摘要
cAMP是细胞内一种重要的信号分子,作为第二信使调控包括糖代谢、糖原代谢和脂质代谢在内的一系列生理过程。在本文中我们报道一个新颖的检测细胞内c AMP的基因编码荧光探针,这一探针是由大肠杆菌CAP蛋白和cpYFP荧光蛋白组成的融合蛋白,针对cAMP的结合可以产生一倍的荧光信号响应。进一步的表征实验表明,这一探针对c AMP具有很强的结合能力,并且能够实现快速地荧光响应。这一探针的开发将会为目前已有的检测cAMP的手段提供有益的补充。
cAMP is an important second messenger that is capable of controlling a wide array of cellular processes, including glycogen, sugar and lipid metabolism. Here we report the design and construction of a novel genetically encoded fluorescent sensor for cAMP. The sensor was realized by fusing E. coli CAP protein with cpYFP, and displayed a one-fold fluorescence change towards c AMP binding. Further characterization assays demonstrated that the sensor had high affinity for cAMP and fast response kinetics.The development of our sensor could be a useful supplement to existing methods for cAMP detection.
作者
张嘉远
宋雨欣
王楚宸
王晶
Jiayuan Zhang;Yuxin Song;Chuchen Wang;Jing Wang(State Key Laboratory of Natural and Biomimetic Drugs, Department of Chemical Biology,School of Pharmaceutical Sciences, Peking University Health Science Center, Beijing 100191, China)
基金
National Basic Research Foundation of China(Grant No.2017YFA0505202)
the National Natural Science Foundation of China(Grant No.91853107)
关键词
环腺苷酸
荧光探针
蛋白质工程
信号转导
c AMP
Fluorescent sensor
Protein engineering
Signal transduction
