Department of Chemistry, Durham University, Science Laboratories, Durham DH1 3LE, United Kingdom.
Chem Soc Rev. 2011 Mar;40(3):1211-23. doi: 10.1039/c0cs00020e. Epub 2010 Dec 13.
Protein kinases control the function of about one third of cellular proteins by catalysing the transfer of the γ-phosphate group of ATP onto their substrate proteins. Protein phosphatases counter this action and also control the activation status of many kinases. Cellular responses to environmental changes, or signalling events, temporarily tilt the balance of protein phosphorylation and dephosphorylation to one side or the other. The identification of protein-kinase-substrate pairs and substrate-phosphatase pairs is critical to understanding cell function and how cells respond to environmental changes. Identification of these substrate-enzyme pairs is non-trivial, because of the structural and mechanistic conservation of the catalytic cores of protein kinases. In this tutorial review we review recent progress towards identifying protein-kinase-substrate pairs by emphasising the use of chemical genetics and purpose-designed ATP analogues that target one particular protein kinase. In addition, we discuss activity-based chemical profiling approaches, based on ATP analogues, for the detection of active kinases.
蛋白激酶通过催化 ATP 的γ-磷酸基团转移到其底物蛋白上，控制大约三分之一的细胞蛋白的功能。蛋白磷酸酶则拮抗这种作用，并控制许多激酶的激活状态。细胞对环境变化或信号事件的反应会暂时使蛋白磷酸化和去磷酸化的平衡向一侧倾斜。鉴定蛋白激酶-底物对和底物-磷酸酶对对于理解细胞功能以及细胞如何对环境变化作出反应至关重要。由于蛋白激酶催化核心的结构和机制保守，这些底物-酶对的鉴定并非易事。在本综述中，我们强调了化学遗传学和专门设计的靶向特定蛋白激酶的 ATP 类似物的应用，介绍了鉴定蛋白激酶-底物对的最新进展。此外，我们还讨论了基于 ATP 类似物的基于活性的化学特征分析方法，用于检测活性激酶。
