蛋白激酶的生化活性检测方法研究进展

The progress of biochemical activity assays against protein kinases

蛋白激酶在真核细胞信号转导中起重要作用,影响了生长、发育、迁移以及凋亡等各个细胞过程。其表达水平或活性异常时,就有可能导致癌症、心血管疾病以及其他各种疾病,因此蛋白激酶是治疗这些疾病很好的分子靶点。迄今为止,美国食品药品监督管理局已经批准了28个蛋白激酶的抑制剂作为上市药物,用于相应的临床治疗。目前存在着各种检测激酶活性的方法,激酶生化检测方法尤为众多,比较经典的有放射性同位素的方法,也有一些非均相非放射性同位素的方法,诸如酶联免疫法、反相高效液相色谱法、核磁共振分析法等等。而各种均相非放射性同位素的检测方法,由于其污染小、操作便捷,逐渐成为激酶抑制剂筛选的首选。本文综述了各种激酶生化检测方法及其发展历史,并介绍了一些新的趋势,如激酶酶谱筛选。

Protein kinases play critical roles in signal transduction in eukaryotic cells, such as cell growth, development, migration and apoptosis. When their expression levels or activities are abnormal, it possibly leads to many diseases, such as cancer, cardiovascular disease and so on. So protein kinases are good mo- lecular targets to cure related diseases. Up to now, 28 protein kinases inhibitors have been approved by US Food and Drug Administration （FDA） as drugs in the clinical treatment market. Many kinase activity assays, especially biochemical activity assays have been developed to screen novel kinase inhibitors. The principle of the most classical kinase assays is utilizing radioactivity reagents for detection. Meanwhile, some non-ra- dioactivity and heterogeneous assays, such as enzyme-linked immunosorbent assay （ELISA）, reversed-phase high performance liquid chromatography （RP-HPLC）, nuclear magnetic resonance （NMR） have been devel- oped. Furthermore, the non-radioactivity homogeneous assays become predominant in the kinase screening field because of low contamination and convenient operational procedure. This article summarized the kinase biochemical assays in the market and the history of kinase screening methods. Meanwhile, some new trends, such as kinase profiling are introduced.