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Papers In Press, published online ahead of print August 1, 2006
J. Lipid Res., doi:10.1194/jlr.M500546-JLR200

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Journal of Lipid Research, Vol. 47, 1791-1802, August 2006
Copyright © 2006 by American Society for Biochemistry and Molecular Biology

Efflux of sphingomyelin, cholesterol, and phosphatidylcholine by ABCG1

Aya Kobayashi^*, Yasukazu Takanezawa, Takashi Hirata^*, Yuji Shimizu^*, Keiko Misasa^*, Noriyuki Kioka^*, Hiroyuki Arai, Kazumitsu Ueda^1,* and Michinori Matsuo^*

^* Laboratory of Cellular Biochemistry, Division of Applied Life Sciences, Kyoto University Graduate School of Agriculture, Kyoto 606-8502, Japan
Department of Health Chemistry, Graduate School of Pharmaceutical Sciences, University of Tokyo, Tokyo 113-0033, Japan

The online version of this article (available at http://www.jlr.org) contains an additional four figures.

Published, JLR Papers in Press, May 15, 2006.

¹ To whom correspondence should be addressed. e-mail: uedak{at}kais.kyoto-u.ac.jp

Cholesterol and phospholipids are essential to the body, but an excess of cholesterol or lipids is toxic and a risk factor for arteriosclerosis. ABCG1, one of the half-type ABC proteins, is thought to be involved in cholesterol homeostasis. To explore the role of ABCG1 in cholesterol homeostasis, we examined its subcellular localization and function. ABCG1 and ABCG1-K120M, a WalkerA lysine mutant, were localized to the plasma membrane in HEK293 cells stably expressing ABCG1 and formed a homodimer. A stable transformant expressing ABCG1 exhibited efflux of cholesterol and choline phospholipids in the presence of BSA, and the cholesterol efflux was enhanced by the presence of HDL, whereas cells expressing ABCG1-K120M did not, suggesting that ATP binding and/or hydrolysis is required for the efflux. Mass and TLC analyses revealed that ABCG1 and ABCA1 secrete several species of sphingomyelin (SM) and phosphatidylcholine (PC), and SMs were preferentially secreted by ABCG1, whereas PCs were preferentially secreted by ABCA1. These results suggest that ABCA1 and ABCG1 mediate the lipid efflux in different mechanisms, in which different species of phospholipids are secreted, and function coordinately in the removal of cholesterol and phospholipids from peripheral cells.

Abbreviations: apoA-I, apolipoprotein A-I; DSP, dithiobis (succinimidylpropionate); DTBP, dimethyl 3,3'-dithiobispropionimidate-HCl; LXR, liver X receptor; NBF, nucleotide binding fold; PC, phosphatidylcholine; RXR, retinoid X receptor; SM, sphingomyelin; sulfo-NHS-biotin, sulfo-N-hydroxysuccinimidobiotin

Supplementary key words ATP binding cassette protein G1 • ATP binding cassette protein A1 • high density lipoprotein

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