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Journal of Lipid Research, Vol. 45, 635-644, April 2004
Copyright © 2004 by American Society for Biochemistry and Molecular Biology
* Department of Cell Biology NC10, The Cleveland Clinic Foundation, Cleveland, OH 44195
Laboratory of Biochemical Genetics and Metabolism, Rockefeller University, New York, NY 10021
Children's Hospital Oakland Research Institute, Oakland, CA 94609
1 To whom correspondence should be addressed. e-mail: smithj{at}lerner.ccf.org
in re-
Prior studies provide data supporting the notion that ATP binding cassette transporter A1 (ABCA1) promotes lipid efflux to extracellular acceptors in a two-step process: first, ABCA1 mediates phospholipid efflux to an apolipoprotein, and second, this apolipoprotein-phospholipid complex accepts free cholesterol in an ABCA1-independent manner. In the current study using RAW264.7 cells, ABCA1-mediated free cholesterol and phospholipid efflux to apolipoprotein A-I (apoA-I) were tightly coupled to each other both temporally and after treatment with ABCA1 inhibitors. The time course and temperature dependence of ABCA1-mediated lipid efflux to apoA-I support a role for endocytosis in this process. Cyclodextrin treatment of RAW264.7 cells partially inhibited 8Br-cAMP-induced efflux of free cholesterol and phospholipid to apoA-I.
ABCA1-expressing cells are more sensitive to cell damage by high-dose cyclodextrin and vanadate, leading to increased lactate dehydrogenase leakage and phospholipid release even in the absence of the acceptor apoA-I. Finally, we could not reproduce a two-step effect on lipid efflux using conditioned medium from ABCA1-expressing cells pretreated with cyclodextrin.
Abbreviations: AcLDL, acetylated LDL; DGGB, DMEM supplemented with 20 mM glucose, 2 mM glutamine, and 0.2% BSA; FC, free cholesterol; PL, phospholipid
Supplementary key words lipid efflux • ATP binding cassette transporter A1 • Tangier disease • reverse cholesterol transport • endocytosis • cyclodextrin
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