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J Biol Chem, Vol. 273, Issue 41, 26338-26348, October 9, 1998
From the Department of Biology, Massachusetts Institute of
Technology, Cambridge, Massachusetts 02139
The class B type I scavenger
receptor, (SR-BI), is a member of the CD36 superfamily of proteins and
is a physiologically relevant, high affinity cell surface high density
lipoprotein (HDL) receptor that mediates selective lipid uptake. The
mechanism of selective lipid uptake is fundamentally different from
that of classic receptor-mediated uptake via coated pits and vesicles
(e.g. the low density lipoprotein receptor pathway) in that
it involves efficient transfer of the lipids, but not the outer shell
proteins, from HDL to cells. The abilities of SR-BI and CD36, both of
which are class B scavenger receptors, to bind HDL and mediate cellular
uptake of HDL-associated lipid when transiently expressed in COS cells
were examined. For these experiments, the binding of HDL to cells was
assessed using either 125I- or Alexa (a fluorescent
dye)-HDL in which the apolipoproteins on the surface of the HDL
particles were covalently modified. Lipid transfer was measured using
HDL noncovalently labeled by the fluorescent lipid
1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate.
Although both mSR-BI and human CD36 (hCD36) could mediate the binding
of HDL in a punctate pattern across the surfaces of cells, only mSR-BI
efficiently mediated the transfer of lipid to the cells. Analysis of
point mutants established that the major sites of fatty acylation of
mSR-BI are Cys462 and Cys470 and that fatty
acylation is not required for receptor clustering, HDL binding, or
efficient lipid transfer. Generation of mSR-BI/hCD36 domain swap
chimeras showed that the differences in lipid uptake activities between
mSR-BI and hCD36 were not due to differences between their two sets of
transmembrane and cytoplasmic domains but rather result from
differences in their large extracellular loop domains. These results
show that high affinity binding to a cell surface receptor is not
sufficient to ensure efficient cellular lipid uptake from HDL. Thus,
SR-BI-mediated binding combined with SR-BI-dependent
facilitated transfer of lipid from the HDL particle to the cell appears
to be the most likely mechanism for the bulk of the selective uptake of
cholesteryl esters from HDL to the liver and steroidogenic tissues.
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