report

Rafts defined: a report on the Keystone symposium on

lipid rafts and cell function
Linda J. Pike1
Washington University School of Medicine, Department of Biochemistry and Molecular Biophysics,
St. Louis, MO 63110

Abstract The recent Keystone Symposium on Lipid Rafts invited to contribute to this effort, and the work product
and Cell Function (March 23–28, 2006 in Steamboat reflects the consensus of this broad-based group.
Springs, CO) brought together biophysicists, biochemists, The definition adopted by the group was as follows:
and cell biologists to discuss the structure and function of
“Membrane rafts are small (10–200 nm), heterogeneous,
lipid rafts. What emerged from the meeting was a consensus
definition of a membrane raft: “Membrane rafts are small highly dynamic, sterol- and sphingolipid-enriched do-
(10–200 nm), heterogeneous, highly dynamic, sterol- and mains that compartmentalize cellular processes. Small

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sphingolipid-enriched domains that compartmentalize cel- rafts can sometimes be stabilized to form larger platforms
lular processes. Small rafts can sometimes be stabilized to through protein-protein and protein-lipid interactions.”
form larger platforms through protein-protein and protein- This definition was arrived at by listing all possible terms
lipid interactions.”” This definition helps to clarify current that could be used to describe lipid rafts, discussing and
thinking in a field that has been plagued by the heteroge- prioritizing them, and then working them into a definition
neous and sometimes ephemeral nature of its subject.— for these domains. The terms that did not make it into the
Pike, L. J. Rafts defined: a report on the Keystone sympo-
sium on lipid rafts and cell function. J. Lipid Res. 2006. 47: definition are at least as revealing of the state of the field as
1597–1598. are the terms that did make the final cut. The definition is
intended to apply specifically to microdomains in cells, not
Supplementary key words membrane microdomains . cholesterol .
in model membranes, which are thought to be governed
sphingolipids by a different, but overlapping, set of rules.
First and foremost, the term “lipid raft” was discarded in
favor of the term “membrane raft.” Although phase separa-
It is rare that a meeting is held to discuss an entity that is tion of lipids is acknowledged to provide an underlying
difficult to visualize, has an ill-defined molecular compo- energetic drive for the formation of membrane domains,
sition, and whose very existence has been questioned (1). the concept that the formation of membrane rafts is deter-
Such was the case in Steamboat Springs, CO, on March 23– mined solely by lipid-driven interactions has been sup-
28, 2006, when the Keystone Symposium on Lipid Rafts planted by the understanding that proteins and lipids both
and Cell Function was convened. Organized by Linda Pike contribute to the genesis of these membrane microdomains.
(Washington University) and Michael Edidin ( Johns Although “plasma membrane” was suggested for in-
Hopkins University), the meeting brought together bio- clusion in the definition of membrane rafts, it was quickly
physicists, biochemists, and cell biologists to ponder the excluded from consideration. Presentations and posters
question, “What is a raft?” indicating the existence of raft-like domains on intracel-
Presentations ranged from the very biophysical talk lular membranes such as the endoplasmic reticulum and
given by Sarah Veatch (University of British Columbia) on mitochondria made it clear that the plasma membrane
phase behavior in model membranes to the biology-driven does not hold a monopoly on membrane domains. Al-
keynote address on the role of membrane domains in though the nature of such “non-plasma membrane” mem-
animal virus entry given by Ari Helenius (Institute of Bio- brane domains is not yet known, room must be left in the
chemistry, Eidgenössisch Technische Hochschule, Zurich, tent to welcome these newcomers.
Switzerland). Together, the discussions permitted the gen- This brings up the question of the typical lipid com-
eration of a definition for “lipid rafts” in an ad hoc session position of a membrane raft. Although enrichment in
on the final day of the meeting. All participants were cholesterol and sphingolipids was readily adopted as a
characteristic of membrane rafts, this composition could

Manuscript received 18 April 2006 and in revised form 27 April 2006.

1
Published, JLR Papers in Press, April 27, 2006. To whom correspondence should be addressed.
DOI 10.1194/jlr.E600002-JLR200 e-mail: [email protected]

Copyright D 2006 by the American Society for Biochemistry and Molecular Biology, Inc.

This article is available online at http://www.jlr.org Journal of Lipid Research Volume 47, 2006 1597
be difficult to achieve in intracellular membranes that copy, this methodology by itself cannot be used to co-
contain little if any of these two lipids. However, in her localize proteins or lipids to the same membrane raft.
talk, Sarah Veatch noted that rather than disrupting lipid The notion of raft heterogeneity has long been accepted
domains, cholesterol extraction from cells can increase in the field (6) for a host of reasons, including the fact that
the size of domains. Thus, it may be possible to generate proteins and lipids that are isolated together in the “raft”
membrane rafts in the absence of high concentrations fraction of cells are actually localized in different areas of
of cholesterol. the cell (7). It is now understood that this heterogeneity is
In the context of the lipid composition of membrane not limited to three-dimensional space but extends into
rafts, the term “liquid-ordered” was considered and dis- the fourth dimension, time. Rafts may be transient or re-
missed, largely on the grounds that there is, as yet, no solid latively stable, but all are viewed as dynamic structures.
evidence that cellular membrane rafts exist in this state. Several speakers ( John Hancock, University of Queens-
In a related judgment, the adjective “detergent-resistant” land; Anne Kenworthy, Vanderbilt University; Ken Jacob-
was considered but was soundly defeated. This outcome son, University of North Carolina; Satyajit Mayor, National
reflects the assessment of those in the field that detergent Centre for Biological Sciences, Bangalore, India) pre-
resistance is an artificial and highly subjective approach sented evidence that the concept of stable, preexisting
that can induce the formation of membrane domains and lipid rafts is no longer tenable. Clustering of some proteins
hence does not provide physiologically relevant informa- may occur because of differential affinity for or mobility in
tion (2). Some individual proteins or complexes that can membrane microdomains. Alternatively, rafts may arise
be shown by other methods to exist in rafts can be isolated from the stabilization of otherwise transient, nanoscale
through detergent-extraction procedures, but this ap- domains, often in an actin-dependent manner. The con-
proach for the de novo identification of raft components cept of the stabilization of rafts to form larger platforms
is no longer viable. actually arose from discussions of raft size and the ac-

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It came as no surprise that the term “small” was quickly knowledgment that some large domains, such as the immu-
brought up, and accepted, as a descriptor of rafts. Rafts have nological synapse or the entire brush border membrane,
been getting smaller as the methods applied to study them represent special cases in which small rafts are brought
have become more varied and more sophisticated. A host of together, through protein-protein interactions, to pro-
imaging and analytical methods used to study membrane duce a stable, biologically important structure.
rafts were reported at the meeting. Included among them A general consensus was that function was an important
were electron microscopy alone or coupled with spatial aspect of “raftness.” Many different functional roles for
statistics, heterofluorescence and homofluorescence reso- rafts were described by speakers and in posters at the
nance energy transfer, fluorescence quenching, fluores- meeting (viral and toxin entry, cellular signaling, and
cence lifetime imaging microscopy, fluorescence recovery protein and lipid trafficking). Although such specific
after photobleaching, fluorescence correlation spectrosco- functions were considered for inclusion in the definition
py, raster scan image correlation spectroscopy, and single of rafts, they were ultimately deemed to be too limiting.
particle tracking. The application of physical techniques The idea that rafts are involved in the compartmentaliza-
that are capable of analyzing complexes in the range of tens tion of cellular processes was felt to be more generally
of nanometers or less is an indication that the field has applicable and underscored the sense that, ultimately, it is
moved to a “less-is-more” view of membrane rafts. functional significance that underlies the importance of
But not too much less. Discussion swirled around the being a raft.
issue of how small (or how large) a raft can be. Complexes
in the range of only a few nanometers, referred to as “lipid
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