Rosenqvist Heidi, Ye Juanying, Jensen Ole N
Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow G4 0RE, Scotland, UK.
Methods Mol Biol. 2011;753:183-213. doi: 10.1007/978-1-61779-148-2_13.
Phosphoproteomics, the systematic study of protein phosphorylation events and cell signaling networks in cells and tissues, is a rapidly evolving branch of functional proteomics. Current phosphoproteomics research provides a large toolbox of strategies and protocols that may assist researchers to reveal key regulatory events and phosphorylation-mediated processes in the cell and in whole organisms. We present an overview of sensitive and robust analytical methods for phosphopeptide analysis, including calcium phosphate precipitation and affinity enrichment methods such as IMAC and TiO(2). We then discuss various tandem mass spectrometry approaches for phosphopeptide sequencing and quantification, and we consider aspects of phosphoproteome data analysis and interpretation. Efficient integration of these stages of phosphoproteome analysis is highly important to ensure a successful outcome of large-scale experiments for studies of phosphorylation-mediated protein regulation.
磷酸化蛋白质组学是对细胞和组织中蛋白质磷酸化事件及细胞信号网络进行的系统研究，是功能蛋白质组学中一个快速发展的分支。当前的磷酸化蛋白质组学研究提供了大量的策略和方案工具箱，可帮助研究人员揭示细胞和整个生物体中的关键调控事件以及磷酸化介导的过程。我们概述了用于磷酸肽分析的灵敏且可靠的分析方法，包括磷酸钙沉淀法以及诸如IMAC和TiO(2)等亲和富集方法。然后我们讨论了用于磷酸肽测序和定量的各种串联质谱方法，并考虑了磷酸化蛋白质组数据分析和解释的各个方面。磷酸化蛋白质组分析这些阶段的有效整合对于确保大规模实验成功研究磷酸化介导的蛋白质调控至关重要。
