Nε-(carboxymethyl)lysine and 3-DG-imidazolone are major AGE structures in protein modification by 3-deoxyglucosone
Nε-(carboxymethyl)lysine and 3-DG-imidazolone are major AGE structures in protein modification by 3-deoxyglucosone
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DOI：
10.1093/jb/mvh124
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发表时间：
2004-09-01
影响因子：
2.7
通讯作者：
Horiuchi, S
中科院分区：
文献类型：
--
作者：
Jono, T;Nagai, R;Horiuchi, S
血浆3-脱氧葡萄糖（3-DG）水平在高血糖状态下升高，并与糖尿病并发症的发病机制有关，因为3-DG与蛋白质形成晚期糖基化终产物（AGE）的高反应性。为了研究体外和体内3- dg介导的蛋白质修饰的潜在标记物，我们通过免疫化学分析和高效液相色谱比较了几种3- dg衍生的AGE结构的产量，并测量了它们在人类动脉粥样硬化病变中的定位。3-DG与牛血清白蛋白37℃孵育4周后，N-epsilon-（羧甲基）赖氨酸（CML）和3-DG-咪唑酮的含量随孵育时间的增加而急剧增加，而吡啶和戊苷的含量在第28天略有增加。与3-DG在60°c下孵育时，也观察到大量的吡啶和戊苷，以促进age的形成。在动脉粥样硬化病变中，CML和3- dg -咪唑酮在大多数泡沫细胞的细胞质内和动脉粥样硬化核心的细胞外被发现。在动脉粥样硬化病变的主动脉内膜的细胞外基质和少量泡沫细胞中发现与吡咯碱的弱阳性免疫反应。我们的研究结果首次证明了CML和3-DG-咪唑酮是3-DG修饰蛋白的主要AGE结构，并且3-DG-咪唑酮比吡啶碱提供了更好的3-DG修饰蛋白的标记物。
The levels of plasma 3-deoxyglucosone (3-DG) increase under hyperglycemic conditions and are associated with the pathogenesis of diabetic complications because of the high reactivity of 3-DG with proteins to form advanced glycation end products (AGE). To investigate potential markers for 3-DG-mediated protein modification in vitro and in vivo, we compared the yield of several 3-DG-derived AGE structures by immunochemical analysis and HPLC and measured their localization in human atherosclerotic lesions. When BSA was incubated with 3-DG at 37 C for up to 4 wk, the amounts of N-epsilon-(carboxymethyl)lysine (CML) and 3-DG-imidazolone steeply increased with incubation time, whereas the levels of pyrraline and pentosidine increased slightly by day 28. In contrast, significants amount of pyrraline and pentosidine were also observed when BSA was incubated with 3-DG at 60degreesC to enhance AGE-formation. In atherosclerotic lesions, CML and 3-DG-imidazolone were found intracellularly in the cytoplasm of most foam cells and extracellularly in the atheromatous core. A weak-positive immunoreaction with pyrraline was found in the extracellular matrix and a few foam cells in aortic intima with atherosclerotic lesions. Our results provide the first evidence that CML and 3-DG-imidazolone are major AGE structures in 3-DG-modified proteins, and that 3-DG-imidazolone provides a better marker for protein modification by 3-DG than pyrraline.