Immuno-spin trapping of a post-translational carboxypeptidase B1 radical formed by a dual role of xanthine oxidase and endothelial nitric oxide synthase in acute septic mice
Immuno-spin trapping of a post-translational carboxypeptidase B1 radical formed by a dual role of xanthine oxidase and endothelial nitric oxide synthase in acute septic mice
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DOI：
10.1016/j.freeradbiomed.2008.10.046
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发表时间：
2009-02-15
影响因子：
7.4
通讯作者：
Mason, Ronald P.
中科院分区：
文献类型：
--
作者：
Chatterjee, Saurabh;Ehrenshaft, Marilyn;Mason, Ronald P.
由于暴露于自由基和其他活性物质而导致的蛋白质翻译后修饰是代谢和炎症氧化应激（例如败血症）的标志。本研究使用硝酮自旋捕获 DMPO 并结合免疫自旋捕获和质谱分析来鉴定小鼠体内自由基反应的产物。我们报告了在用脂多糖 (LPS) 处理的小鼠脾脏中检测到 DMPO-羧肽酶 B1 (CPB1) 加合物的剂量依赖性产生。此外，我们报告在正常生理条件下的小鼠中显着检测到 DMPO-CPBI 加合物。抑制剂处理和基因敲除小鼠实验表明，黄嘌呤氧化酶和内皮一氧化氮合酶是导致 CPBI 加合物形成的活性物质的重要来源。我们还报告了 LPS 攻击后 CPB1 活性显着丧失，同时 CPBI 蛋白积累增加。这表明存在 CPBI 活性丧失和补偿性蛋白质产生的可能机制。由爱思唯尔公司出版
Post-translational modification of proteins due to exposure to radicals and other reactive species are markers of metabolic and inflammatory oxidative stress Such as sepsis. This study uses the nitrone spin-trap DMPO and a combination of immuno-spin trapping and mass spectrometry to identify in vivo products of radical reactions in mice. We report the detection of dose-dependent production of DMPO-carboxypeptidase B1 (CPB1) adducts in the spleens of mice treated with lipopolysaccharide (LPS). Additionally, we report significant detection of DMPO-CPBI adducts in mice experiencing normal physiological conditions. Treatments with inhibitors and experiments with knock-out Mice indicate that xanthine oxidase and endothelial nitric oxide synthase are important sources of the reactive species that lead to CPBI adduct formation. We also report a significant loss of CPB1 activity following LPS challenge in conjunction with an increase in CPBI protein accumulation. This Suggests the presence of a possible mechanism for CPBI activity loss with compensatory protein production. Published by Elsevier Inc.