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Journal of Lipid Research, Vol. 47, 1025-1036, May 2006
Copyright © 2006 by American Society for Biochemistry and Molecular Biology

Regulation of reconstituted high density lipoprotein structure and remodeling by apolipoprotein E

Kerry-Anne Rye^1,*,,, Richard Bright^*, Maria Psaltis^* and Philip J. Barter^*,

^* Lipid Research Group, The Heart Research Institute, Camperdown, Sydney, New South Wales, Australia 2050
Department of Medicine, University of Sydney, New South Wales, Australia 2006
Department of Medicine, University of Melbourne, Victoria, Australia 3010

Published, JLR Papers in Press, February 1, 2006.

¹ To whom correspondence should be addressed. e-mail: ryek{at}hri.org.au

Apolipoprotein E (apoE) enters the plasma as a component of discoidal HDL and is subsequently incorporated into spherical HDL, most of which contain apoE as the sole apolipoprotein. This study investigates the regulation, origins, and structure of spherical, apoE-containing HDLs and their remodeling by cholesteryl ester transfer protein (CETP). When the ability of discoidal reconstituted high density lipoprotein (rHDL) containing apoE2 [(E2)rHDL], apoE3 [(E3)rHDL], or apoE4 [(E4)rHDL] as the sole apolipoprotein to act as substrates for LCAT were compared with that of discoidal rHDL containing apoA-I [(A-I)rHDL], the rate of cholesterol esterification was (A-I)rHDL >> (E2)rHDL (E3)rHDL > (E4)rHDL. LCAT also had a higher affinity for discoidal (A-I)rHDL than for the apoE-containing rHDL. When the discoidal rHDLs were incubated with LCAT and LDL, the resulting spherical (E2)rHDL, (E3)rHDL, and (E4)rHDL were larger than, and structurally distinct from, spherical (A-I)rHDL. Incubation of the apoE-containing spherical rHDL with CETP and Intralipid® generated large fusion products without the dissociation of apoE, whereas the spherical (A-I)rHDLs were remodeled into small particles with the formation of lipid-poor apoA-I. In conclusion, i) apoE activates LCAT less efficiently than apoA-I; ii) apoE-containing spherical rHDLs are structurally distinct from spherical (A-I)rHDL; and iii) the CETP-mediated remodeling of apoE-containing spherical rHDL differs from that of spherical (A-I)rHDL.

Supplementary key words cholesteryl ester transfer protein • apolipoprotein A-I • kinetics

Abbreviations: apoE, apolipoprotein E; CE, cholesteryl ester; CETP, cholesteryl ester transfer protein; DPH, 1,6-diphenyl-1,3,5-hexatriene; Gdn-HCl, guanidine hydrochloride; PRODAN, 6-propionyl-2-(dimethylamino)-naphthalene; rHDL, reconstituted high density lipoprotein; Sulfo-EGS, sulfo-ethylene glycol bis[sulfosuccinimidylsuccinate]; TG, triglyceride; TMA-DPH, 1-(4-trimethylammoniumphenyl)-6-phenyl-1,3,5-hexatriene p-toluenesulfonate; UC, unesterified cholesterol

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