Edelman William C, Haas Kelsey M, Hsu Joanne I, Lawrence Robert T, Villén Judit
Department of Genome Sciences, University of Washington, Foege S133C, 3720 15th Ave. NE, Seattle, WA, 98105, USA.
Methods Mol Biol. 2014;1156:389-405. doi: 10.1007/978-1-4939-0685-7_26.
The field of cellular signaling is fueled by the discovery of novel protein phosphorylation events. Phosphoproteomics focuses on the large-scale identification and characterization of serine, threonine, and tyrosine phosphorylation of proteins. Phosphopeptide enrichment followed by mass spectrometry has emerged as the most powerful technique for unbiased, discovery-driven analysis by offering high sensitivity, resolution, and speed. Methods for mass spectrometry-based phosphoproteomics analysis have improved substantially over the last decade, making the discipline more approachable to the broader scientific community. Herein we describe the status of the field of phosphoproteomics and provide a robust workflow covering the major aspects of large-scale phosphorylation analysis from phosphopeptide enrichment via IMAC to data analysis.
细胞信号传导领域因新的蛋白质磷酸化事件的发现而蓬勃发展。磷酸化蛋白质组学专注于对蛋白质的丝氨酸、苏氨酸和酪氨酸磷酸化进行大规模鉴定和表征。磷酸肽富集后进行质谱分析已成为最强大的技术，通过提供高灵敏度、分辨率和速度，实现无偏见的、发现驱动的分析。在过去十年中，基于质谱的磷酸化蛋白质组学分析方法有了显著改进，使该学科更容易被广大科学界所接受。在此，我们描述了磷酸化蛋白质组学领域的现状，并提供了一个强大的工作流程，涵盖从通过IMAC进行磷酸肽富集到数据分析的大规模磷酸化分析的主要方面。
