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Journal of Lipid Research, Vol. 48, 86-95, January 2007
Copyright © 2007 by American Society for Biochemistry and Molecular Biology
Centre for Clinical and Population Sciences, Nutrition and Metabolism Group, Queen's University, Belfast, United Kingdom
Published, JLR Papers in Press, October 25, 2006.
1 To whom correspondence should be addressed. e-mail: j.mceneny{at}qub.ac.uk
The plasma HDLs represent a major class of cholesterol-transporting lipoprotein that can be divided into two distinct subfractions, HDL2 and HDL3, by ultracentrifugation. Existing methods for the subfractionation of HDL requires lengthy ultracentrifugations, making them unappealing for large-scale studies. We describe a method that subfractionates HDL from plasma in only 6 h, representing a substantial decrease in total isolation time. The subfractions so isolated were assessed for a variety of lipid and protein components, in addition to their susceptibility to oxidation, both alone and in combination with VLDL and LDL. We report for the first time a prooxidant role for HDL during VLDL oxidation, in which HDL donates preformed hydroperoxides to VLDL in a cholesteryl ester transfer protein (CETP)-dependent process. Examination of the participation of HDL in LDL oxidation has reinforced its classic role as a potent antioxidant. Furthermore, we have also implicated the second major HDL-associated enzyme, LCAT, in these processes, whereby it acts as a potent prooxidant during VLDL oxidation but as an antioxidant during LDL oxidation. Thus, we have identified a potentially duplicitous role for HDL in the pathogenesis of atherosclerosis, attributable to both CETP and LCAT.
Supplementary key words apolipoproteins cholesteryl ester transfer protein lecithin:cholesterol acyltransferase lipid hydroperoxides single radial immunodiffusion transferrin ultracentrifugation
Abbreviations: apo[a], apolipoprotein [a]; apoA-I, apolipoprotein A-I; CETP, cholesteryl ester transfer protein; cHDL, crude high density lipoprotein; t1/2max, time at half maximum (an equivalent of lag time)
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