Adductome-based identification of biomarkers for lipid peroxidation.
Lipid peroxidation is an endogenous source of aldehydes that gives rise to covalent modification of proteins in various pathophysiological states. In this study, a strategy for the comprehensive detection and comparison of adducts was applied to find a biomarker for lipid peroxidation-modified proteinsin vivoThis adductome approach utilized liquid chromatography with electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) methods designed to detect the specific product ions from positively ionized adducts in a selected reaction monitoring mode. Using this procedure, we comprehensively analyzed lysine and histidine adducts generated in thein vitrooxidized low-density lipoproteins (LDL) and observed a prominent increase in several adducts, including a major lysine adduct. Based on the high resolution ESI-MS of the adduct and on the LC-ESI-MS/MS analysis of the synthetic adduct candidates, the major lysine adduct detected in the oxidized LDL was identified asNϵ-(8-carboxyoctanyl)lysine (COL). Strikingly, a significantly higher amount of COL was detected in the sera from atherosclerosis-prone mice and from patients with hyperlipidemia compared with the controls. These data not only offer structural insights into protein modification by lipid peroxidation products but also provide a platform for the discovery of biomarkers for human diseases.
Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya 464-8601; PRESTO, Japan Science and Technology Agency, Kawaguchi, Saitama 332-0012.
Takahiro
T
Shibata
Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya 464-8601.
Kazuma
K
Shimizu
Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya 464-8601.
Keita
K
Hirano
Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya 464-8601.
Fumie
F
Nakashima
Departments of Medical Technique.
Ryosuke
R
Kikuchi
Clinical Laboratory and Blood Transfusion, Nagoya University Hospital, Nagoya 466-8560.
Tadashi
T
Matsushita
Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya 464-8601; Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo 113-8657, Japan. Electronic address: a-uchida@mail.ecc.u-tokyo.ac.jp.
Koji
K
Uchida
D000818:Animals; D015415:Biomarkers / Q000097:blood; D005260:Female; D006801:Humans; D015227:Lipid Peroxidation; D008077:Lipoproteins, LDL / Q000097:blood*; D008239:Lysine / Q000031:analogs & derivatives / Q000097:blood; D008297:Male; D013058:Mass Spectrometry; D051379:Mice
D015415:Biomarkers; D008077:Lipoproteins, LDL; C099252:oxidized low density lipoprotein; D008239:Lysine
aldehyde; biomarker; lipid oxidation; lipoprotein; mass spectrometry (MS); protein chemical modification