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Originally published In Press as doi:10.1074/jbc.M413993200 on May 19, 2005

J. Biol. Chem., Vol. 280, Issue 28, 26321-26329, July 15, 2005
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Headgroup-specific Exposure of Phospholipids in ABCA1-expressing Cells*

Nele Alder-Baerens{ddagger}§, Peter Müller{ddagger}, Antje Pohl{ddagger}, Thomas Korte{ddagger}, Yannick Hamon||, Giovanna Chimini||**, Thomas Pomorski{ddagger}, and Andreas Herrmann{ddagger}**{ddagger}{ddagger}

From the {ddagger}Humboldt University of Berlin, Institute of Biology, Center of Biophysics and Bioinformatics, Invalidenstrasse 42, 10115 Berlin, Germany and the ||Centre d'Immunologie de Marseille Luminy, Case 906, Parc Scientifique de Luminy, 13288 Marseille Cedex 09, France

ABCA1 has been established to be required for the efflux of cholesterol and phospholipids to apolipoproteins such as apoA-I. At present, it is unclear whether ABCA1-mediated lipid exposure is specific with regard to lipid headgroups and whether it requires calcium activation and the presence of a lipid acceptor. In the present work, we found exofacial exposure of endogenous phosphatidylserine in the absence of apoA-I to be enhanced in ABCA1-GFP expressing MDCKII and HeLa cells compared with control cells. By using C6-N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) (NBD)-labeled phospholipid analogues, we observed elevated redistribution of phosphatidylserine and phosphatidylethanolamine but not of phosphatidylcholine analogues from the cytoplasmic to the exoplasmic leaflet of the plasma membrane of ABCA1-GFP expressing cells. Whereas glyburide affected neither the level of exofacial endogenous PS nor the outward movement of the amino phospholipid analogues, the latter was sensitive to intracellular Ca2+ in ABCA1-GFP expressing cells, further enhancing outward analogue redistribution with respect to control cells. Both receptor-mediated endocytosis and fluidphase endocytosis were reduced in MDCKII cells expressing ABCA1-GFP. Glyburide raised the level of receptor-mediated endocytosis in the ABCA1-GFP expressing cell to the level of control cells in the absence of glyburide. In control cells, however, fluid-phase endocytosis but not receptor-mediated endocytosis was significantly reduced upon glyburide treatment.


Received for publication, December 13, 2004 , and in revised form, May 17, 2005.

* The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

§ Supported by a fellowship of the Schering Foundation.

Supported by a grant from the Deutsche Forschungsgemeinschaft.

** Supported by European Union Grant MRTN-CT-2004-005330.

{ddagger}{ddagger} To whom correspondence should be addressed: Humboldt University of Berlin, Institute of Biology, Molecular Biophysics, Invalidenstrasse 42, 10115 Berlin, Germany. Tel.: 49-30-2093-8830; Fax: 49-302093-8585; E-mail: andreas.herrmann{at}rz.hu-berlin.de.


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