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标题：A sensitive and specific ELISA detects methionine sulfoxide-containing apolipoprotein A-I in HDL
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作者：Xiao Suo Wang ; Baohai Shao ; Michael N. Oda 等
期刊名称：JLR Papers In Press
印刷版ISSN：0022-2275
电子版ISSN：1539-7262
出版年度：2009
卷号：50
期号：3
页码：586-594
DOI：10.1194/jlr.D800042-JLR200
语种：English
出版社：American Society for Biochemistry and Molecular Biology
摘要：Oxidized HDL has been proposed to play a key role in atherogenesis. A wide range of reactive intermediates oxidizes methionine residues to methionine sulfoxide (MetO) in apolipoprotein A-I (apoA-I), the major HDL protein. These reactive species include those produced by myeloperoxidase, an enzyme implicated in atherogenesis. The aim of the present study was to develop a sensitive and specific ELISA for detecting MetO residues in HDL. We therefore immunized mice with HPLC-purified human apoA-I containing MetO⁸⁶ and MetO¹¹² (termed apoA-I₊₃₂) to generate a monoclonal antibody termed MOA-I. An ELISA using MOA-I detected lipid-free apoA-I₊₃₂, apoA-I modified by 2 e -oxidants (hydrogen peroxide, hypochlorous acid, peroxynitrite), and HDL oxidized by 1 e - or 2 e -oxidants and present in buffer or human plasma. Detection was concentration dependent, reproducible, and exhibited a linear response over a physiologically plausible range of concentrations of oxidized HDL. In contrast, MOA-I failed to recognize native apoA-I, native apoA-II, apoA-I modified by hydroxyl radical or metal ions, or LDL and methionine-containing proteins other than apoA-I modified by 2 e -oxidants. Because the ELISA we have developed specifically detects apoA-I containing MetO in HDL and plasma, it should provide a useful tool for investigating the relationship between oxidized HDL and coronary artery disease.