J. Lipid Res.
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A more recent version of this article appeared on October 1, 2005

Papers In Press, published online ahead of print July 16, 2005
J. Lipid Res., doi:10.1194/jlr.M500207-JLR200
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Submitted on May 20, 2005
Revised on July 5, 2005
Accepted on July 11, 2005

Redistribution of macrophage cholesterol ester hydrolase from cytoplasm to lipid droplets upon lipid loading

Bin Zhao, Bernard J. Fisher, Richard W. St.Clair, Lawrence L. Rudel, and Shobha Ghosh

Department of Internal Medicine, Virginia Commonwealth University, Richmond, VA 23298-0050

Corresponding Author: shobha{at}hsc.vcu.edu

Hydrolysis of intracellular cholesteryl esters (CE) represents the first step in removal of cholesterol from lipid-laden foam cells associated with atherosclerotic lesions. Neutral cholesterol ester hydrolase (CEH) catalyses this reaction and we have recently cloned the cDNA for the human macrophages CEH and demonstrated increased mobilization of intracellular CE droplets by CEH over-expression. The present study was undertaken to test the hypothesis that for CE hydrolysis, CEH must become associated with the surface of the cytoplasmic lipid droplets. Our data show redistribution of CEH from cytosol to lipid droplets upon lipid loading of human THP-1 macrophages. Depletion of triacylglycerol (TG) by incubation with acyl-CoA synthetase inhibitor Triacsin D had no effect on CEH association with the lipid droplets suggesting that CEH associates with mixed (CE+TG) as well as TG-depleted, CE droplets. However, CEH had 2.5 fold higher activity when mixed droplets were used as substrate in an in vitro assay consistent with the reported higher cholesterol efflux from cells containing mixed isotropic droplets. Perilipin as well as adipophilin, two lipid droplet-associated proteins, were also present on the lipid droplets in THP-1 macrophages. In conclusion, CEH associates with its intracellular substrate (lipid droplets) and hydrolyzes CE more efficiently from mixed droplets.


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