Department of Chemistry and Purdue University Center for Cancer Research, Purdue University, 560 Oval Drive, West Lafayette, IN 47907, USA.
IUBMB Life. 2019 Jun;71(6):721-737. doi: 10.1002/iub.2023. Epub 2019 Feb 22.
Protein kinases function as key signaling hubs in the intricate network of biochemical signaling processes in the living cell. More than two-thirds of the human proteome is estimated to be phosphorylated at ~960,000 phosphosites, which makes it challenging to identify the direct contribution of any desired kinase in generating this phosphoproteome. In this review, we discuss some of the methods that have been developed over the years for global identification of kinase substrates. The methods are essentially categorized into two classes, namely, (i) direct tagging of kinase substrates and (ii) indirect phosphoproteomics-based approaches. We discuss the advantages and limitations entailed to each of the method introduced, with a special emphasis on the analog-sensitive (as) kinase approach method. © 2019 IUBMB Life, 71(6):721-737, 2019.
蛋白质激酶作为关键的信号枢纽，存在于活细胞内复杂的生化信号转导网络中。据估计，人类蛋白质组中超过三分之二的蛋白质在约 96 万个磷酸化位点被磷酸化，这使得确定任何所需激酶在产生这种磷酸化蛋白质组中的直接贡献具有挑战性。在这篇综述中，我们讨论了多年来为全球鉴定激酶底物而开发的一些方法。这些方法基本上分为两类，即：（i）激酶底物的直接标记和（ii）基于间接磷酸蛋白质组学的方法。我们讨论了每种方法的优缺点，特别强调了模拟敏感(as)激酶方法。国际生物化学与分子生物学联合会生命杂志，2019 年 71(6)：721-737.
