Xuan Teng-Fei, Liu Jun, Wang Zi-Qiang, Chen Wei-Min, Lin Jing
International Cooperative Laboratory of Traditional Chinese Medicine Modernization and Innovative Drug Development of Chinese Ministry of Education (MOE), College of Pharmacy, Jinan University, Guangzhou, China.
Front Microbiol. 2020 Jan 14;10:3163. doi: 10.3389/fmicb.2019.03163. eCollection 2019.
3',5' Cyclic diguanylic acid (c-di-GMP) has been shown to play a central role in the regulation of bacterial physiological processes such as biofilm formation and virulence production, and is regarded as a potential target for the development of anti-infective drugs. A method for the facile detection of the bacterial level of cellular c-di-GMP is required to explore the details of c-di-GMP signaling and design drugs on the basis of this pathway. Current methods of c-di-GMP detection have limited sensitivity or difficultly in probe preparation. Herein a new fluorescent probe is reported for the detection of c-di-GMP at concentrations as low as 500 nM. The probe was developed on the basis of the G-quadruplex formation of c-di-GMP induced by aromatic molecules. When used on crude bacterial cell lysates, it can effectively distinguish between the low c-di-GMP levels of bacteria in plankton and the high c-di-GMP levels in biofilm. The method described here is simple, inexpensive, sensitive, and suitable for practical applications involving the rapid detection of cellular c-di-GMP levels after simple bacterial lysis and filtration.
3',5'-环二鸟苷酸（c-di-GMP）已被证明在调节细菌生理过程（如生物膜形成和毒力产生）中起核心作用，并且被视为抗感染药物开发的潜在靶点。需要一种简便的方法来检测细菌细胞内c-di-GMP的水平，以探究c-di-GMP信号传导的细节并基于此途径设计药物。目前检测c-di-GMP的方法灵敏度有限或探针制备困难。本文报道了一种新型荧光探针，可检测低至500 nM浓度的c-di-GMP。该探针是基于芳香分子诱导c-di-GMP形成G-四链体而开发的。当用于粗制细菌细胞裂解物时，它可以有效区分浮游细菌中低水平的c-di-GMP和生物膜中高水平的c-di-GMP。本文所述方法简单、廉价、灵敏，适用于在简单细菌裂解和过滤后快速检测细胞内c-di-GMP水平的实际应用。
