Analysis of Cysteine Post Translational Modifications Using Organic Mercury Resin.
Analysis of Cysteine Post Translational Modifications Using Organic Mercury Resin.
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DOI：
10.1002/cpps.69
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发表时间：
2018-11
影响因子：
--
通讯作者：
Gould NS
中科院分区：
文献类型：
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作者：
Doulias PT;Gould NS
巯基的广泛反应性使得能够在半胱氨酸残基上形成各种可逆的共价修饰。与许多其他翻译后修饰一样，S-亚硝基化是位点选择性的、可逆的，并且是多种基本细胞过程所必需的。s-亚硝基化蛋白质的总体丰度和亚硝酰基的反应性需要富集策略，以在足够的深度进行准确检测。本文提出了一种利用有机汞树脂从细胞或组织裂解物的复杂混合物中富集和检测内源性蛋白质S-亚硝基化的方法。对该方法的最小调整也支持使用相同的富集平台检测s-谷胱甘肽化或s-酰化。当与高准确度质谱法结合时，这些方法能够实现位点特异性水平的分析，促进三种单独的半胱氨酸翻译后修饰的可比数据集的管理。
The wide reactivity of the thiol group enables the formation of a variety of reversible, covalent modifications on cysteine residues. S-nitrosylation, like many other post-translational modifications, is site selective, reversible, and necessary for a wide variety of fundamental cellular processes. The overall abundance of s-nitrosylated proteins and reactivity of the nitrosyl group necessitates an enrichment strategy for accurate detection with adequate depth. Herein, a method is presented for the enrichment and detection of endogenous protein s-nitrosylation from complex mixtures of cell or tissue lysate utilizing organomercury resin. Minimal adaptations to the method also support the detection of either s-glutathionylation or s-acylation using the same enrichment platform. When coupled with high accuracy mass spectrometry, these methods enable a site specific level of analysis, facilitating the curation comparable datasets of three separate cysteine post-translational modifications.