Ma Changbei, Liu Haisheng, Du Junyan, Chen Hanchun, He Hailun, Jin Shunxin, Wang Kemin, Wang Jun
State Key Laboratory of Medical Genetics & School of Life Sciences, Central South University, Changsha 410013, China; State Key Laboratory of Chemo/Biosensing and Chemometrics, Hunan University, Changsha 410081, China.
State Key Laboratory of Medical Genetics & School of Life Sciences, Central South University, Changsha 410013, China.
Anal Biochem. 2016 Feb 1;494:1-3. doi: 10.1016/j.ab.2015.10.007. Epub 2015 Oct 28.
Traditional methods of assaying polynucleotide kinase (PNK) activity are discontinuous, time-consuming, and laborious. Here we report a new quencher-free approach to real-time monitoring of PNK activity using a 2-aminopurine probe. When the 2-aminopurine probe was 5'-phosphorylated by PNK, it could be efficiently degraded by lambda exonuclease to release free 2-aminopurine molecules and generate a fluorescence signal. This method not only provides a universal approach to real-time monitoring of PNK activity, but also shows great potential for screening suitable inhibitor drugs for PNK.
传统的多核苷酸激酶（PNK）活性检测方法是不连续的，耗时且费力。在此，我们报告一种使用2-氨基嘌呤探针实时监测PNK活性的无淬灭剂新方法。当2-氨基嘌呤探针被PNK 5'-磷酸化时，它可被λ核酸外切酶有效降解，释放出游离的2-氨基嘌呤分子并产生荧光信号。该方法不仅提供了一种实时监测PNK活性的通用方法，而且在筛选适合PNK的抑制剂药物方面也显示出巨大潜力。
