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* The Heart Research Institute, 145 Missenden Road, Camperdown NSW 2050, Australia
Prince of Wales Hospital, Randwick NSW 2031, Australia
Department of Pathology, School of Medical Sciences, Faculty of Medicine, The University of New South Wales, UNSW Sydney NSW 2052, Australia
** Centre for Thrombosis and Vascular Research, School of Medical Sciences, Faculty of Medicine, The University of New South Wales, UNSW Sydney NSW 2052, Australia
1 To whom correspondence should be addressed. e-mail: r.stocker{at}unsw.edu.au
Atherosclerosis is a state of heightened oxidative stress. Oxidized LDL is present in atherosclerotic lesions and used as marker for coronary artery disease, although in human lesions lipids associated with HDL are as oxidized as those of LDL. Here we investigated specific changes occurring to apolipoprotein A-I (apoA-I) and apoA-II, as isolated HDL and human plasma undergo mild, chemically induced oxidation, or autoxidation. During such oxidation, Met residues in apoA-I and apoA-II become selectively and consecutively oxidized to their respective Met sulfoxide (MetO) forms that can be separated by HPLC. Placing plasma at -20°C prevents autoxidation, whereas metal chelators and butylated hydroxytoluene offer partial protection. Independent of the oxidation conditions, apoA-I and apoA-II (dimer) with two MetO residues accumulate as relatively stable oxidation products. Compared to controls, serum samples from subjects with the endothelial cell nitric oxide synthase a/b genotype that is associated with increased coronary artery disease contain increased concentrations of apoA-I with two MetO residues.
Our results show that during the early stages, oxidation of HDL gives rise to specifically oxidized forms of apoA-I and apoA-II, some of which may be useful markers of in vivo HDL oxidation, and hence potentially atherosclerosis.
Abbreviations: AAPH, 2,2'-azo-bis(2-amidinopropane) dihydrochloride; BHT, butylated hydroxy toluene; DTPA, diethylenetriamine pentaacetic acid; eNOS, endothelial nitric oxide synthase; MetO, Met sulfoxide; oxLDL, oxidized low-density lipoprotein
Supplementary key words diagnostic • methonine sulfoxide • oxidative stress
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