Quantitative Protein Analysis by Mass Spectrometry
Quantitative Protein Analysis by Mass Spectrometry
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DOI：
10.1007/978-1-4939-2425-7_17
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发表时间：
2015-01-01
期刊：
影响因子：
--
通讯作者：
Peng, Junmin
中科院分区：
文献类型：
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作者：
Pagala, Vishwajeeth R.;High, Anthony A.;Peng, Junmin
质谱学是分析化学中最灵敏的方法之一，在对人类基因组测序后，其在蛋白质组学中的应用迅速扩大。现在，无论是在小范围内还是在整个蛋白质组中，质谱学都是鉴定和定量蛋白质以及翻译后修饰的主流方法。猎枪蛋白质组学可以在一项全面的研究中分析多达10,000种蛋白质，检测灵敏度在皮克范围内。在这一章中，我们描述了一个鸟枪式蛋白质组学平台的主要实验步骤，包括在研究蛋白质-蛋白质相互作用的背景下的样品制备，质谱学数据的获取，以及识别蛋白质的数据库搜索和翻译后修饰分析。还说明了蛋白质组定量策略和生物信息学分析。最后，我们讨论了当前平台的功能、限制和潜在的改进。
Mass spectrometry is one of the most sensitive methods in analytical chemistry, and its application in proteomics has been rapidly expanded after sequencing the human genome. Mass spectrometry is now the mainstream approach for identification and quantification of proteins and posttranslational modifications, either in small scale or in the entire proteome. Shotgun proteomics can analyze up to 10,000 proteins in a comprehensive study, with detection sensitivity in the picogram range. In this chapter, we describe major experimental steps in a shotgun proteomics platform, including sample preparation in the context of studying protein-protein interaction, mass spectrometric data acquisition, and database search to identify proteins and posttranslational modification analysis. Proteome quantification strategies and bioinformatics analysis are also illustrated. Finally, we discuss the capabilities, limitations, and potential improvements of current platforms.