Quantitative Analysis of in Vivo Methionine Oxidation of the Human Proteome
Quantitative Analysis of in Vivo Methionine Oxidation of the Human Proteome
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DOI：
10.1021/acs.jproteome.9b00505
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发表时间：
2020-02-01
影响因子：
4.4
通讯作者：
Ghaemmaghami, Sina
中科院分区：
文献类型：
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作者：
Bettinger, John Q.;Welle, Kevin A.;Ghaemmaghami, Sina
蛋氨酸的氧化是一种重要的蛋白质翻译后修饰，在生理学和病理学中具有许多作用。然而，在蛋白质组范围内定量分析蛋氨酸氧化受到技术限制。在样品制备和分析过程中，甲基苯丙胺很容易在体外氧化。此外，缺乏含有氧化甲硫氨酸残基的肽的富集方案，使得难以在全球范围内实现甲硫氨酸氧化的准确定量。在这里，我们报告了一种方法来规避这些问题，同位素标记未氧化的蛋氨酸与O-18-标记的过氧化氢和定量的O-18-和O-16-氧化蛋氨酸的相对比例。我们使用人工氧化的蛋白质组来模拟不同程度的蛋氨酸氧化，从而验证我们的方法。使用这种方法，我们确定和量化了一些新的网站在体内蛋氨酸氧化的非应激人类细胞系。
The oxidation of methionine is an important post-translational modification of proteins with numerous roles in physiology and pathology. However, the quantitative analysis of methionine oxidation on a proteome-wide scale has been hampered by technical limitations. Methionine is readily oxidized in vitro during sample preparation and analysis. In addition, there is a lack of enrichment protocols for peptides that contain an oxidized methionine residue, making the accurate quantification of methionine oxidation difficult to achieve on a global scale. Herein, we report a methodology to circumvent these issues by isotopically labeling unoxidized methionines with O-18-labeled hydrogen peroxide and quantifying the relative ratios of O-18- and O-16-oxidized methionines. We validate our methodology using artificially oxidized proteomes made to mimic varying degrees of methionine oxidation. Using this method, we identify and quantify a number of novel sites of in vivo methionine oxidation in an unstressed human cell line.