Cancer Research Institute, Biomedical Research Center, Slovak Academy of Sciences, Dubravska cesta 9, 845 05 Bratislava, Slovakia.
Institute of Chemistry, Slovak Academy of Sciences, Dubravska cesta 9, 845 38 Bratislava, Slovakia.
Int J Mol Sci. 2020 Oct 15;21(20):7637. doi: 10.3390/ijms21207637.
Protein kinases are important enzymes involved in the regulation of various cellular processes. To function properly, each protein kinase phosphorylates only a limited number of proteins among the thousands present in the cell. This provides a rapid and dynamic regulatory mechanism that controls biological functions of the proteins. Despite the importance of protein kinases, most of their substrates remain unknown. Recently, the advances in the fields of protein engineering, chemical genetics, and mass spectrometry have boosted studies on identification of bona fide substrates of protein kinases. Among the various methods in protein kinase specific substrate identification, genetically engineered protein kinases and quantitative phosphoproteomics have become promising tools. Herein, we review the current advances in the field of chemical genetics in analog-sensitive protein kinase mutants and highlight selected strategies for identifying protein kinase substrates and studying the dynamic nature of protein phosphorylation.
蛋白激酶是参与调节各种细胞过程的重要酶。为了正常发挥功能，每种蛋白激酶仅能使细胞中存在的数千种蛋白质中的有限数量的蛋白质发生磷酸化。这提供了一种快速而动态的调节机制，可控制蛋白质的生物学功能。尽管蛋白激酶非常重要，但它们的大多数底物仍然未知。最近，在蛋白质工程、化学遗传学和质谱学领域的进展推动了对蛋白激酶真正底物的鉴定研究。在蛋白激酶特异性底物鉴定的各种方法中，基因工程蛋白激酶和定量磷酸化蛋白质组学已成为有前途的工具。本文综述了化学遗传学领域中模拟敏感蛋白激酶突变体的最新进展，并重点介绍了鉴定蛋白激酶底物和研究蛋白磷酸化动态性质的几种策略。
