title: Chemical and immunochemical identification of propanoyllysine derived from oxidized n-3 polyunsaturated fatty acid.
abstract: It is known that n-3 polyunsaturated fatty acids (PUFAs), such as docosahexaenoic acid and eicosapentaenoic acid, are rapidly oxidized in vitro. Nvarepsilon-(propanoyl)lysine (propionyllysine, or PRL) is formed from the reaction of the oxidized products of n-3 PUFAs and lysine. To evaluate the oxidized n-3 PUFA-derived protein modifications in vivo, we have developed detection methods using a novel monoclonal antibody against PRL as well as liquid chromatography-mass spectrometry (LC/MS/MS). The antibody obtained specifically recognized PRL. A strong positive staining in atherosclerotic lesions of hypercholesterolemic rabbits was observed. We have also simultaneously identified and quantified both urinary PRL and urinary Nvarepsilon-(hexanoyl)lysine, using LC/MS/MS using isotope dilution methods. The level of urinary PRL (21.6+/-10.6 micromol/mol of creatinine) significantly correlated with the other oxidative stress markers, 8-oxo-deoxyguanosine, dityrosine, and isoprostanes. The increase in the excretion of amide adducts into the urine of diabetic patients was also confirmed compared to healthy subjects. These results suggest that PRL may be good marker for n-3 PUFA-derived oxidative stress in vivo.
authors.affiliation: Nagoya University Graduate School of Bioagricultural Sciences, Nagoya 464-8601, Japan.
authors.forename: Shinsuke
authors.initials: S
authors.lastname: Hisaka
authors.forename: Yoji
authors.initials: Y
authors.lastname: Kato
authors.forename: Noritoshi
authors.initials: N
authors.lastname: Kitamoto
authors.forename: Akihiro
authors.initials: A
authors.lastname: Yoshida
authors.forename: Yoshiko
authors.initials: Y
authors.lastname: Kubushiro
authors.forename: Michitaka
authors.initials: M
authors.lastname: Naito
authors.forename: Toshihiko
authors.initials: T
authors.lastname: Osawa
mesh_terms: D000080242:8-Hydroxy-2'-Deoxyguanosine; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001011:Aorta / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D015415:Biomarkers / Q000652:urine; D003324:Coronary Artery Disease / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D003849:Deoxyguanosine / Q000031:analogs & derivatives / Q000652:urine; D003919:Diabetes Insipidus / Q000503:physiopathology / Q000652:urine; D015237:Dinoprost / Q000031:analogs & derivatives / Q000652:urine; D005231:Fatty Acids, Unsaturated / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D006937:Hypercholesterolemia / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D007120:Immunochemistry; D008239:Lysine / Q000031:analogs & derivatives / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D013058:Mass Spectrometry; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D011422:Propionates / Q000737:chemistry* / Q000276:immunology* / Q000378:metabolism; D011817:Rabbits
chemical_list: D000911:Antibodies, Monoclonal; D015415:Biomarkers; D005231:Fatty Acids, Unsaturated; D011422:Propionates; C545748:propanoyllysine; C075750:8-epi-prostaglandin F2alpha; D000080242:8-Hydroxy-2'-Deoxyguanosine; D015237:Dinoprost; D003849:Deoxyguanosine; D008239:Lysine