Substrate and site specificity of hydrogen peroxide generation in mouse mitochondria
Substrate and site specificity of hydrogen peroxide generation in mouse mitochondria
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DOI：
10.1006/abbi.1997.0489
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发表时间：
1998-02-01
影响因子：
3.9
通讯作者：
Sohal, RS
中科院分区：
文献类型：
--
作者：
Kwong, LK;Sohal, RS
本研究的目的是阐明线粒体H2 O2产生的机制，在小鼠器官中，通过确定其性质的差异，底物利用，抑制剂的敏感性，和位点特异性影响H2 O2的生产。从心脏、脑和肾脏分离线粒体，并使用FADH连接的底物琥珀酸和α-甘油磷酸以及NADH连接的底物丙酮酸/苹果酸、β-羟基丁酸和谷氨酸测量H2 O2产生速率，呼吸抑制剂、抗霉素和鱼藤酮，单独并顺序添加到每个基质支持的H2 O2生成反应混合物中，以确定线粒体的生成位点和最佳条件（对于最大的Fates生成，琥珀酸支持最高的线粒体H_2O_2生成速率，而且，它是心脏线粒体的优选底物，α-甘油磷酸是心脏线粒体H_2O_2生成的不良底物。抑制剂研究表明，心脏线粒体对抗霉素最敏感，而脑对鱼藤酮最敏感。令人惊讶的发现是，肾脏线粒体中NADPH连接的底物支持的H_2O_2产生对鱼藤酮没有反应。线粒体H_2O_2产生的三个位点（泛醌、NADH脱氢酶和α-甘油磷酸脱氢酶）对总H_2O_2产生的贡献是底物和器官依赖的。结果表明，在比较不同器官之间线粒体H_2O_2产生的位点和速率之前，必须重新考虑测定条件。(C)北京：人民出版社.
The objective of this study was to elucidate the mechanisms of mitochondrial H2O2 generation in mouse organs by determining the nature of their differences in substrate utilization, inhibitor sensitivity, and the site specificity affecting H2O2 production. Mitochondria were isolated ft om heart, brain, and kidney and the rate of H2O2 generation was measured using the FADH-linked substrates succinate and alpha-glycerophosphate as well as the NADH-linked substrates pyruvate/malate, beta-hydroxybutyrate, and glutamate, Respiratory inhibitors, antimycin and rotenone, were added singly and sequentially to each substrate-supported H2O2 generation reaction mixture to determine the mitochondrial site(s) of generation and the optimal condition(s) for maximal Fates of generation, Succinate supported the highest rate of mitochondrial H2O2 generation, Moreover, it was the preferred substrate for the heart mitochondria, alpha-Glycerophosphate is a poor substrate for H2O2 generation ill heart mitochondria. Inhibitor studies showed that heart mitochondria were the most sensitive and responsive to antimycin, while brain was the most sensitive to rotenone, Fn surprising finding was that NADPH-linked substrate-supported H2O2 generation in kidney mitochondria mas not responsive to rotenone. The contribution from each of the three sites (ubiquinone, NADH dehydrogenase, and alpha-glycerophosphate dehydrogenase) of mitochondrial H2O2 generation to the total was both substrate and organ dependent, Results indicate that assay conditions must re considered before comparisons of sites and rates of mitochondrial H2O2 generation among different organs call be made. (C) 1988 Academic Press.