
packed and ordered array of par- use of rabbit antibodies directed to the required of the membrane components
ticles (about 40 A) was observed. These whole human cells, mouse antibodies to account for the kinetics of intermix-
particles are the individual rhodopsin directed against the H-2 alloantigen on ing in the Frye-Edidin experiments.
molecules. Although the earlier studies the mouse cell membranes, and, as in- Taking the average distance of migra-
suggested that there was a long-range direct stains, goat antiserum to rabbit tion, x, to have been about 5 micro-
order in the distribution of the particles y-glolbulin and goat antiserum to mousemeters in a time, t, of 40 minutes gives
(40), more recent x-ray diffraction data y-globulin labeled with two different an apparent diffusion constant, D=x2/
(42) on pellets of wet, receptor disk fluorescent dyes. Shortly after cell fu- 2t, of 5 X 10-11 cm2/sec. For com-
membranes showed that only a few sion, the mouse and human antigenic parison, the diffusion constant of hemo-
orders of reflection were observed cor- components were largely segregated in glob,in in aqueous solutions is about
responding to the spacings of the rho- different halves of the fused cell mem- 7 X 10-7 cm2/sec. The apparent effec-
dopsin molecules in the plane of the branes; but after about 40 minutes at tive viscosity of the membrane fluid
membrane. This indicated that a non- 37?C the components were essentially phase is therefore about 103 to 104
crystalline, aperiodic arrangement ofcompletely intermixed. Inhibitors of times that of water.
the particles existed in the plane of theprotein synthesis, of adenosine triphos- The Frye-Edidin experiments can be
membrane. Furthermore, the tempera-phate (ATP) formation, and of gluta- rationalized by the fluid mosaic model
ture dependence of the characteristics mine-dependent synthetic pathways, ap- of membrane structure as being the re-
of the x-ray diffraction maxima wereplied before or after cell fusion, had no sult of the free diffusion and intermixing
consistent with the suggestion that the effect on the rate of this intermixing of the lipids and the proteins (or lipo-
particles were in a planar liquid-likeprocess, but lowering the temperature proteins) within the fluid lipid matrix.
state in the intact membrane. ,Additional below 15?C sharply decreased it. Some experiments, however, appear
support for the existence of this liquid- Frye and Edidin (44) suggest that to suggest that the lipids of membranes
like state was the observation that the the intermixing of membrane compo- are not readily interchangeable within
absorption of a foreign protein (bovinenents is due to diffusion of these com- the membrane and are therefore not
serum albumin) to the membrane couldponents within the membrane, rather free to diffuse independently. For ex-
definitely alter the x-ray spacings due than to their removal and reinsertion, ample, Wilson and Fox (23) have
to the rhodopsin particles; that is, the or to the synthesis and insertion of studied the induction of /l-galactoside
distribution of the rhodopsin moleculesnew copies of these components, into and 8/-glucoside transport systems in
in the plane of the membrane was rad-the heterokaryon membrane. An unex- mutants of Escherichia coli that cannot
ically altered by the weak binding ofplained finding of these experiments was synthesize unsaturated fatty acids. Such
the albumin. This alteration would not the fairly frequent occurrence, at early fatty acids can be lincorporated into
be expected if a rigid lattice structure and intermediate times after cell fusion, phospholipids, however, if they are sup-
of Ithe rhodopsin molecules, or aggre- of heterokaryon membranes in which plied in the growth medium. When cells
gates, were present in the plane of the the human antigenic components were were grown in particular fatty acid sup-
membrane, uniformly distributed over the mem- plements and induced for the synthesis
These studies are particularly note- brane surface but the mouse compo- of the transport systems, the effect of
worthy because they involved a mem- nents were still largely segregated to temperature on the transport rate was
brane which, by conventional electron about half the membrane (Ml/2-H1 characteristic of that fatty acid. If, then,
microscopic techniques, appears to showcells). On the other hand, the reverse the cells were first grown in medium
long-range periodicity over its surface. situation, with the mouse antigenic containing oleic acid and then shifted to
Other specialized membranes have also components uniformly spread out over growth in a medium supplemented with
exhibited ordered electron micrographic the membrane and the human compo- linoleic acid during a brief period of
images of their surfaces [compare (43)]. nents segregated (M1-H1/2), was only induction of either of the transport sys-
However, it is likely that a very concen- rarely observed. This result can now be tems, the effect of temperature on trans-
726
SCIENCE, VOL. 175

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port was said to be characteristic of cells charides on membranes in the electron surfaces must occur at only negligibly
grown continually in the linoleic acid microscope (33). For example, the fer-slow rates. This conclusion probably
medium. In other words, although most ritin conjugate of concanavalin A, aapplies to membrane proteins other
of the phopholipids of the membrane protein agglutinin that binds specificallythan glycoproteins; for example, the
contained oleic acid chains, these did to terminal a-D-glucopyranosyl or a-D- Na,K-dependent and Mg-dependent
not appear to exchange with the newly mannopyranosyl residues (46), attachesadenosine triphosphatase activities of
synthesized small amounts of phospholi- specifically to the outer surface of eryth-
erythrocyte membranes are exclusively
pids containing linoleic acid chains. rocyte membranes and not at all to localized to the inner cytoplasmic sur-
These experiments, however, do not the inner cytoplasmic surface (33). Afaces (51). Individual molecules of spin-
necessarily contradict the thesis that similar, completely asymmetric distri-labeled zwitterionic and anionic phos-
most of the phospholipids of membranes bution of ferritin conjugates of ricin (a pholipids also exhibit very slow inside-
are freely diffusible and, hence, ex- protein agglutinin) on the membranes outside transitions in synthetic vesicles
changeable. For example, each of the of rabbit erythrocytes is shown in Fig. of phospholipid bilayers (52). The very
two transport systems might be or- 6. Ricin binds specifically to terminalslow or negligible rates of such transi-
ganized in the membrane as a specific f/-D-galactopyranosyl and sterically re- tions can be explained by the mosaic
protein aggregate containing intercal- lated sugar residues (47). Such asym-model and the thermodynamic argu-
ated and strongly bound phospholipid metry has now been observed with ments already discussed. If the integral
components. If such lipoprotein aggre- several ferritin-conjugated agglutinins proteins (including the glycoproteins)
gates had first to be assembled in order and a number of different mammalian in intact membranes have, like the phos-
to be incorporated into the bulk lipid cell plasma membranes (48). These find- pholipids, an amphipathic structure, a
matrix of the membrane, the results of ings extend earlier results obtained by large free energy of activation would
Wilson and Fox would be anticipated. different methods (49). be required to rotate the ionic and polar
In particular, the small fraction of The foregoing observations bear on regions of the proteins through the
the membrane phospholipid that was many problems, including cell-cell inter-hydrophobic interior of the membrane
strongly bound, and perhaps segregated actions and membrane biogenesis (50).to the other side.
in such aggregates from the bulk of the In the context of this article, however, To accommodate the fluid mosaic
membrane lipid, might not exchange the absence of oligosaccharides on in- model to these conclusions concerning
rapidly with the bulk lipid. The Wilson- ner membrane surfaces indicates that asymmetry, we specify that, while the
Fox experiments therefore do not re- rotational transitions of the glycopro-two-dimensional translational diffusion
quire that the major part of the mem- teins of erythrocyte and other plasmaof the integral proteins and the phos-
brane phospholipid be static, but only membranes from the outer to the inner pholipids of membranes occurs freely,
that a small fraction of the lipids be
structurally differentiated from the rest.
The structural differentiation of some
of the membrane lipid by strong bind-
ing to integral proteins is a possibility
that was discussed above.
The observations of Wilson and Fox,
that there is a significant coupling of
lipid and protein incorporation into
membranes, appear to be a special case.
The experiments of Mindich (45) dem-
onstrate that more generally lipid and
protein incorporation into bacterial
membranes can occur independently,
and 'that quite wide variations in the ratio
of lipids and proteins in the membrane
can be produced in vivo, as might be
expected from the fluid mosaic model
of membrane structure.
The asymmetry of membranes. A
substantial amount of evidence has ac-
cumulated showing that the two sur-
faces of membranes are not identical
in composition or structure. One aspect
of this asymmetry is the distribution of
oligosaccharides on the two surfaces of
membranes. There exist plant proteins, Fig. 6. The inner (i) and outer (o) membrane surfaces of a rabbit erythrocyte mem-
brane that has been stained with ferritin-conjugated ricin. In preparing membrane speci-
called lectins or plant agglutinins, which
bind to specific sugar residues, and, as mens such as are shown in Figs. 4 and 5, occasionally a cell lyses with membrane
a result, can cause the agglutination of rupture such that both inner and outer surfaces of the membrane are exposed. In this
case the mounted membrane was stained with ferritin conjugated to ricin, a plant agglu-
cells bearing the sugar residues on their
tinin that specifically binds to terminal p-D-galactopyranosyl and sterically related
surfaces. By conjugating several such terminal sugar residues in oligosaccharides. The ferritin-agglutinin is found on the outer
membrane surface only. The scale is equivalent to 0.1 ,m; the insert scale is equiva-
agglutinins to ferritin, we have been able
lent to 1 Am.
to visualize the distribution of oligosac-
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the rotational diffusion of these com- sional solution, thereby allowing new ture, may be proposed. Consider first
ponents is generally restricted to axes thermodynamic interactions among the the proteolysis experiments with nor-
perpendicular to the plane of the mem- altered components to take effect. This mal cells. Suppose that the integral gly-
brane; that is, in general, molecular general mechanism may play an im- coproteins in the normal cell mem-
tumbling does not occur at significant portant role in various membrane-me- brane are molecularly dispersed in the
rates within the membrane. The asym- diated cellular phenomena that occur fluid mosaic structure. It is likely that
metry of the membrane introduces on a time scale of minutes or longer. mild proteolysis would preferentially
another factor into the problem of Much more rapidly occurring phenom- release a small amount of glycopeptides
translational diffusion of membrane ena, such as nerve impulse transmission, and other polar peptides from these
components. In the experiments of would find the mosaic structure to be a
Frye proteins because these are the most
and Edidin (44) only those membrane static one, insofar as translational diffu- exposed portions of the integral pro-
antigens exposed at the outer surface of sion of the membrane components is teins at the outer surface of the mem-
the membrane were labeled by fluores- concerned. In order to illustrate the brane (Figs. 2 and 3). The remaining
concepts involved, we discuss two spe-
cent antibodies, and the conclusion that portions of these proteins may still
cific membrane phenomena.
these particular antigens were mobile in contain a large fraction of their original
the plane of the membranes therefore, Malignant transformation of cells and oligosaccharide chains after the limited
strictly speaking, applies only to those the "exposure of cryptic sites." Normal proteolysis, but the release of some of
components accessible at the outer sur- mammalian cells grown in monolayer the more polar structures would make
face. Whether components confined to culture generally exhibit "contact in- the remaining portions more hydro-
the inner surfaces also intermix and hibition"; that is, they divide until they phobic. As these more hydrophobic
diffuse should be separately established. form a confluent monolayer and they glycoproteins diffused in the membrane,
Thus, recent evidence obtained with then stop dividing. Cells that have be- they might then aggregate in the plane
many experimental methods and differ- come transformed to malignancy by of the membrane. The result would be
ent kinds of functional membrane sys- oncogenic viruses or by chemical car- a clustering of the agglutinin-binding
tems is entirely consistent with the pre- cinogens lose the property of contact sites on the enzyme-treated cell sur-
dictions of the fluid mosaic model of inhibition; that is, they overgrow the face, as compared to the normal un-
membrane structure and provides strong monolayer. For some time, this experi- treated surface. Such clustering (with
support for the model. It seems amply mental finding has been thought to re- no increase, or perhaps even a decrease
justified, therefore, to speculate about flect the difference between the normal in the total numbers of sites because
how a fluid mosaic structure might and the malignant states in vivo, and of digestion) could enhance the agglu-
carry out various membrane functions, to be due to differences in the surface tination of the treated cells, as com-
and to suggest specific mechanisms for properties of normal and malignant pared to that of normal cells, because
various functions that can be subjected cells. Much excitement and investiga- it would increase the probability of
to experimental tests. tive activity therefore attended the dem-agglutinin bridges forming between the
onstration (53, 54) that malignant trans- surfaces of two cells.
formation is closely correlated with a In malignant transformation, distinct
The Fluid Mosaic Model and greatly increased capacity for the trans- chemical changes in the glycolipids and
Membrane Functions
formed cells to be agglutinated by sev- the glycoproteins of the cell membrane
eral saccharide-binding plant aggluti- are known to occur (56), and the en-
The hypothesis that a membrane is nins. Furthermore, mild treatment of hanced agglutinability of the transform-
an oriented, two-dimensional, viscous normal cells with proteolytic enzymes ed cells may be much more complicated
solution of amphipathic proteins (or can render them also more readily ag- than is the case in the proteolysis of
lipoproteins) and lipids in instantaneous glutinable by these protein agglutinins. normal cells. If, however, the two phe-
thermodynamic equilibrum, leads to Burger (54) has suggested, therefore, nomena do have a basic feature in com-
many specific predictions about the that the agglutinin-binding sites are pres-mon, it could be a similar clustering of
mechanisms of membrane functions. ent on the membrane surfaces of nor- saccharide-binding sites on the trans-
Rather than catalog a large number of formed and the enzyme-treated normal
mal cells but are "icryptic" (Fig. 7A)
these, we suggest some directions that (that is, they are shielded by some othercells. In malignant transformation, such
such speculations may usefully take. membrane components from effectively clustering could be the result of the
Among these problems are nerve im- participating in the agglutination proc- chemical changes in the membrane
pulse transmission, transport through ess), and that proteolytic digestion of mentioned above; or some virus-induced
membranes, and the effects of specific normal cells or the processes of malig- gene product (57) may be incorporated
drugs and hormones on membranes (1). nant transformation "exposes" these into the cell membrane and serve as a
The fluidity of the mosaic structure, cryptic sites on the membrane surface. nucleus for the aggregation of the ag-
which introduces a new factor into such In some cases, quantitative binding glutinin-binding glycoproteins within the
speculations, is emphasized here. This studies have indeed indicated that no membrane.
new factor may be stated in general significant change in the numbers of These suggestions can be tested ex-
form as follows. The physical or chem- agglutinin-binding sites on the mem- perimentally by the use of ferritin-con-
ical perturbation of a membrane may brane accompanies either mild pro- jugated agglutinins (33) as already dis-
affect or alter a particular membrane teolysis of normal cells or malignant cussed (Fig. 6). The prediction is that
component or set of components; a re- transformation (55). with normal cells subjected to mild
distribution of membrane components An alternative explanation of these proteolysis, and also with malignant
can then occur by translational diffu- phenomena (Fig. 7B), based on the transformed cells, the total number of
sion through the viscous two-dimen- fluid mosaic model of membrane struc- ferritin-agglutinin particles specifically
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bound to the outer surfaces of the cells and his co-workers (63) have pro- be quite different. The basic theory of
might not be greatly different from those posed an extension to membranes of Changeuex et al. (63) might still be
the Monod-Wyman-Changeux allosteric formally applicable, but with impor-
of normal cells, but larger clusters of
ferritin particles would be found. model of protein cooperative phenom- tant changes in physical significance. It
Cooperative phenomena in mem- ena, using as a model of membrane is possible, for example, that a particu-
branes. By a cooperative phenomenon structure an infinite two-dimensional lar integral protein can exist in either
we mean an effect which is initiated aggregate of identical lipoprotein sub- of two conformational states, one of
units [as, for example, the model de-which is favored by ligand binding; in
at one site on a complex structure and
transmitted to another remote site scribed
by by Benson (20)]. In this theo-its normal unbound conformation the
the treatment, the individual subunitsintegral protein is monomolecularly
some structural coupling betweenretical
two sites. A number of important are capable of existing in either of two dispersed within the membrane, but in
membrane phenomena may fall into conformational states, one of which the conformation promoted by ligand
this category. However, before enum- has a much larger binding affinity forbinding, its aggregation is thermody-
erating them, we should first discrimi- a specific ligand than does the other.namically favored. The binding of a
nate between two types of cooperative The binding of a single ligand moleculeligand molecule at one integral protein
effects that may occur. These can be to any one subunit then triggers thesite, followed by diffusion of the non-
termed trans and cis. Trans effects refer cooperative conversion of many of the liganded protein molecules to it, might
to cooperative (allosteric) changes that subunits to the ligand-bound confor-then lead to an aggregation and simul-
have been postulated to operate at some mation, in order to maximize the inter-taneous change in conformation of the
localized region on the membrane sur- actions among the subunits. aggregated protein within the mem-
face, to transmit an effect from one side This theory as presented relies on brane. This mechanism could result in
of the membrane to the other (58). For the membrane model used. If, however, a long-range cis-type cooperative phe-
example, fan integral protein may exist in the membrane is not a two-dimensional nomenon, if the eventual aggregate
the membrane as an aggregate of two (or aggregate of lipoprotein subunits, but size was very large and if its presence
more) subunits, one of which is expc,sed is instead a fluid mosaic of proteins produced local perturbations in the
to the aqueous solution at the outer sur- and lipids, the physical situation wouldproperties of the membrane. However,
face of the membrane, and the other
is exposed to the cytoplasm at the inner
surface. The specific binding of a drug
or hormone molecule to the active site
of the outward-oriented subunit may
induce a conformational rearrangement
within the aggregate, and thereby change
some functional property of the aggre- A
gate or of its inward-oriented subunit.
By cis effects, on the other hand, we
refer to cooperative changes that may
be produced over the entire membrane,
or at least large areas of it, as a conse-
quence of some event or events occur-
ring at only one or a few localized
points on the membrane surface. For
example, the killing effects of certain
bacteriocins on bacteria (59), the lysis
of the cortical granules of egg cells
upon fertilization of eggs by sperm
(60), and the interaction of growth
hormone with erythrocyte membranes
(61) are cases which may involve
B
transmission and amplification of local-
ized events over the entire surface of
a membrane. These phenomena may
not all occur by the same or related
mechanisms, but in at least two experi-
mental studies, that involving the inter-
w v
action of colicin E1 with intact Esche-
Fig. 7. Two different mechanisms to explain the findings that either malignantly
richia coli cells (62), and that of humanformed cells or normal cells that are subjected to mild proteolysis become
growth hormone and isolated humanmore readily agglutinable by several plant agglutinins. (A) The mechanism of
erythrocyte membranes (61), there is(54): agglutinin-binding sites that are present on the surfaces of normal cells,
substantial evidence that long-range cis-obstructed ("cryptic sites"), are exposed by proteolysis or the processes of ma
transformation. (B) The redistribution mechanism (see text): the agglutinin s
type cooperative effects intrinsic to the normal cell surfaces are largely monomolecularly dispersed in the fluid mosai
membranes are involved. ture, but on proteolysis or malignant transformation, they diffuse and aggr
The question we now address is, How clusters. The probability of agglutination of two such modified cells is enhan
might such cis effects work? Changeux the clustering of binding sites.
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 the transition would occur at a rate the membrane, an effect that may be ments with a wide variety of techniques
and over a time period determined by involved in the phenomenon of anti- and several different membrane sys-
the rate of diffusion of the molecules genic modulation (66). There are other tems are described, all of which are
of the integral protein in the fluid specific examples as well. consistent with, and add much detail
mosaic membrane. This time period is It may well be that a number of to, the fluid mosaic model. It therefore
likely to be relatively long, of the order critical metabolic functions performed seems appropriate to suggest possible
of minutes (44), as already mentioned. by cell membranes may require the mechanisms for various membrane
On the other hand, if cis-type cooper- translational mobility of some impor- functions and membrane-mediated
ative effects occurred in a lipoprotein tant integral proteins. This could be phenomena in the light of the model.
subunit model according to the mecha- the ultimate significance of the long- As examples, experimentally testable
nism postulated by Changeux et al. standing observation (67) that the mechanisms are suggested for cell sur-
(63), one would expect the coopera- membrane lipids of poikilothermic orga- face changes in malignant transforma-
tive change to be much faster. Con- nisms contain a larger fraction of un- tion, and for cooperative effects ex-
formation changes in the soluble allo- saturated fatty acids the lower their hibited in the interactions of membranes
steric protein aspartyltranscarbamylase, temperature of growth. Appropriate with some specific ligands.
for example, have half-times of the enzymes apparently carry out the nec- Note added in proof: Since this ar-
order of 10 milliseconds (64). It is essary biochemical adjustment (68) ticle was written, we have obtained
therefore of some interest that in the that keeps the membrane lipids in a electron microscopic evidence (69) that
studies of the interaction of colicin fluid state at the particular temperature the concanavalin A binding sites on
E1 and E. coli the fluorescence changes of growth; if these enzymes are not the membranes of SV40 virus-trans-
that marked the apparent cis-type co- functional, for example, because of formed mouse fibroblasts (3T3 cells)
operative transitions in the cell mem- mutations, the organism-to grow at are more clustered than the sites on the
brane occurred over intervals of one the lower temperature (65)-must be membranes of normal cells, as predicted
to several minutes (62). If this sug- supplied with the unsaturated fatty acid by the hypothesis represented in Fig.
gested mechanism for the colicin effect exogenously. While it has been sug- 7B. There has also appeared a study by
is valid, one would predict that (i) gested before that the maintenance of Taylor et al. (70) showing the re-
freeze-etching experiments on the coli- lipid fluidity may be important to markable effects produced on lympho-
cin-treated bacteria (28) might reveal carrier mechanisms operating across a cytes by the addition of antibodies di-
an aggregation of normally dispersed functional membrane, it is also possible rected to their surface immunoglobulin
particles at the inner membrane face, that the real purpose of fluidity is to molecules. The antibodies induce a re-
or (ii) changes in membrane fluidity, permit some critical integral proteins distribution and pinocytosis of these
such as would be produced by suitable to retain their translational mobility in surface immunoglobulins, so that within
changes in temperature or by different the plane of the membrane, as an about 30 minutes at 37?C the surface
compositions of membrane phospho- obligatory step in their function. immunoglobulins are completely swept
lipids (65), might markedly affect the out of the membrane. These effects do
kinetics of the fluorescence changes not occur, however, if the bivalent anti-
that are observed on addition of the Summary bodies are replaced by their univalent
colicin to the bacteria. Fab fragments or if the antibody ex-
In this discussion of membrane func- A fluid mosaic model is presented periments are carried out at 0?C in-
tions, some detailed mechanisms to for the gross organization and structure stead of 37?C. These and related results
account for two membrane phenomena of the proteins and lipids of biological strongly indicate that the bivalent anti-
have been presented. It may well turn membranes. The model is consistent bodies produce an aggregation of the
out that these mechanisms are incor- with the restrictions imposed by ther- surface immunoglobulin molecules in
rect. Our object has been not so much modynamics. In this model, the pro- the plane of the membrane, which can
to argue for these specific mechanisms, teins that are integral to the membrane occur only if the immunoglobulin mole-
as to illustrate that the fluid mosaic are a heterogeneous set of globular cules are free to diffuse in the mem-
model of membrane structure can sug- molecules, each arranged in an amphi- brane. This aggregation then appears to
gest novel ways of thinking about pathic structure, that is, with the ionic trigger off the pinocytosis of the mem-
membrane functions-ways that are and highly polar groups protruding brane components by some unknown
amenable to experimental tests. Other from the membrane into the aqueous mechanism. Such membrane transfor-
membrane phenomena may be influ- phase, and the nonpolar groups largely mations may be of crucial importance
enced by similar diffusional mechanisms: buried in the hydrophobic interior of in the induction of an antibody re-
for example, cell-cell and cell-sub- the membrane. These globular molecules sponse to an antigen, as well as in
strate interactions, where the apposition are partially embedded in a matrix of other processes of cell differentiation.
of intense local electric fields to a cell phospholipid. The bulk of the phosplho-
References and Notes
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5. W. Kauzmann, Advan. Protein Chem. 14, 1
lipid bilayer solvent. Recent experi-
tribution of the antigen in the plane of (1959).

730 SCIENCE, VOL. 175

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NEWS AND COMMENT study

study also
alsoindicates
indicatesthat
that
thethe
Soviet
Soviet
Union
Unionisisalmost
almostcertainly
certainly
pressing
pressing
ahead
ahead
-cautiously but intently-with a
manned lunar program that may be ex-
pected to put cosmonauts on the moon
The Soviet Space Program: Effort
as 1973. A related conclusion, perhaps
in the mid-1970's and possibly as early

Said to Surpass Peak U.S.study,

Level
the most surprising of the 670-page
is that the Russians may end up
spending the equivalent of $49 billion
to land men on the moon, far more
than the cost of the Apollo program.
A new and authoritative deaths
deaths
study of of
three
of cosmonauts
three cosmonauts
the last year. last or
Whether year.
not the Soviets actually
Soviet space program indicatesThe
The study,* that,
study,*
producedproduced
for the Senatefor the
carry Senate
through with their evident inten-
Committee
while American space efforts on Aeronautical and Space
continue tions, the study goes on, "it is not pos-
winding down toward the Sciences
lastby Apollo
analysts in three divisions sible to establish that the Russians have
of the
flight this year, the overall Library
Soviet of Congress, concludes invested smaller total resources in lunar
space
program remains "a strong that
andthe growing
current level of Soviet space exploration than the United States" even
activity exceeds by
enterprise," its ambitions unhindered that of the United though the Soviet effort "has not pro-
States at its
budgetary strain and undimmed bypeak
thein 1966. The space duced the visible result in this regard
which the United States has achieved."
* " Soviet Space Programs, 1966-70" Report of the Committee on Aeronautical and Space Sciences,
prepared by the Science Policy Research Division, Foreign Affairs Division, and the European LawThese and other findings stand in direct
Division, Library of Congress; available from the Government Printing Office, Washington, D.C.
20402, $3; stock number 5271-0263. contradiction of assertions by Soviet
18 FEBRUARY 1972 731

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