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Methodologies for Characterizing Phosphoproteins by Mass Spectrometry

机译:质谱表征磷酸蛋白的方法

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Posttranslational regulation of proteins via protein phosphorylation is one of the major means of protein regulation. Phosphorylation is a very rapid and reversible method of changing the function of proteins. Detection of phosphorylated proteins and the identification of phosphorylation sites are necessary to molecularly link specific phosphorylated events with change in phosphoprotein function. Mass Spectrometry (MS) has become the methodology of choice for phosphosite identification. Here we review current approaches including sample separation and enrichment techniques (SDS-PAGE, immunoprecipitation, metal-assisted enrichment, strong cation exchange, dendrimer capture), quantitative MS analysis methods (SILAC, iTRAQ, AQUA), and the application of recently developed methods including electron transfer dissociation ionization and “top-down” proteomics to phosphoprotein analysis.
机译:通过蛋白质磷酸化进行蛋白质的翻译后调控是蛋白质调控的主要手段之一。磷酸化是改变蛋白质功能的非常快速且可逆的方法。磷酸化蛋白的检测和磷酸化位点的鉴定对于将特定的磷酸化事件与磷蛋白功能的变化分子连接起来是必需的。质谱(MS)已成为鉴定磷酸位的首选方法。在这里,我们回顾了当前的方法,包括样品分离和富集技术(SDS-PAGE,免疫沉淀,金属辅助富集,强阳离子交换,树枝状聚合物捕获),定量MS分析方法(SILAC,iTRAQ,AQUA)以及最近开发的方法的应用包括电子转移解离电离和“自上而下”的蛋白质组学到磷蛋白分析。

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