Tsang Tsz Fung, Qiu Yangyi, Lin Lin, Ye Jiqing, Ma Cong, Yang Xiao
Department of Microbiology , The Chinese University of Hong Kong, Prince of Wales Hospital , 30-32 Ngan Shing Street , Shatin , Hong Kong.
The Hong Kong Polytechnic University Shenzhen Research Institute, 18 Yue Xing 1st Road , Nanshan, Shenzhen , Guangdong 518057 , China.
ACS Infect Dis. 2019 Apr 12;5(4):521-527. doi: 10.1021/acsinfecdis.9b00020. Epub 2019 Mar 6.
Protein-protein interactions (PPIs) underpin essential cellular processes of all organisms and are increasingly considered as drug targets. A number of techniques have been established to study PPIs; however, development of a simple and cost-effective method for in vitro high throughput screening of PPI inhibitors is still in demand or desirable. We report herein a simple method based on protein complementation for the in vitro study of PPIs, as well as screening of inhibitors against the PPI of interest. We have validated this system utilizing bacterial transcription factors NusB and NusE. Three derivatives of an inhibitor targeting the NusB-NusE interaction were synthesized and characterized with the system, which showed specific inhibition and antimicrobial activities. We have further confirmed the system with the RNA polymerase-σ interaction and an inhibitor. This system is expected to be suitable for more extensive high throughput screening of large chemical libraries. Additionally, our vector system can be easily adapted to study other PPI pairs, followed by inhibitor screening for hit identification in the application of early stage drug discovery.
蛋白质-蛋白质相互作用（PPIs）是所有生物体基本细胞过程的基础，并且越来越被视为药物靶点。已经建立了许多技术来研究蛋白质-蛋白质相互作用；然而，开发一种用于体外高通量筛选蛋白质-蛋白质相互作用抑制剂的简单且经济高效的方法仍然是有需求的或令人期待的。我们在此报告一种基于蛋白质互补的简单方法，用于蛋白质-蛋白质相互作用的体外研究以及针对感兴趣的蛋白质-蛋白质相互作用筛选抑制剂。我们利用细菌转录因子NusB和NusE验证了该系统。合成了三种靶向NusB-NusE相互作用的抑制剂衍生物，并通过该系统进行了表征，结果显示出特异性抑制作用和抗菌活性。我们进一步用RNA聚合酶-σ相互作用及一种抑制剂对该系统进行了验证。该系统有望适用于对大型化学文库进行更广泛的高通量筛选。此外，我们的载体系统可以很容易地用于研究其他蛋白质-蛋白质相互作用对，随后在早期药物发现应用中进行抑制剂筛选以鉴定活性化合物。
