Sensing cytochrome P4501A1 activity by a resorufin-based isoform-specific fluorescent probe  

作　　者：Qiang Jin Hongying Ma Lei Feng Ping Wang Rongjing He Jing Ning Ling Yang Guangbo Ge 

机构地区：[1]Institute of Interdisciplinary Integrative Medicine Research,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China [2]College of Pharmacy,Dalian Medical University,Dalian 116044,China

出　　处：《Chinese Chemical Letters》2020年第11期2945-2949,共5页中国化学快报（英文版）

基　　金：the National Natural Science Foundation of China(Nos.81922070,81973286,81803489,81773687 and 81703604);the National Key Research and Development Program of China(Nos.2016YFC1303900,2017YFC1700200 and 2017YFC1702000);the Three-year Action Plan of Shanghai TCM Development(No.ZY-(2018-2020)-CCCX5001);Drug Innovation Major Project(No.2018ZX09731016);Program of Shanghai Academic/Technology Research Leader(No.18XD1403600);Shuguang Program(No.18SG40)supported by Shanghai Education Development Foundation and Shanghai Municipal Education Commission。

摘　　要：Cytochrome P4501 A1(CYP1A1),a heme-containing monooxygenase,is of particular importance for human health because of its vital roles in the metabolic activation of pro-carcinogenic compounds to the carcinogens.Deciphering the relevance of CYP1A1 to human diseases and screening of CYP1A1 modulators require reliable tool(s)for probing this key enzyme in complex biological matrices.Herein,a practical and ultrasensitive fluorescence-based assay for real-time sensing CYP1A1 activities in biological systems has been developed,via designing an isoform-specific fluorogenic sensor for CYP1A1(CHPO).The newly developed fluorogenic substrate for CYP1A1 has been carefully investigated in terms of specificity,sensitivity,precision,quantitative linear range and the anti-interference ability.The excellent selectivity,strong anti-interference ability and fast response kinetics,making the practicability of CHPObased CYP1A1 activity assay is better than that of most reported CYP1A1 activity assays.Furthermore,CHPO has been successfully used for imaging CYP1A1 activities in living cells and human tissues,as well as for high-throughput screening of CYP1A1 inhibitors using tissue preparations as enzyme sources.Collectively,this study provided a practical fluorogenic sensor for real-time sensing CYP1A1 in complex biological systems,which would strongly facilitate the investigations on the relevance of CYP1A1 to human diseases and promote high-throughput screening of CYP1A1 modulators for biomedical applications.

关 键 词：Cytochrome P4501A1 Fluorogenic sensor Activity sensing RESORUFIN Isoform-specificity BIOIMAGING 

分 类 号：R318[医药卫生—生物医学工程] O657.3[医药卫生—基础医学]