Journal
JOURNAL OF LIPID RESEARCH
Volume 50, Issue 3, Pages 586-594Publisher
ELSEVIER
DOI: 10.1194/jlr.D800042-JLR200
Keywords
protein oxidation; lipid hydroperoxides; dysfunctional HDL; myeloperoxidase; oxidative stress; reactive nitrogen species
Categories
Funding
- Psiron Pty Ltd
- University of Sydney Postgraduate Award Scholarship
- National Health & Medical Research Council of Australia
- University of Sydney Medical Foundation
- National Institutes of Health [K99HL091055, HL086798, P30DK017047, PO1HL030086]
- Mass Spectrometry Core, Diabetes, and Endocrinology Research Center, University of Washington
Ask authors/readers for more resources
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 MetO(86) and MetO(112) (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 2e-oxidants (hydrogen peroxide, hypochlorous acid, peroxynitrite), and HDL oxidized by 1e- or 2e-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 2e-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.-Wang, X. S., B. Shao, M. N. Oda, J. W. Heinecke, S. Mahler, and R. Stocker. A sensitive and specific ELISA detects methionine sulfoxide-containing apolipoprotein A-I in HDL. J. Lipid Res. 2009. 50: 586-594.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available