质谱技术分析磷酸化蛋白的研究进展

Advances in the Analysis of Phosphorylated Protein by Mass Spectrometry

蛋白质的可逆磷酸化是最重要的翻译后修饰之一,几乎参与生命活动的所有过程。虽然对于磷酸化蛋白的全面解析还存在巨大挑战,质谱已逐渐被人们认为是挑战这一领域的有利工具。质谱技术分析磷酸化蛋白质的方法包括利用抗体免疫沉淀、磷酸化蛋白胶染色、固定化的金属亲和层析柱、化学标签、强阳离子交换等技术富集磷酸化蛋白或肽段,串联质谱、电子俘获分析等技术检测磷酸化肽段并鉴定磷酸化位点,以及同位素标签对蛋白质磷酸化水平进行定量等。

Reversible phosphorylation of proteins is one of the most important post-translational modifications and is also involved in a host of cellular processes. Although full analysis of phosphorylated proteins remains a formidable analytical challenge, mass spectrometry has emerged as an increasingly viable tool for this task. This review summarizes the methodologies currently available for the analysis of phosphorylated proteins by mass spectrometry including immunoprecipitating proteins using antibodies against phosphorylated residues, fluorescent phosphoprotein gel staining, immobilized metal affinity chromatography, chemical modification, strong cation exchange to enrich the phosphoproteins or phosphopeptides. Tandem mass spectrometry approach and electron capture dissociation have been applied for the identification of phosphorylation and analysis of phosphorylation sites. Finally, isotopic labeling are proposed for the the quantification of phosphorylation sites.