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Vol. 11, Issue 5, 1645-1655, May 2000
and
*Department of Biology, The Johns Hopkins University,
Baltimore, Maryland 21218; and "Lipid rafts" enriched in glycosphingolipids (GSL),
GPI-anchored proteins, and cholesterol have been proposed as functional microdomains in cell membranes. However, evidence supporting their existence has been indirect and controversial. In the past year, two
studies used fluorescence resonance energy transfer (FRET) microscopy
to probe for the presence of lipid rafts; rafts here would be defined
as membrane domains containing clustered GPI-anchored proteins at the
cell surface. The results of these studies, each based on a single
protein, gave conflicting views of rafts. To address the source of this
discrepancy, we have now used FRET to study three different
GPI-anchored proteins and a GSL endogenous to several different cell
types. FRET was detected between molecules of the GSL GM1
labeled with cholera toxin B-subunit and between antibody-labeled
GPI-anchored proteins, showing these raft markers are in submicrometer
proximity in the plasma membrane. However, in most cases FRET
correlated with the surface density of the lipid raft marker, a result
inconsistent with significant clustering in microdomains. We conclude
that in the plasma membrane, lipid rafts either exist only as
transiently stabilized structures or, if stable, comprise at most a
minor fraction of the cell surface.
Laboratory of
Leucocytes Antigens, Institute of Molecular Genetics, Academy of
Sciences of the Czech Republic, Videnská 1083, 14220 Prague,
Czech Republic
Corresponding author and present address:
Cell Biology and Metabolism Branch, National Institute of Child Health
and Human Development, National Institutes of Health, Building 18 T, Room 101, Bethesda, MD 20892. E-mail address: kenworta{at}mail.nih.gov.
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|
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|
|
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|
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|
|
|
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|
|
|
|
|
|
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|
|
|
|
|
|
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|
|
|
|
|
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|
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|
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|
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|
|
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|
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|
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|
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