文献类型：期刊文献

英文题名：Enzymatic Cleavage and Subsequent Facile Intramolecular Transcyclization for in Situ Fluorescence Detection of γ-Glutamyltranspetidase Activities

作者：Tong, Hongjuan[1,2];Zheng, Yongjun[1,2];Zhou, Li[1,2];Li, Xiangmin[1,2];Qian, Rui[1,2];Wang, Rui[1,2];Zhao, Jianhong[1,2];Lou, Kaiyan[1,2];Wang, Wei[1,2,3]

机构：[1]East China Univ Sci & Technol, Sch Pharm, Shanghai Key Lab Chem Biol, Shanghai Key Lab New Drug Design, 130 Meilong Rd, Shanghai 200237, Peoples R China;[2]East China Univ Sci & Technol, State Key Lab Bioengn Reactor, 130 Meilong Rd, Shanghai 200237, Peoples R China;[3]Univ New Mexico, Dept Chem & Chem Biol, Albuquerque, NM 87131 USA

年份：2016

卷号：88

期号：22

起止页码：10816

外文期刊名：ANALYTICAL CHEMISTRY

收录：;EI(收录号：20181705037112);WOS:【SCI-EXPANDED(收录号:WOS:000388154700008)】；

基金：The work was supported by East China University of Science and Technology (start-up funds, W.W.), the Fundamental Research Funds for the Central Universities (Grant WY1213013, K. L.), the Pujiang Talent Project (Grant 14PJ1402200, K. L.), the National Science Foundation of China (Grant No. 21577037, K. L.), and the China 111 Project (Grant B07023, W.W.).

语种：英文

外文关键词：Cancer cells - Cells - Diseases - Diagnosis - Cytology - Fluorescence spectroscopy - Probes - Enzymes - Product design

摘要：gamma-Glutamyltranspetidase (GGT) is a cell-membrane-bound enzyme which selectively catalyzes cleavage of the gamma-glutamyl bond of glutathione (GSH). It has been identified to be overexpressed in a number of malignant tumor cells. Therefore, fluorescent probes for fast and selective detection of GGT activities are greatly needed. However, the majority of currently available GGT fluorescent probes based on direct conjugation of a gamma-glutamyl group to a specific fluorophore generally has slow enzymatic kinetics due to bulky fluorophore too close to the enzyme's active site. Moreover, the uncaged fluorophore with a free amine group might undergo oxidation or other enzymatic transformation and resulted in a complicated time dependent fluorescence response. Herein, we reported design of a novel fluorescent GGT probe NM-GSH (2), which incorporated a fast intramolecular transcyclization cascade for rapid detection of GGT activities after enzymatic cleavage of the gamma-glutamyl group. This design strategy allows introduction of bulky 1,8-naphthalimide fluorophore with improved enzymatic kinetics and lowered detection limit. The transcyclized product 4 gives more than 200-fold fluorescence increment. The probe NM-GSH showed both good selectivity and fast detection of GGT activities with the detection limit as low as 0.21 mU/mL. In addition, the fluorescent product 4 contains no free amine group and is more stable for detection. Most importantly, cell imaging studies showed that the transcyclized product 4 was enriched in lysosomes for selectively lighting up GGT-overexpressed ovarian cancer cells (OVCARS) but not normal cells (HUVEC), indicating NM-GSH's potentials as an imaging agent in cancer diagnosis and treatment.