Selective Targeting of the Cysteine Proteome by Thioredoxin and Glutathione Redox Systems
Selective Targeting of the Cysteine Proteome by Thioredoxin and Glutathione Redox Systems
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DOI：
10.1074/mcp.m113.030437
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发表时间：
2013-11-01
影响因子：
7
通讯作者：
Jones, Dean P.
中科院分区：
文献类型：
--
作者：
Go, Young-Mi;Roede, James R.;Jones, Dean P.
硫氧还蛋白（Trx）和谷胱甘肽（GSH）是保护细胞免受氧化应激诱导的细胞毒性的主要巯基抗氧化剂。Trx和GSH的氧化还原状态已被用作氧化应激的指标。越来越多的研究表明，Trx和GSH氧化还原系统在不同的应激条件下不同地和独立地调节细胞信号传导和代谢途径。在目前的研究中，我们使用了基于质谱的氧化还原蛋白质组学的方法来测试半胱氨酸（Cys）蛋白质组的反应，选择性中断的Trx和GSH依赖的系统。金诺芬（ARF）用于抑制Trx还原酶，而不会检测到GSH/GSSG对的氧化，而丁硫基磺酰亚胺（BSO）用于消耗GSH，而不会检测到Trx 1的氧化。606个含半胱氨酸的肽（肽基半胱氨酸）的结果表明，36%的氧化超过1.3倍的ARF，而BSO诱导的氧化肽基半胱氨酸只有10%。ARF处理的这些肽的平均氧化倍数也高于BSO处理。潜在的功能途径分析表明，ARF氧化肽与糖酵解、细胞骨架重塑、翻译和细胞粘附有关。60肽半胱氨酸氧化由于耗尽的GSH，41也被氧化的ARF和翻译和细胞粘附，但不糖酵解或细胞骨架重塑的蛋白质。测试功能相关性的研究表明，ARF降低了丙酮酸激酶活性和乳酸水平，但BSO没有，证实了对糖酵解相关蛋白的影响对ARF的氧化敏感。这些数据表明，Trx系统比GSH系统调节更广泛的蛋白质，支持Trx和GSH在细胞氧化还原控制中的不同功能，并首次在哺乳动物细胞中显示由于Trx系统功能缺陷而选择性靶向肽基Cys和生物途径。Molecular & Cellular Proteomics 12：10.1074/mcp. M113.030437，3285-3296，2013.
Thioredoxin (Trx) and GSH are the major thiol antioxidants protecting cells from oxidative stress-induced cytotoxicity. Redox states of Trx and GSH have been used as indicators of oxidative stress. Accumulating studies suggest that Trx and GSH redox systems regulate cell signaling and metabolic pathways differently and independently during diverse stressful conditions. In the current study, we used a mass spectrometry-based redox proteomics approach to test responses of the cysteine (Cys) proteome to selective disruption of the Trx- and GSH-dependent systems. Auranofin (ARF) was used to inhibit Trx reductase without detectable oxidation of the GSH/GSSG couple, and buthionine sulfoximine (BSO) was used to deplete GSH without detectable oxidation of Trx1. Results for 606 Cys-containing peptides (peptidyl Cys) showed that 36% were oxidized more than 1.3-fold by ARF, whereas BSO-induced oxidation of peptidyl Cys was only 10%. Mean fold oxidation of these peptides was also higher by ARF than BSO treatment. Analysis of potential functional pathways showed that ARF oxidized peptides associated with glycolysis, cytoskeleton remodeling, translation and cell adhesion. Of 60 peptidyl Cys oxidized due to depletion of GSH, 41 were also oxidized by ARF and included proteins of translation and cell adhesion but not glycolysis or cytoskeletal remodeling. Studies to test functional correlates showed that pyruvate kinase activity and lactate levels were decreased with ARF but not BSO, confirming the effects on glycolysis-associated proteins are sensitive to oxidation by ARF. These data show that the Trx system regulates a broader range of proteins than the GSH system, support distinct function of Trx and GSH in cellular redox control, and show for the first time in mammalian cells selective targeting peptidyl Cys and biological pathways due to deficient function of the Trx system. Molecular & Cellular Proteomics 12: 10.1074/mcp.M113.030437, 3285-3296, 2013.