Harvard Medical School, Boston, Massachusetts, USA.
University of Washington, Seattle, Washington, USA.
Proteomics. 2021 May;21(9):e2000140. doi: 10.1002/pmic.202000140. Epub 2021 Mar 30.
Eukaryotic protein phosphorylation modulates nearly every major biological process. Phosphorylation regulates protein activity, mediates cellular signal transduction, and manipulates cellular structure. Consequently, the dysregulation of kinase and phosphatase pathways has been linked to a multitude of diseases. Mass spectrometry-based proteomic techniques are increasingly used for the global interrogation of perturbations in phosphorylation-based cellular signaling. Strategies for studying phosphoproteomes require high-specificity enrichment, sensitive detection, and accurate localization of phosphorylation sites with advanced LC-MS/MS techniques and downstream informatics. Sample multiplexing with isobaric tags has also been integral to recent advancements in throughput and sensitivity for phosphoproteomic studies. Each of these facets of phosphoproteomics analysis present distinct challenges and thus opportunities for improvement and innovation. Here, we review current methodologies, explore persistent challenges, and discuss the outlook for isobaric tag-based quantitative phosphoproteomic analysis.
真核生物蛋白磷酸化调节着几乎每一个主要的生物过程。磷酸化调节蛋白质活性、介导细胞信号转导、并操纵细胞结构。因此，激酶和磷酸酶途径的失调与多种疾病有关。基于质谱的蛋白质组学技术越来越多地用于对基于磷酸化的细胞信号转导的扰动进行全局研究。研究磷酸蛋白质组学的策略需要高特异性的富集、灵敏的检测和准确的磷酸化位点定位，这需要先进的 LC-MS/MS 技术和下游信息学。利用等重标记物进行样品多重化也是提高磷酸蛋白质组学研究通量和灵敏度的重要手段。磷酸蛋白质组学分析的这些方面都存在着独特的挑战，因此也为改进和创新提供了机会。在这里，我们回顾了现有的方法，探讨了存在的挑战，并讨论了基于等重标记物的定量磷酸蛋白质组学分析的前景。
