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Originally published In Press as doi:10.1194/jlr.M300336-JLR200 on January 1, 2004

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Journal of Lipid Research, Vol. 45, 635-644, April 2004
Copyright © 2004 by American Society for Biochemistry and Molecular Biology

ABCA1 mediates concurrent cholesterol and phospholipid efflux to apolipoprotein A-I

Jonathan D. Smith1,*, Wilfried Le Goff*, Megan Settle*, Gregory Brubaker*, Christine Waelde{dagger}, Andrew Horwitz{dagger} and Michael N. Oda§

* Department of Cell Biology NC10, The Cleveland Clinic Foundation, Cleveland, OH 44195
{dagger} 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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