Discovery of a butyrylcholinesterase-specific probe via a structure-based design strategy.
We report herein the structure-based design and application of a fluorogenic molecular probe (BChE-FP) specific to butyrylcholinesterase (BChE). This probe was rationally designed by mimicking the native substrate and optimized stepwise by manipulating the steric feature and the reactivity of the designed probe targeting the structural difference of the active pockets of BChE and AChE. The refined probe, BChE-FP, exhibits high specificity toward BChE compared to AChE, producing about 275-fold greater fluorescence enhancement upon the catalysis by BChE. Thus, BChE-FP is a specific BChE probe identified by the structure-based design and it can discriminate BChE from AChE. Furthermore, it has been successfully applied for imaging the endogenous BChE in living cells, as well as BChE inhibitor screening and characterization under physiological conditions.
Key Laboratory of Pesticide and Chemical Biology, Ministry of Education, College of Chemistry, Central China Normal University, Wuhan 430079, P. R. China. tomyang@mail.ccnu.edu.cn gfyang@mail.ccnu.edu.cn.
Shu-Hou
SH
Yang
Key Laboratory of Pesticide and Chemical Biology, Ministry of Education, College of Chemistry, Central China Normal University, Wuhan 430079, P. R. China. tomyang@mail.ccnu.edu.cn gfyang@mail.ccnu.edu.cn.
Qi
Q
Sun
Key Laboratory of Pesticide and Chemical Biology, Ministry of Education, College of Chemistry, Central China Normal University, Wuhan 430079, P. R. China. tomyang@mail.ccnu.edu.cn gfyang@mail.ccnu.edu.cn.
Hao
H
Xiong
Key Laboratory of Pesticide and Chemical Biology, Ministry of Education, College of Chemistry, Central China Normal University, Wuhan 430079, P. R. China. tomyang@mail.ccnu.edu.cn gfyang@mail.ccnu.edu.cn.
Shi-Yu
SY
Liu
Key Laboratory of Pesticide and Chemical Biology, Ministry of Education, College of Chemistry, Central China Normal University, Wuhan 430079, P. R. China. tomyang@mail.ccnu.edu.cn gfyang@mail.ccnu.edu.cn.
Behrooz
B
Moosavi
Key Laboratory of Pesticide and Chemical Biology, Ministry of Education, College of Chemistry, Central China Normal University, Wuhan 430079, P. R. China. tomyang@mail.ccnu.edu.cn gfyang@mail.ccnu.edu.cn.
Wen-Chao
http://orcid.org/0000-0002-6722-0441
WC
Yang
Key Laboratory of Pesticide and Chemical Biology, Ministry of Education, College of Chemistry, Central China Normal University, Wuhan 430079, P. R. China. tomyang@mail.ccnu.edu.cn gfyang@mail.ccnu.edu.cn and Collaborative Innovation Center of Chemical Science and Engineering, Tianjin 30071, P. R. China.
Guang-Fu
GF
Yang
D000110:Acetylcholinesterase / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D002091:Butyrylcholinesterase / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D045744:Cell Line, Tumor; D055808:Drug Discovery; D005456:Fluorescent Dyes / Q000032:analysis* / Q000138:chemical synthesis* / Q000737:chemistry; D006801:Humans; D008958:Models, Molecular; D015394:Molecular Structure; D061848:Optical Imaging
D005456:Fluorescent Dyes; D000110:Acetylcholinesterase; D002091:Butyrylcholinesterase