THE JOURNAL OF BIOLOGICAL CHEMISTRY

Vol. 275, No. 23, Issue of June 9, pp. 1722117224, 2000

2000 by The American Society for Biochemistry and Molecular Biology, Inc.
Printed in U.S.A.

Minireview
Structure and Function of
Sphingolipid- and
Cholesterol-rich
Membrane Rafts*
Published, JBC Papers in Press, April 18, 2000,
DOI 10.1074/jbc.R000005200
Deborah A. Brown and Erwin London
From the Departments of Biochemistry and Cell
Biology and Chemistry, State University of New York,
Stony Brook, New York 11794-5215
It is well known that separate domains with different lipid compositions can exist in liposomes containing mixtures of different
phospholipids. The question of whether cellular membranes contain
similar lipid domains has intrigued workers for many years. One type
of domain, sphingolipid and cholesterol-based structures called membrane rafts, has received much attention in the last few years. We
will review the evidence that rafts exist in cells and focus on their
structure, or the organization of raft lipids and proteins. Our discussion of function will focus on the role of rafts in signaling in hematopoietic cells, a particularly well developed area that has provided
insights into raft organization in the membrane. Several reviews of
rafts (1 4) and of related structures called caveolae (57) have appeared recently.
Lipid Phase Behavior and Raft Formation
Sphingolipids differ from most biological phospholipids in containing long, largely saturated acyl chains. This allows them to
readily pack tightly together, a property that gives sphingolipids
much higher melting temperatures (Tm)1 than membrane (glycero)phospholipids, which are rich in kinked unsaturated acyl
chains. It is now clear that tight acyl chain packing is a key feature
of raft lipid organization (3, 8, 9). In fact, the differential packing
ability of sphingolipids and phospholipids probably leads to phase
separation in the membrane. Thus, sphingolipid-rich rafts co-exist
with phospholipid-rich domains that are in the familiar, loosely
packed disordered state (variously abbreviated as L, lc, or ld).
Phase separation between lipids in different physical states, most
often the lc and the solid-like gel phases, has been well characterized in model membranes. Indeed, the gel phase is the most familiar state in which acyl chains are highly ordered.
However, because of the high concentration of cholesterol in the
plasma membrane and other membranes in which rafts form, raft
lipids do not exist in the gel phase. Cholesterol has important
effects on phase behavior. It is well known that addition of cholesterol to a pure phospholipid bilayer abolishes the normal sharp
thermal transition between gel and lc phases, giving the membrane
properties intermediate between the two phases. This effect initially suggested that domains in ordered and disordered states
cannot co-exist at high cholesterol levels. However, further work
showed that a different kind of phase separation can occur in
binary mixtures of individual phospholipids with cholesterol. In
these mixtures, domains in an lc-like phase co-exist with domains
in a new state, the liquid-ordered (lo) phase. Acyl chains of lipids in
* This minireview will be reprinted in the 2000 Minireview Compendium,
which will be available in December, 2000. This work was supported by
National Institutes of Health Grants GM 47897 (to D. A. B.) and GM 48596
(to E. L.).
To whom correspondence should be addressed. Tel.: 631-632-8563; Email: deborah.brown@sunysb.edu.
1
The abbreviations used are: Tm, melting temperature; lc, liquid crystalline; ld, liquid disordered; lo, liquid ordered; PE, phosphatidylethanolamine;
PC, phosphatidylcholine; DRM, detergent-resistant membrane; GPI, glycosylphosphatidylinositol; POPC, palmitoyl oleoyl PC; TCR, T cell receptor.
This paper is available on line at http://www.jbc.org

the lo phase are extended and tightly packed, as in the gel phase,
but have a high degree of lateral mobility (3).
Rafts probably exist in the lo phase or a state with similar properties. In support of this model, detergent-insoluble membranes that
can be isolated from cell lysates and are likely to be derived from rafts
(discussed below) are in the lo phase (10, 11). Model membrane
studies that do not involve detergents also support the idea that lo
phase and lc phase domains could co-exist in biological membranes.
These studies showed that phase separation can occur in ternary
mixtures of cholesterol with two phospholipids (or a phospholipid and
a sphingolipid) that have different Tm and thus different tendencies
to form an ordered phase (8, 12). In these mixtures, lo phase domains
enriched in the high Tm lipid separate from lc phase domains enriched in the low Tm lipid. Because of the significant difference in Tm
between sphingolipids and biological phospholipids, these lipid mixtures are a reasonable (though crude) model of cholesterol-containing
cell membranes like the plasma membrane.
Cholesterol has another important effect on phase behavior. As
discussed above, there are parallels between lo/lc phase separation
and gel/lc phase separation. In both cases, a phase in which acyl
chains are highly ordered (gel or lo) separates from a phase in
which they are disordered (lc). Thus, lipid mixtures can undergo
either gel/lc phase separation in the absence of cholesterol or lo/lc
phase separation in its presence. Comparing the phase behavior of
mixtures with and without cholesterol shows that the sterol can
sometimes promote phase separation (8, 12), apparently because of
favorable packing interactions between saturated lipids and sterol
(13). Thus, in phospholipid/sphingolipid mixtures, less sphingolipid
is required to form the lo phase (in the presence of cholesterol) than
to form the gel phase in its absence (8, 9). This cholesterol effect
probably explains why rafts can form in cell membranes that contain relatively low levels of sphingolipids. It also explains why
cholesterol depletion can disrupt rafts and affect raft function.
Finally, it probably explains why sphingomyelin, with a Tm of
37 41 C, can be essentially as effective as glycosphingolipids
(which can have much higher Tm) in promoting raft formation (11).
Any difference in raft stability that might result from the difference
in Tm between the two lipids is minor compared with the strong
raft-stabilizing effect of cholesterol.
Because headgroup structure is an important modulator of lipid
packing, headgroup as well as acyl chain structure may be important in raft formation. For instance, phosphatidylethanolamines
(PE), with their small headgroup, have much higher Tm than the
corresponding phosphatidylcholines (PC). This effect may be especially important in the sphingolipid-poor (but PE-rich) inner bilayer leaflet, where raft structure is very poorly understood.
Rafts and Detergent-insoluble Membranes
Membrane fragments that are insoluble in non-ionic detergents
(DRMs; also termed DIGs (detergent-insoluble glycolipid-enriched
membranes), GEMs (glycolipid-enriched membranes), and TIFF
(Triton-insoluble floating fraction)) can be isolated from most mammalian cells (14). DRMs appear to be derived from rafts; they are
rich in cholesterol and sphingolipids and are in the lo phase when
isolated from cells (10). Furthermore, lo phase liposomes are also
detergent-insoluble under the conditions used to extract cells (9).
Thus, there is a close relation between rafts and DRMs, and isolation of DRMs is one of the most widely used methods for studying
rafts.
The tight acyl chain packing of both gel and lo phase lipids is
probably responsible for their detergent insolubility. This provides
a rational explanation for the detergent insolubility of DRMs,
which was initially puzzling; in a tightly packed state, lipid-lipid
interactions can be more stable than lipid-detergent interactions.
DRM ProteinsA number of proteins are enriched in DRMs.
Some of these are targeted to rafts by modification with saturated
chain lipid groups, which pack well into an ordered lipid environ-

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Minireview: Membrane Rafts

ment. These modifications include glycosylphosphatidylinositol

(GPI) anchors and closely spaced myristate and palmitate or dual
palmitate chains (2, 1518). In contrast, both membrane-spanning
proteins and prenyl groups (which are bulky and branched) should
be difficult to accommodate in a highly ordered environment. Indeed, DRMs are relatively poor in transmembrane proteins and
contain very low levels of prenylated proteins (17).
Nevertheless, several specific transmembrane proteins are enriched in DRMs. Very little is known about how this occurs. Palmitoylation can contribute to DRM targeting (16, 17), although not all
palmitoylated transmembrane proteins are in DRMs and not all
transmembrane DRM proteins are palmitoylated. As might be
expected, the sequence of the membrane-spanning domain (which
could affect the way the protein interacts with lipids) can affect
DRM localization (19 21). However, mutations in cytoplasmic domains, which seem unlikely to interact directly with lipids, can also
affect DRM association (2225). Although the mechanism of this
effect is not known, such mutants might fail to interact with binding partners that themselves associate directly with raft lipids or
might be mistargeted to membranes whose lipid composition cannot support raft formation.
Clustering and DRM AffinityThe affinity of gangliosides (26)
and lipid-linked proteins (15, 27) for DRMs (and presumably also
for rafts) can be increased by clustering or oligomerization because
of the increase in the number of saturated acyl chains per molecule
or cluster. Enhancement of raft affinity by clustering of molecules
that individually have more modest raft affinity is supported by
theoretical considerations and may have important physiological
consequences (2, 27). This effect probably explains why several
receptors on the surface of hematopoietic cells are recruited to
DRMs when they are clustered following antigen binding (discussed below), although the structural features of these proteins
that confer an affinity for rafts have not been identified.
Limitations of the DRM MethodAlthough DRM association is a
useful way of showing that a protein or lipid has an affinity for
rafts, it cannot be used to quantitate the fraction of the molecule
that is present in rafts in the intact cell. This is partly because cells
must generally be chilled before detergent extraction in order to
isolate DRMs. Chilling is necessary to stabilize the lo phase and
enhance its detergent resistance. However, because phase separation is also strongly temperature-dependent, more of the membrane is probably in the lo phase at 0 than at 37 C. This may
explain why a surprisingly high fraction of plasma membrane
lipids can be detergent-insoluble (reviewed in Refs. 2 and 3) and
why the phospholipid composition of DRMs can be similar to that of
the plasma membrane (28). On the other hand, in some cases
detergent may partially solubilize raft lipids and proteins even
after chilling, leading to an underestimation of the fraction of these
molecules in rafts (11, 15, 29).
These temperature effects raise the question of whether rafts exist
at all in cell membranes at physiological temperatures and highlight
the importance of detergent-independent methods (described below)
in providing evidence for the existence of rafts.
Other Features of Raft Structure:
Outstanding Questions
Visualization of RaftsIf rafts are present in cells, it should be
possible to detect them by microscopy. Nevertheless, molecules
such as GPI-anchored proteins and gangliosides, taken as putative
raft markers because of their enrichment in DRMs, often appear
uniformly distributed on the cell surface when detected microscopically (reviewed in Refs. 3 and 4). However, the distribution of these
markers can change dramatically upon clustering with antibodies
or other agents. In some cases, clustering of one marker can cause
redistribution of another, although the two are unlikely to interact
directly (2, 27, 30 32) (discussed below in the section on hematopoietic cell signaling). The implication that the two markers colocalize because both are associated with rafts provides some of the
strongest evidence to date that rafts are present in cell membranes.
Why are rafts difficult to see without clustering? At least three
explanations are possible. First, rafts may be too small to see by
microscopy. If so, clustering might cause coalescence of rafts into
larger, visible units. Second, if individual markers have only a
moderate affinity for rafts, they may not be highly concentrated in

FIG. 1. Model of raft organization. Tightly packed sphingolipids are

enriched in lo phase rafts (light gray) in the cholesterol-rich plasma membrane. Clustering of a protein that has an affinity for rafts (indicated by
arrows) could either cause small, dispersed rafts containing the protein to
coalesce into larger rafts (A), or increase the overall raft affinity of the
protein cluster enough to recruit it to rafts (B). Clustering of proteins could
even induce raft formation (3, 27).

the domains. For instance, rafts would be difficult to detect visually

if the concentration of a marker there were only 3- or 4-fold higher
than in the rest of the membrane. These values are plausible
because phase diagrams indicate that saturated acyl chains can
sometimes give lipids only a moderate preference for ordered membrane domains (33). As discussed earlier, clustering could increase
the affinity of these markers for rafts, increasing their concentration in the domains and facilitating detection. Finally, it is even
possible that rafts do not exist constitutively but that clustering of
components that have an affinity for an ordered state induces raft
formation. We have discussed these alternate models of raft structure and dynamics previously (3), and they are illustrated in Fig. 1.
Raft SizeAs is clear from the previous discussion, raft size is
very poorly understood. Microscopically detectable lipid domains
can sometimes be observed in model membranes without cholesterol (34, 35), but it is not yet clear how cholesterol affects domain
size. Single-particle tracking experiments in cells showed that a
ganglioside and a GPI-anchored protein were transiently confined
to domains of about 200 300 nm that were sensitive to a glycosphingolipid synthesis inhibitor (4). Two other groups have used
resonance energy transfer to search for clustering of GPI-anchored
proteins in the membrane (36, 37). One group (36) found cholesteroldependent clustering into domains of 70 nm, too small to see by
microscopy, whereas the other found no evidence for clustering
(37). This discrepancy would be reconciled if in fact only a small
fraction of the molecules were closely clustered, a possibility that is
consistent with both studies (37). It should be noted, however, that
preferential partitioning of proteins into rafts does not necessarily
result in their close clustering. For instance, GPI-anchored proteins could be present at a relatively low concentration in rafts if
they had only a moderate affinity for the domains. Furthermore, as
there is no reason to suspect that GPI-anchored proteins in rafts
interact with each other more strongly than with raft lipids, they
may be uniformly distributed within rafts. Thus, GPI-anchored
proteins could be present at low local density if a large fraction of
the membrane formed rafts.
Membrane DistributionRafts are likely to be most abundant in
membranes that are rich in cholesterol and sphingolipids, including the plasma membrane, late secretory pathway, and endocytic
compartments. In the plasma membrane, rafts appear to have a

Minireview: Membrane Rafts

preferential association with 50 100-nm pits called caveolae,
which are present in many mammalian cell types (57). It should
be noted, though, that rafts (as detected by DRM formation and by
the co-clustering of several putative raft components) are not restricted to caveolae and are abundant in cells that lack caveolae. It
is not yet known whether localization in caveolae affects the structure of rafts.
The behavior of rafts in the cytoplasmic leaflet of the bilayer is
an important outstanding question. Although sphingolipids are
largely restricted to the outer leaflet, several observations suggest
that rafts are present in the inner leaflet and that rafts in the two
leaflets are coupled. First, DRMs have a bilayer appearance when
isolated and are enriched in dually acylated cytofacial membrane
proteins, such as Src family kinases and G protein subunits.
Second, Src family kinases can co-redistribute when cell-surface
GPI-anchored proteins or gangliosides are clustered using antibodies or toxins (27, 31).
Recent advances in tandem electrospray mass spectrometry
have shown that plasma membrane phospholipids are more highly
saturated than those in intracellular membranes (28, 38) and thus
may form rafts readily in the presence of cholesterol. The monounsaturated phospholipid palmitoyl oleoyl phosphatidylcholine
(POPC) may form the lo phase when mixed with cholesterol (39). In
addition, mixtures of brain PC (which is rich in POPC) and cholesterol are partially Triton-insoluble (in the cold) in the absence of
sphingolipids (9). Finally, as mentioned earlier, the high concentration of relatively high Tm PE in the inner leaflet may promote
raft formation. Together, these observations suggest that formation of lo phase rafts in the sphingolipid-poor cytoplasmic membrane leaflet is plausible.
How these rafts might be coupled with outer leaflet rafts is very
poorly understood. There is some evidence for monolayer coupling
of sphingolipid-rich domains in model membranes (40). More recently, confocal imaging and fluorescence correlation spectroscopy
were used to show a correlation between like phases in the two
leaflets of model membranes containing two phospholipid species
(34). Further characterization of the basis of this effect will probably shed light on the behavior of rafts in cells.
Introduction to Raft Function
In principle, targeting of proteins to rafts might affect function in
either of two ways. First, concentration of proteins in rafts could
facilitate interactions between them. (Similarly, segregation of raft
and non-raft proteins could separate them during sorting.) Second,
the ordered lipid environment might directly affect function, possibly by altering protein conformation. There are no clear examples
of this second possibility, although cholesterol concentration and
bilayer width can affect transmembrane helix orientation and helix-helix interaction (41, 42). In addition, cholesterol depletion
(which can disrupt raft function) alters the function of a raftassociated potassium channel (43).
Several approaches have been taken to investigate raft function.
One is to show that a protein is enriched in DRMs. This is consistent with a role for rafts in the function of that protein but does not
prove it. More suggestive is showing that several proteins that
must interact to function all redistribute and colocalize with each
other when one raft component is experimentally clustered. Another approach is to show that disrupting the association of a
protein with rafts disrupts function. For example, mutation of
palmitoylation sites on two proteins (Lck and LAT, described below) simultaneously abolished DRM association and affected function. Finally, raft disruption by depletion of cholesterol (or occasionally sphingolipids) can affect function. Although this method is
useful, cholesterol depletion may have pleiotropic effects on membrane structure and lipid-protein interactions in addition to disrupting rafts. The strongest evidence for the involvement of rafts in
function is provided when several approaches point to the same
conclusion. This is well illustrated in the case of signaling in
hematopoietic cells, particularly T cells and basophils.
Before examining this topic in detail, we will briefly mention
other processes in which rafts have been implicated. Rafts were
first proposed to mediate sorting in the trans-Golgi network, especially in polarized epithelial cells and neurons (1, 2, 44 46). Recent
results suggest that rafts may also be important in sorting in the

17223

endocytic pathway (47). Rafts can serve as docking sites for certain
pathogens and toxins (48). In addition, they may be important in
the aberrant amyloid precursor protein processing that contributes
to Alzheimers disease (6, 7). Integrin receptors may also function
in rafts. Several integrins have been found in DRMs (2, 7, 49, 50).
In one study, integrins, the integrin-associated protein IAP (which
can regulate integrin function), and heterotrimeric G proteins
formed a stable cholesterol-dependent complex that was enriched
in DRMs (49). Finally, rafts polarize to the front of adenocarcinoma
cells migrating in a chemotactic gradient (51). In these cells, development of front-rear polarity, which is required for directed
migration, is abolished by cholesterol depletion.
Signaling in Hematopoietic Cells
Although different hematopoietic cells play distinct roles in the
immune response, their antigen-responsive signaling pathways
have several features in common with each other. Multisubunit
receptors on T and B lymphocytes bind antigen directly, whereas
those on other hematopoietic cells constitutively bind the Fc domains of different classes of antibodies and thus bind antigen
indirectly (52). As examples of the latter case, FcR on neutrophils
binds IgG, FcRI on basophils and mast cells binds IgE, and FcR
on several myeloid cells binds IgA. In each case, antigen binding
triggers receptor cross-linking, leading to activation of specific Src
family tyrosine kinases. The activated kinases phosphorylate tyrosine residues in the cytoplasmic domains of one or more receptor
subunits. These events initiate signaling cascades, via recruitment
of downstream signaling proteins, that culminate in cell-type specific responses. As a specific example, an early signaling event in T
cells is the heavy tyrosine phosphorylation of the linker for activation of T cells (LAT) protein. Phosphorylated tyrosine residues on
LAT serve as the docking site for a number of downstream signaling proteins (16).
In several cases, receptors appear uniformly distributed on resting cells but can be experimentally clustered into large patches
using specific antibodies in a process that mimics physiological
clustering of receptors by antigen. As discussed later, this receptor
clustering can induce clustering of rafts.
DRM/Raft LocalizationSeveral of the signaling proteins described above are enriched in DRMs. Src family kinases and LAT
(16), both of which require acylation for DRM targeting, are present
in DRMs constitutively. Antibody-mediated clustering can recruit
receptors on several cell types to DRMs. These include the T cell
receptor (TCR) (53), the B cell receptor (54), FcRI (29), FcR (55),
and CD20 (23) (a protein whose cross-linking activates B cells).
There is some information on structural features of these transmembrane receptors that is required for their targeting to DRMs
(21, 23, 55), although no general patterns have emerged.
In several cases, receptor clustering induces redistribution of
other putative raft markers. As this co-clustering involves two sets
of molecules that are not believed to interact directly, it suggests
that both molecules associate with the same rafts and that these
coalesce into larger domains upon clustering of one component. As
an example, the ganglioside GM1 and other order-preferring lipids,
taken as raft markers, colocalize with clustered FcRI on basophils
(2). (It should be noted that GM1 is generally detected using cholera toxin. As this toxin is pentavalent, it should induce GM1
clustering, enhancing raft association.) In another example, the Src
family kinase Lyn, a signaling partner of FcRI in basophils, colocalizes with clustered FcRI in a cholesterol-dependent manner
(56). Similarly, contact of T cells with beads coated with antibodies
to the CD3 component of the TCR and to the co-stimulatory protein
CD28 induces clustering of GM1 at the contact site (30). Clustering
of gangliosides on T cells was also found to induce co-clustering of
the TCR, LAT, and the Src family kinase Lck (31, 57).
Functional Importance of Raft LocalizationSeveral observations support a functional role for rafts in hematopoietic cell signaling (reviewed in Refs. 58 and 59). FcRI becomes tyrosinephosphorylated by Lyn with the same kinetics with which it
becomes recruited to DRMs, and the fraction of the receptor that
partitions into DRMs is selectively phosphorylated (29). Cholesterol depletion inhibits receptor tyrosine phosphorylation in T cells
(60) and basophils (56) and signaling in T cells (60). Recruitment of
CD48 (a raft-associated GPI-anchored protein) to the site of contact

17224

Minireview: Membrane Rafts

between the TCR and antigen-presenting cell (APC) can enhance

signaling in a cholesterol-dependent manner (61). Another study
showed that patching of gangliosides in T cells stimulates signaling
(31).
Mutagenesis has been used to suggest that two proteins, Lck and
LAT, must be in rafts to function. Stimulation of signaling in T cells
by ganglioside patching (31) was abolished in cells expressing only
a mutant, non-palmitoylated form of Lck that cannot associate with
rafts. Strikingly, signaling was rescued when separate clusters of
the mutant Lck and of gangliosides were brought together using
bridging antibodies, demonstrating that the Lck was not inactivated by mutation. These results suggest that Lck must be physically close to its signaling partners for productive signaling and
that this association normally requires clustering of the proteins in
rafts.
A similar approach suggests that LAT must be in rafts to function (62). A mutant non-palmitoylated LAT is localized correctly to
the plasma membrane but is absent from DRMs. This protein
cannot serve as a substrate for tyrosine phosphorylation (62) or
function in signaling (63).
Other studies suggest further roles for rafts in hematopoietic cell
function. For instance, signaling through a transmembranous
FcR on human neutrophils is enhanced if the receptor is coclustered with any of several GPI-anchored proteins, including an
endogenous GPI-anchored form of FcR (64). As GPI-anchored
proteins are enriched in DRMs, this result suggests that signaling
is enhanced through recruitment of rafts to clusters of transmembrane FcR. In further support of that conclusion, when the GPIanchored FcR was clustered using antibodies specific for that form
of the protein, the transmembrane FcR associated with the clusters (65). Another study showed that GM1 (taken as a raft marker)
polarizes to the site of contact between natural killer lymphocytes
and their target cells in a cholesterol-dependent manner (66). Finally, adhesion of T lymphocytes to other cells via binding of the
integrin LFA1 to intercellular adhesion molecules can be stimulated by clustering of gangliosides or of a GPI-anchored protein on
the T cells, also in a cholesterol-dependent manner (50).
Signaling in Other CellsRafts may play a role in signaling
outside hematopoietic cells, although this area has not yet been
well developed. One example involves ephrin B proteins, which are
important in the developing nervous system. Ephrin B proteins,
which are in DRMs, were recently shown to recruit multiprotein
signaling complexes to DRMs when expressed exogenously (24).
Other studies using fibroblasts showed that cholesterol depletion
inhibits hormone-stimulated phosphatidylinositol turnover, probably by delocalizing polyphosphoinositide 4,5- bisphosphate from
rafts (67) and causes hyperactivation of extracellular signal-regulated kinase in response to epidermal growth factor (68). Finally, a
number of studies suggest that caveolae are important signaling
centers (5, 7).
AcknowledgmentWe thank Anne Kenworthy for sharing data before
publication.
REFERENCES
1.
2.
3.
4.
5.
6.
7.

8.
9.
10.
11.
12.
13.

Simons, K., and Ikonen, E. (1997) Nature 387, 569 572

Brown, D. A., and London, E. (1998) Annu. Rev. Cell Dev. Biol. 14, 111136
Brown, D. A., and London, E. (1998) J. Membr. Biol. 164, 103114
Jacobson, K., and Dietrich, C. (1999) Trends Cell Biol. 9, 8791
Anderson, R. G. W. (1998) Annu. Rev. Biochem. 67, 199 225
Kurzchalia, T. V., and Parton, R. G. (1999) Curr. Opin. Cell Biol. 11, 424 431
Smart, E. J., Graf, G. A., McNiven, M. A., Sessa, W. C., Engelman, J. A.,
Scherer, P. E., Okamoto, T., and Lisanti, M. P. (1999) Mol. Cell. Biol. 19,
7289 7304
Ahmed, S. N., Brown, D. A., and London, E. (1997) Biochemistry 36,
10944 10953
Schroeder, R. J., Ahmed, S. N., Zhu, Y., London, E., and Brown, D. A. (1998)
J. Biol. Chem. 273, 1150 1157
Ge, M., Field, K. A., Aneja, R., Holowka, D., Baird, B., and Freed, J. (1999)
Biophys. J. 77, 925933
Ostermeyer, A. G., Beckrich, B. T., Ivarson, K. A., Grove, K. E., and Brown,
D. A. (1999) J. Biol. Chem. 274, 34459 34466
Silvius, J. R., del Guidice, D., and Lafleur, M. (1996) Biochemistry 35,
15198 15208
Xu, X., and London, E. (2000) Biochemistry 39, 844 849

14. Brown, D. A., and Rose, J. K. (1992) Cell 68, 533544

15. Arni, S., Keilbaugh, S. A., Ostermeyer, A. G., and Brown, D. A. (1998) J. Biol.
Chem. 273, 28478 28485
16. Zhang, W., Trible, R. P., and Samelson, L. E. (1998) Immunity 9, 239 246
17. Melkonian, K. A., Ostermeyer, A. G., Chen, J. Z., Roth, M. G., and Brown, D. A.
(1999) J. Biol. Chem. 274, 3910 3917
18. Moffett, S., Brown, D. A., and Linder, M. E. (2000) J. Biol. Chem. 275,
21912198
19. Perschl, A., Lesley, J., English, N., Hyman, R., and Trowbridge, I. S. (1995)
J. Cell Sci. 108, 10331041
20. Scheiffele, P., Roth, M. G., and Simons, K. (1997) EMBO J. 16, 55015508
21. Field, K. A., Holowka, D., and Baird, B. (1999) J. Biol. Chem. 274, 17531758
22. Puertollano, R., and Alonso, M. A. (1998) J. Biol. Chem. 273, 12740 12745
23. Polyak, M. J., Tailor, S. H., and Deans, J. P. (1998) J. Immunol. 161,
32423248
24. Bruckner, K., Labrador, J., Scheiffele, P., Herb, A., Seeburg, P., and Klein, R.
(1999) Neuron 22, 511524
25. Machleidt, T., Li, W.-P., Liu, P., and Anderson, R. G. W. (2000) J. Cell Biol.
148, 1728
26. Hagmann, J., and Fishman, P. H. (1982) Biochim. Biophys. Acta 720, 181187
27. Harder, T., Scheiffele, P., Verkade, P., and Simons, K. (1998) J. Cell Biol. 141,
929 942
28. Fridriksson, E. K., Shipkova, P. A., Sheets, E. D., Holowka, D., Baird, B., and
McLafferty, F. W. (1999) Biochemistry 38, 8056 8063
29. Field, K. A., Holowka, D., and Baird, B. (1997) J. Biol. Chem. 272, 4276 4280
30. Viola, A., Schroeder, S., Sakakibara, Y., and Lanzavecchia, A. (1999) Science
283, 680 682
31. Janes, P. W., Ley, S. C., and Magee, A. I. (1999) J. Cell Biol. 147, 447 461
32. Schutz, G. J., Kada, G., Pastushenko, V. P., and Schindler, H. (2000) EMBO J.
19, 892901
33. Mabrey, S., and Sturtevant, J. M. (1976) Proc. Natl. Acad. Sci. U. S. A. 73,
38623866
34. Korlach, J., Schwille, P., Webb, W. W., and Feigenson, G. W. (1999) Proc. Natl.
Acad. Sci. U. S. A. 96, 8461 8466
35. Bagatolli, L. A., and Gratton, E. (2000) Biophys. J. 78, 290 305
36. Varma, R., and Mayor, S. (1998) Nature 394, 798 802
37. Kenworthy, A. K., Petranova, N., and Edidin, M. (2000) Mol. Biol. Cell 11,
16451655
38. Schneiter, R., Brugger, B., Sandhoff, R., Zellnig, G., Leber, A., Lampl, M.,
Athenstaedt, K., Hrastnik, C., Eder, S., Daum, G., Paltauf, F., Wieland,
F. T., and Kohlwein, S. D. (1999) J. Cell Biol. 146, 741754
39. Mateo, C. R., Acuna, A. U., and Brochon, J.-C. (1995) Biophys. J. 68, 978 987
40. Schmidt, C. F., Barenholz, Y., Huang, C., and Thompson, T. E. (1978) Nature
271, 775777
41. Ren, J., Lew, S., Wang, Z., and London, E. (1997) Biochemistry 36,
1021310220
42. Ren, J., Lew, S., Wang, J., and London, E. (1999) Biochemistry 38, 59055912
43. Martens, J. R., Navarro-Polanco, R., Coppock, E. A., Nishiyama, A., Parshley,
L., Grobaski, T. D., and Tamkun, M. M. (2000) J. Biol. Chem. 275,
74437446
44. Weimbs, T., Low, S. H., Chapin, S. J., and Mostov, K. E. (1997) Trends Cell
Biol. 7, 393399
45. Rodriguez-Boulan, E., and Gonzalez, A. (1999) Trends Cell Biol. 9, 291294
46. Benting, J. H., Rietveld, A. G., and Simons, K. (1999) J. Cell Biol. 146, 313320
47. Mukherjee, S., Soe, T. T., and Maxfield, F. R. (1999) J. Cell Biol. 144,
12711284
48. Fivaz, M., Abrami, L., and van der Goot, F. (1999) Trends Cell Biol. 9, 212213
49. Green, J. M., Ahelesnyak, A., Chung, J., Lindberg, F. P., Sarfati, M., Frazier,
W. A., and Brown, E. J. (1999) J. Cell Biol. 146, 673 682
50. Krauss, K., and Altevogt, P. (1999) J. Biol. Chem. 274, 3692136927
51. Manes, S., Mira, E., Gomez-Mouton, C., Lacalle, R. A., Keller, P., Labrador,
J. P., and Martnez-A, C. (1999) EMBO J. 18, 6211 6220
52. Ravetch, J. V., and Clynes, R. A. (1998) Annu. Rev. Immunol. 16, 421 432
53. Montixi, C., Langlet, C., Bernard, A.-M., Thimonier, J., Dubois, C., Wurbel,
M.-A., Chauvin, J.-P., Pierres, M., and He, H.-T. (1998) EMBO J. 17,
5334 5348
54. Cheng, P. C., Dykstra, M. L., Mitchell, R. N., and Pierce, S. K. (1999) J. Exp.
Med. 190, 1549 1560
55. Lang, M. L., Shen, L., and Wade, W. F. (1999) J. Immunol. 163, 53915398
56. Sheets, E. D., Holowka, D., and Baird, B. (1999) J. Cell Biol. 145, 877 887
57. Harder, T., and Simons, K. (1999) Eur. J. Immunol. 29, 556 562
58. Xavier, R., and Seed, B. (1999) Curr. Opin. Immunol. 11, 265269
59. Ilangumaran, S., He, H.-T., and Hoessli, D. C. (2000) Immunol. Today 21, 27
60. Xavier, R., Brennan, T., Li, Q., McCormack, C., and Seed, B. (1998) Immunity
8, 723732
61. Moran, M., and Miceli, M. C. (1998) Immunity 9, 787796
62. Zhang, W., Sloan-Lancaster, J., Kitchen, J., Trible, R. P., and Samelson, L. E.
(1998) Cell 92, 8392
63. Lin, J., Weiss, A., and Finco, T. S. (1999) J. Biol. Chem. 274, 2886128864
64. Green, J. M., Schreiber, A. D., and Brown, E. J. (1997) J. Cell Biol. 139,
1209 1218
65. Chuang, F. Y. S., Sassaroli, M., and Unkeless, J. C. (2000) J. Immunol. 164,
350 360
66. Lou, Z., Jevremovic, D., Billadeau, D. D., and Leibson, P. J. (2000) J. Exp. Med.
191, 347354
67. Pike, L. J., and Miller, J. M. (1998) J. Biol. Chem. 273, 22298 22304
68. Furuchi, T., and Anderson, R. G. W. (1998) J. Biol. Chem. 273, 21099 21104