Cisplatin Binding and Inactivation of Mitochondrial Glutamate Oxaloacetate Transaminase in Cisplatin-Induced Rat Nephrotoxicity
Cisplatin Binding and Inactivation of Mitochondrial Glutamate Oxaloacetate Transaminase in Cisplatin-Induced Rat Nephrotoxicity
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DOI：
10.1271/bbb.130172
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发表时间：
2013-08-01
影响因子：
1.6
通讯作者：
Yamashita, Tetsuro
中科院分区：
文献类型：
--
作者：
Ozaki, Taku;Ishiguro, Sei-ichi;Yamashita, Tetsuro
顺铂是一种广泛使用的化疗药物，但其使用受到与线粒体功能障碍相关的肾毒性的限制。由于其机制知之甚少，我们的目的是确定顺铂靶向的线粒体蛋白。我们从SD大鼠分离肾脏线粒体蛋白，并进行顺铂亲和柱层析。在SDS PAGE上检测洗脱的蛋白质，并进行凝胶内胰蛋白酶消化和LC-MS/MS分析。我们确定了谷草转氨酶（GOT）和线粒体苹果酸脱氢酶（MDH）。接下来，我们给SD大鼠腹腔注射顺铂诱导肾毒性，并测定酶的活性。结果表明，顺铂给药后第1天，线粒体GOT活性严重下降。三天后，我们还发现线粒体MDH活性下降。我们的研究结果表明，顺铂结合线粒体GOT，抑制其活性，导致线粒体功能障碍和随后的肾毒性。
Cisplatin is a widely used chemotherapeutic agent, but its use is limited by nephrotoxicity associated with mitochondrial dysfunction. Because its mechanisms are poorly understood, we aimed to identify the mitochondrial proteins targeted by cisplatin. We isolated renal mitochondrial proteins from Sprague-Dawley (SD) rats and performed cisplatin-affinity column chromatography. The proteins eluted were detected on SDS PAGE and subjected to in-gel tryptic digestion and LC-MS/MS analysis. We identified glutamate oxaloacetate transaminase (GOT) and mitochondrial malate dehydrogenase (MDH). Next, we administered cisplatin intraperitoneally to SD rats to induce nephrotoxicity and assayed the activities of the enzymes. The results indicated that cisplatin caused a severe decrease in mitochondrial GOT activity on day 1 after cisplatin administration. Three d later, we also identified a decrease in mitochondrial MDH activity. Our results indicate that cisplatin binds to mitochondrial GOT and inhibits its activity, causing mitochondrial dysfunction and subsequent nephrotoxicity.