文献类型：期刊文献

英文题名：pH-sensitive pHluorins as a molecular sensor for in situ monitoring of enzyme-catalyzed prodrug activation

作者：Liu, Hui[1];Cao, Xiaodan[1];Wang, Ping[1];Ma, Xingyuan[1]

机构：[1]East China Univ Sci & Technol, Biomed Nanotechnol Ctr, State Key Lab Bioreactor Engn, Shanghai, Peoples R China

年份：2017

卷号：64

期号：4

起止页码：482

外文期刊名：BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY

收录：;EI(收录号：20171703602462);WOS:【SCI-EXPANDED(收录号:WOS:000408126800004)】；

基金：This study was supported by the National Natural Science Foundation (31471659, 31300660, and 1303050), the Science and Technology Innovation Action Plan of Shanghai (14431904300), and the Shanghai Pujiang Rrogram (13PJD012) and was partially supported by the Open Funding Project of the State Key Laboratory of Bioreactor Engineering.

语种：英文

外文关键词：Bcap-37; enzyme; prodrug therapy; HRP; IAA; ratiometric pHluorins

摘要：This work examines the feasibility of using a pH-sensitive fluorescent protein as a molecular reporter for enzyme-catalyzed prodrug activation reaction. Specifically, a ratiometric pHluorins was examined for detection of the activity of horseradish peroxidase (HRP) for the activation of indole-3-acetic acid. The pHluorins and HRP were conjugated chemically, forming a biocatalyst with a self-reporting function. Results showed that the characteristic fluorescence intensity ratio of the conjugate shifted from 1.47 to 1.40 corresponding to the progress of the prodrug activation reaction. The effectiveness of applying the conjugate for inhibition of the growth of Bcap-37 cells was also demonstrated simultaneously with reaction monitoring. The results reveal a very promising approach to realizing in situ monitoring of enzyme activities based on pH shifting for enzyme-based prodrug therapy applications. (C) 2016 International Union of Biochemistry and Molecular Biology, Inc.