Department of Cell Biology, Harvard Medical School, Boston, USA.
Department of Genome Sciences, University of Washington, Seattle, USA.
Expert Rev Proteomics. 2021 Sep;18(9):795-807. doi: 10.1080/14789450.2021.1994390. Epub 2021 Oct 28.
Protein phosphorylation is a primary mechanism of signal transduction in cellular systems. Isobaric tagging can be used to investigate alterations in phosphorylation events in sample multiplexing experiments where quantification extends across all conditions. As such, innovations in tandem mass tag methods can facilitate the expansion of the depth and breadth of phosphoproteomic analyses.
This review discusses the current state of tandem mass tag-centric phosphoproteomics and highlights advances in reagent chemistry, instrumentation, data acquisition, and data analysis. We stress that approaches for phosphoproteomic investigations require high-specificity enrichment, sensitive detection, and accurate phosphorylation site localization.
Tandem mass tag-centric phosphoproteomics will continue to be an important conduit for our understanding of signal transduction in living organisms. We anticipate that progress in phosphopeptide enrichment methodologies, enhancements in instrumentation and data acquisition technologies, and further refinements in analytical strategies will be key to the discovery of biologically relevant findings from phosphoproteomics studies.
蛋白质磷酸化是细胞系统信号转导的主要机制。等重标记可用于研究在样品多重实验中磷酸化事件的变化,其中定量可扩展到所有条件。因此,串联质量标签方法的创新可以促进磷酸蛋白质组学分析的深度和广度的扩展。
本文综述了基于串联质谱标签的磷酸蛋白质组学的现状,并重点介绍了试剂化学、仪器、数据采集和数据分析方面的进展。我们强调,磷酸蛋白质组学研究方法需要高特异性富集、敏感检测和准确的磷酸化位点定位。
基于串联质谱标签的磷酸蛋白质组学将继续是我们理解生物体内信号转导的重要途径。我们预计,磷酸肽富集方法学的进展、仪器和数据采集技术的增强以及分析策略的进一步完善,将是从磷酸蛋白质组学研究中发现具有生物学意义的发现的关键。