Chemical and immunochemical identification of propanoyllysine derived from oxidized n-3 polyunsaturated fatty acid
Chemical and immunochemical identification of propanoyllysine derived from oxidized n-3 polyunsaturated fatty acid
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DOI：
10.1016/j.freeradbiomed.2009.02.030
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发表时间：
2009-06-01
影响因子：
7.4
通讯作者：
Osawa, Toshihiko
中科院分区：
文献类型：
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作者：
Hisaka, Shinsuke;Kato, Yoji;Osawa, Toshihiko
已知n-3多不饱和脂肪酸（PUFA），例如二十二碳六烯酸和二十碳五烯酸，在体外被快速氧化。N-（丙酰基）赖氨酸（propionyllysine，或PRL）由n-3 PUFA和赖氨酸的氧化产物的反应形成。为了评估，氧化的n-3 PUFA衍生的蛋白质修饰在体内，我们已经开发了检测方法，使用一种新的单克隆抗体对PRL以及液相色谱-质谱（LC/MS/MS）。所获得的抗体特异性识别PRL。在高胆固醇血症家兔的动脉粥样硬化病变中观察到强阳性染色。我们还同时确定和定量尿PRL和尿N-（己酰）赖氨酸，使用LC/MS/MS同位素稀释法。尿PRL水平（21.6 +/- 10.6 μ mol/mol肌酐）与其他氧化应激标志物8-氧代脱氧鸟苷、二酪氨酸显著相关。和异前列烷。与健康受试者相比，也证实了糖尿病患者尿液中酰胺加合物排泄量的增加。这些结果表明，PRL可能是体内n-3 PUFA衍生的氧化应激的良好标志物。(C)2009 Elsevier Inc. All rights reserved.
It is known that n-3 polyunsaturated fatty acids (PUFAs), such as docosahexaenoic acid and eicosapentaenoic acid, are rapidly oxidized ill vitro. N epsilon-(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 N epsilon-(hexanoyl) lysine, using LC/MS/MS using isotope dilution methods. The level of urinary PRL (21.6 +/- 10.6 mu mol/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. (C) 2009 Elsevier Inc. All rights reserved.