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J. Biol. Chem., Vol. 276, Issue 27, 25324-25335, July 6, 2001
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From the Despite the critical role lipid droplets play in
maintaining energy reserves and lipid stores for the cell, little is
known about the regulation of the lipid or protein components within the lipid droplet. Although immunofluorescence of intact cells as well
as Western analysis of isolated lipid droplets revealed that sterol
carrier protein-2 (SCP-2) was not associated with lipid droplets, SCP-2
expression significantly altered the structure of the lipid droplet.
First, the targeting of fatty acid and cholesterol to the lipid
droplets was significantly decreased. Second, the content of several
proteins important for lipid droplet function was differentially
increased (perilipin A), reduced severalfold (adipose
differentiation-related protein (ADRP), vimentin), or almost completely
eliminated (hormone-sensitive lipase and proteins >93 kDa) in the
isolated lipid droplet. Third, the distribution of lipids within
the lipid droplets was significantly altered. Double labeling of cells
with
12-(N-methyl)-N-[(7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino]-octadecanoic
acid (NBD-stearic acid) and antisera to ADRP showed that 70, 24, and 13% of lipid droplets contained ADRP, NBD-stearic acid, or both,
respectively. SCP-2 expression decreased the level of ADRP in the lipid
droplet but increased the proportion wherein ADRP and NBD-stearic acid
colocalized by 3-fold. SCP-2 expression also decreased the lipid
droplet fatty acid and cholesterol mass (nmol/mg protein) by 5.2- and
6.6-fold, respectively. Finally, SCP-2 expression selectively altered
the pattern of esterified fatty acids in favor of polyunsaturated fatty
acids within the lipid droplet. Displacement studies showed
differential binding affinity of ADRP for cholesterol and fatty acids.
These data suggested that SCP-2 and ADRP play a significant role in
regulating fatty acid and cholesterol targeting to lipid droplets as
well as in determining their lipid and protein components.
Sterol Carrier Protein-2 Expression Modulates Protein
and Lipid Composition of Lipid Droplets*
,,,,,¶
Department of Physiology and Pharmacology
Texas A & M University, College Station, Texas 77843-4466 and the
§ Program in Obesity/Metabolism JM-USDA Human Nutrition
Research Center on Aging, Tufts University,
Boston, Massachusetts 02111
*
This work was supported in part by the United States Public
Health Service Grants DK41406 and GM31651 from the National Institutes of Health.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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