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化学进展 2012, Vol. Issue (9): 1785-1793 前一篇   后一篇

• 综述与评论 •

蛋白质组学中磷酸化肽的常用富集方法

李鹏章, 王粤博*   

  1. 山东理工大学化学工程学院 淄博 255049
  • 收稿日期:2011-12-01 修回日期:2012-05-01 出版日期:2012-09-24 发布日期:2012-09-27
  • 通讯作者: 王粤博 E-mail:ybwang@sdut.edu.cn
  • 基金资助:

    山东省中青年科学家科研奖励基金(No.2007BS02003)资助

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Enrichment Methods of Phosphopeptides in Proteomics

Li Pengzhang, Wang Yuebo   

  1. School of Chemical Engineering, Shandong University of Technology, Zibo 255049, China
  • Received:2011-12-01 Revised:2012-05-01 Online:2012-09-24 Published:2012-09-27
磷酸化作用是最重要的蛋白质翻译后修饰方式之一,它是蛋白质组学的一个重要分支,在细胞识别、细胞信息传递、基因表达和新陈代谢等方面发挥着重要作用。采用适当方法对磷酸化肽进行分析有助于我们更好地了解生理病理机制。但是直接进行质谱分析时磷酸化肽的信号强度会受到无机盐以及大量非磷酸化肽的抑制,选择性差。为解决这一难题,在质谱分析前要对磷酸化肽进行选择性富集。本文回顾了几种常用的磷酸化肽富集方法,介绍了每种方法的发展状况和常用材料,其中包括固定金属离子亲和色谱法、金属氧化物富集法、强阴阳离子交换色谱法和MALDI靶板富集法。最后总结了各种富集方法的优缺点,对有效的磷酸化肽富集策略进行了前景展望。
关键词： 磷酸化肽, 固定金属离子亲和色谱, 金属氧化物富集法, 强阳离子交换色谱, 强阴离子交换色谱, MALDI靶板富集法
The reversible phosphorylation of proteins is recognized as an important branch of proteomics. As one of the most important post-translational modifications, protein phosphorylation plays a very important role in cellular recognition, cellular signal transduction, gene expression and cellular metabolism. Exploration of phosphopeptides using appropriate methods can help us further understand the mechanism of physiology and pathology. However, mass spectra (MS) signals of phosphopeptides are frequently suppressed by the presence of abundant nonphosphopeptides and superabundant of salts which will cause poor selectivity. Therefore, selective enrichment of phosphopeptides is essential to facilitate the MS analysis. This paper summarizes several enrichment techniques of phosphopeptides, including immobilized metal affinity chromatography (IMAC), metal oxide enrichment methods, strong cation exchange chromatography (SCX), strong anion exchange chromatography (SAX) and matrix assisted laser desorption ionization (MALDI) on-plate enrichment methods, especially, the materials used in these enrichment methods have been discussed in detail. At the end of this review, the advantages and disadvantages of each method are summarized, and a prospect of effective enrichment strategies of phosphopeptides is given. Contents 1 Introduction
2 Enrichment methods of phosphopeptides
2.1 Immobilized metal affinity chromatography
2.2 Metal oxide enrichment methods
2.3 Strong anion and cation exchange chromatography
2.4 Phosphopeptide enrichment on MALDI plate
3 Conclusion and outlook
Key words: phosphopeptides, immobilized metal affinity chromatography (IMAC), metal oxide enrichment methods, strong cation exchange chromatography (SCX), strong anion exchange chromatography (SAX), MALDI on-plate enrichment methods

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