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Journal of Lipid Research, Vol. 48, 882-889, April 2007
Copyright © 2007 by American Society for Biochemistry and Molecular Biology

Spontaneous reconstitution of discoidal HDL from sphingomyelin-containing model membranes by apolipoprotein A-I

Masakazu Fukuda^*, Minoru Nakano^1,*, Supaporn Sriwongsitanont, Masaharu Ueno, Yoshihiro Kuroda^* and Tetsurou Handa^*

^* Graduate School of Pharmaceutical Sciences, Kyoto University, Sakyo-ku, Kyoto 606-8501, Japan
Faculty of Pharmaceutical Sciences, Toyama University, 2630 Sugitani, Toyama 930-0194, Japan

Published, JLR Papers in Press, January 15, 2007.

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

Nascent HDL is known to be formed by the interaction of apolipoprotein A-I (apoA-I) with transmembrane ABCA1, but the molecular mechanism by which nascent HDL forms is less well understood. Here, we studied how reconstituted high density lipoprotein (rHDL) forms spontaneously on the interaction of apoA-I with model membranes. The formation of rHDL from pure phosphatidylcholine (PC) large unilamellar vesicles (LUVs) proceeded very slowly at 37.0°C, but sphingomyelin (SM) -rich PC/SM LUVs, which are in a gel/liquid-disordered phase (L_d phase) at this temperature, were rapidly microsolubilized to form rHDL by apoA-I. The addition of cholesterol decreased the rate at which rHDL formed and induced the selective extraction of lipids by apoA-I, which preferably extracted lipids of L_d phase rather than lipids of liquid-ordered phase. In addition, apoA-I extracted lipids from the outer and inner leaflets of LUVs simultaneously. These results suggest that the heterogeneous interface of the mixed membranes facilitates the insertion of apoA-I and induces L_d phase-selective but leaflet-nonselective lipid extraction to form rHDL; they are compatible with recent cell works on apoA-I-dependent HDL generation.

Supplementary key words ATP binding cassette transporter A1 • cholesterol • lipid efflux • liposome • liquid-disordered phase • liquid-ordered phase • phosphatidylcholine • high density lipoprotein

Abbreviations: apoA-I, apolipoprotein A-I; Chol, cholesterol; DPH, 1,6-diphenyl-1,3,5-hexatriene; L_d phase, liquid-disordered phase; L_o phase, liquid-ordered phase; LUV, large unilamellar vesicle; NBD-DPPE, 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(7-nitro-2-1,3-benzoxadiazol-4-yl); NBD-PC, 2-(6-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-hexanoyl-1-hexadecanoyl-sn-glycero-3-phosphocholine; PC, phosphatidylcholine; PL, phospholipid; pyrene-PC, 1-hexadecanoyl-2-(1-pyrenehexanoyl)-sn-glycero-3-phosphocholine; rHDL, reconstituted high density lipoprotein; SM, sphingomyelin

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