Development of a Clickable Probe for Profiling of Protein Glutathionylation in the Central Cellular Metabolism of E. coli and Drosophila
Development of a Clickable Probe for Profiling of Protein Glutathionylation in the Central Cellular Metabolism of E. coli and Drosophila
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开发用于分析大肠杆菌和果蝇中央细胞代谢中蛋白质谷胱甘肽化的可点击探针
DOI：
10.1016/j.chembiol.2015.09.012
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发表时间：
2015-11-19
影响因子：
--
通讯作者：
Deng, Haiteng
中科院分区：
文献类型：
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作者：
Feng, Shan;Chen, Yuling;Deng, Haiteng
蛋白质谷胱甘肽化是一种重要的翻译后修饰，可调节许多细胞过程，包括能量代谢、信号转导和蛋白质稳态。谷胱甘肽化蛋白（表示为谷胱甘肽）的全局分析对于理解氧化还原调节的信号转导至关重要。在这里，我们开发了一种基于点击反应和蛋白质组学的新方法来富集和鉴定大肠杆菌和果蝇裂解物中的谷胱甘肽化肽，分别鉴定了937和1,930个潜在的谷胱甘肽化肽。生物信息学分析表明，靠近带负电氨基酸残基的半胱氨酸残基具有更高的谷胱甘肽化频率。重要的是，我们发现大多数与代谢途径相关的蛋白质都被谷胱甘肽化，并且代谢酶的谷胱甘肽化位点在不同物种之间高度保守。我们的研究结果表明，谷胱甘肽类似物是表征蛋白质谷胱甘肽酰化的有用工具，并且在调节细胞代谢中起重要作用的代谢酶的谷胱甘肽酰化是保守的。
Protein glutathionylation is an important post-translational modification that regulates many cellular processes, including energy metabolism, signal transduction, and protein homeostasis. Global profiling of glutathionylated proteins (denoted as glutathionylome) is crucial for understanding redox-regulated signal transduction. Here, we developed a novel method based on click reaction and proteomics to enrich and identify the glutathionylated peptides in Escherichia coli and Drosophila lysates, in which 937 and 1,930 potential glutathionylated peptides were identified, respectively. Bioinformatics analysis showed that the cysteine residue next to negatively charged amino acid residues has a higher frequency of glutathionylation. Importantly, we found that most proteins associated with metabolic pathways were glutathionylated and that the glutathionylation sites of metabolic enzymes were highly conserved among different species. Our results indicate that the glutathione analog is a useful tool to characterize protein glutathionylation, and glutathionylation of metabolic enzymes, which play important roles in regulating cellular metabolism, is conserved.