Using Quantitative Redox Proteomics to Dissect the Yeast Redoxome
Using Quantitative Redox Proteomics to Dissect the Yeast Redoxome
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DOI：
10.1074/jbc.m111.296236
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发表时间：
2011-12-02
影响因子：
4.8
通讯作者：
Jakob, Ursula
中科院分区：
文献类型：
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作者：
Brandes, Nicolas;Reichmann, Dana;Jakob, Ursula
为了理解并最终预测改变氧化还原条件和氧化剂水平对生物体生理学的影响，必须了解其氧化还原体：其活性受其半胱氨酸巯基氧化状态控制的蛋白质。在这里，我们应用定量氧化还原蛋白质组学方法OxICAT的酿酒酵母，并确定在体内的巯基氧化状态的近300种不同的酵母蛋白分布在不同的细胞室。我们发现，大量的细胞溶质和线粒体蛋白在指数增长过程中部分氧化。我们的研究结果表明，占主导地位的氧化还原条件不断控制中央细胞途径微调氧化状态，因此这些蛋白质的活性。亚致死浓度的H2O2处理导致41种蛋白质的亚组发生大量巯基修饰，从而影响各种不同的细胞途径，其中许多直接或间接参与增加氧化应激抗性。根据其稳态氧化水平和对过氧化物处理的敏感性对所识别的蛋白质硫醇进行分类，发现蛋白质硫醇的氧化还原敏感性不能预测过氧化物敏感性。我们的研究提供了实验证据，表明蛋白质硫醇对过氧化物水平变化的反应能力可能受热力学和动力学参数的影响，这使得预测硫醇修饰具有挑战性，并且对过氧化物敏感的蛋白质硫醇的从头鉴定不可或缺。
To understand and eventually predict the effects of changing redox conditions and oxidant levels on the physiology of an organism, it is essential to gain knowledge about its redoxome: the proteins whose activities are controlled by the oxidation status of their cysteine thiols. Here, we applied the quantitative redox proteomic method OxICAT to Saccharomyces cerevisiae and determined the in vivo thiol oxidation status of almost 300 different yeast proteins distributed among various cellular compartments. We found that a substantial number of cytosolic and mitochondrial proteins are partially oxidized during exponential growth. Our results suggest that prevailing redox conditions constantly control central cellular pathways by fine-tuning oxidation status and hence activity of these proteins. Treatment with sublethal H2O2 concentrations caused a subset of 41 proteins to undergo substantial thiol modifications, thereby affecting a variety of different cellular pathways, many of which are directly or indirectly involved in increasing oxidative stress resistance. Classification of the identified protein thiols according to their steady-state oxidation levels and sensitivity to peroxide treatment revealed that redox sensitivity of protein thiols does not predict peroxide sensitivity. Our studies provide experimental evidence that the ability of protein thiols to react to changing peroxide levels is likely governed by both thermodynamic and kinetic parameters, making predicting thiol modifications challenging and de novo identification of peroxide sensitive protein thiols indispensable.