文献类型：期刊文献

中文题名：High-contrast and real-time visualization of membrane proteins in live cells with malachite green-based fluorogenic probes

作者：Yefeng Chen[1];Chenghong Xue[1];Jie Wang[1];Minqiu Xu[1];Yuyao Li[1];Yiru Ding[1];Heng Song[1];Weipan Xu[1];Hexin Xie[1]

机构：[1]State Key Laboratory of Bioreactor Engineering,Shanghai Key Laboratory of New Drug Design,Frontiers Science Center for Materiobiology and Dynamic Chemistry,School of Pharmacy,East China University of Science and Technology,Shanghai 200237,China

年份：2022

卷号：33

期号：3

起止页码：1637

中文期刊名：Chinese Chemical Letters

外文期刊名：中国化学快报（英文版）

收录：CSTPCD;;Scopus;CSCD:【CSCD2021_2022】；PubMed;

基金：financially supported by the National Natural Science Foundation of China (No. 22077031);the Research Program of State Key Laboratory of Bioreactor Engineering;the Fundamental Research Funds for the Central Universities。

语种：英文

中文关键词：Cellular imaging;Fluorogenic probe;Environment-sensitive fluorophore;Alkaline phosphatase;Integrin

摘要：Imaging dynamics of membrane proteins of live cells in a wash-free and real-time manner has been a challenging task. Herein, we report unprecedented applications of malachite green(MG), an organic dye widely used in pigment industry, as a switchable fluorophore to monitor membrane enzymes or noncatalytic proteins in live cells. Conformationally flexible MG is non-fluorescent in aqueous solution, yet covalent binding with endogenous proteins of cells significantly enhances its fluorescence at 670 nm by restricting flexibility of dye. Integrating a phosphate-caged quinone methide precursor with MG yielded a covalent labeling fluorogenic probe, allowing real-time imaging of membrane alkaline phosphatase(ALP,a model catalytic protein) activity in live cells with over 100-fold enhancement of fluorescence intensity.Moreover, MG is also applicable to image non-catalytic protein by conjugation with protein-specific ligand. A fluorogenic probe consisted of c-RGDf K peptide and MG proved to be compatible with wash-free and real-time visualization of non-catalytic integrin α_(v)β_(3) in live cells with high contrast.