Oxidative stress promotes specific protein damage in Saccharomyces cerevisiae.
Oxidative stress promotes specific protein damage in Saccharomyces cerevisiae.
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DOI：
10.1074/jbc.m003140200
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发表时间：
2000-09
期刊：
影响因子：
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通讯作者：
E. Cabiscol;Eva Piulats;P. Echave;E. Herrero;J. Ros
中科院分区：
文献类型：
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作者：
E. Cabiscol;Eva Piulats;P. Echave;E. Herrero;J. Ros
我们已经分析了蛋白质的氧化损伤时，酿酒酵母细胞暴露于应力条件。通过蛋白质印迹法检测由过氧化氢或甲萘醌在有氧呼吸细胞的蛋白质上产生的羰基，纯化并鉴定。线粒体蛋白，如丙酮酸脱氢酶和α-酮戊二酸脱氢酶的E2亚基，乌头酸酶，热休克蛋白60，和细胞溶质脂肪酸合成酶（α亚基）和甘油醛-3-磷酸脱氢酶是主要的目标。此外，我们还报告了在测试的强制降解条件下，上述E2亚基中存在的硫辛酰胺的体内修饰，并且这也发生在用于比较分析的大肠杆菌细胞中存在的同源酶中。在发酵条件下，S.用过氧化氢或甲萘醌处理的酿酒酵母细胞的丙酮酸脱羧酶、烯醇酶、脂肪酸合成酶和甘油醛-3-磷酸脱氢酶。在测试的强制降解条件下，发酵细胞表现出比有氧呼吸细胞更低的活力，并且始终增加过氧化物生成以及更高的蛋白质羰基和脂质过氧化物含量。我们的研究结果有力地表明，在原核和真核细胞的氧化应激共享共同的特点。
We have analyzed the proteins that are oxidatively damaged when Saccharomyces cerevisiae cells are exposed to stressing conditions. Carbonyl groups generated by hydrogen peroxide or menadione on proteins of aerobically respiring cells were detected by Western blotting, purified, and identified. Mitochondrial proteins such as E2 subunits of both pyruvate dehydrogenase and alpha-ketoglutarate dehydrogenase, aconitase, heat-shock protein 60, and the cytosolic fatty acid synthase (alpha subunit) and glyceraldehyde-3-phosphate dehydrogenase were the major targets. In addition we also report the in vivo modification of lipoamide present in the above-mentioned E2 subunits under the stressing conditions tested and that this also occurs with the homologous enzymes present in Escherichia coli cells that were used for comparative analysis. Under fermentative conditions, the main protein targets in S. cerevisiae cells treated with hydrogen peroxide or menadione were pyruvate decarboxylase, enolase, fatty acid synthase, and glyceraldehyde-3-phosphate dehydrogenase. Under the stress conditions tested, fermenting cells exhibit a lower viability than aerobically respiring cells and, consistently, increased peroxide generation as well as higher content of protein carbonyls and lipid peroxides. Our results strongly suggest that the oxidative stress in prokaryotic and eukaryotic cells shares common features.