文献类型：期刊文献

英文题名：Selective and Ratiometric Fluorescent Trapping and Quantification of Protein Vicinal Dithiols and in Situ Dynamic Tracing in Living Cells

作者：Huang, Chusen[1,2,3];Jia, Ti[2,3];Tang, Mengfang[1];Yin, Qin[1];Zhu, Weiping[1];Zhang, Chao[1];Yang, Yi[1];Jia, Nengqin[2,3];Xu, Yufang[1];Qian, Xuhong[1]

机构：[1]E China Univ Sci & Technol, Shanghai Key Lab Chem Biol, Sch Pharm, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]Shanghai Normal Univ, Educ Minist, Key Lab Resource Chem, Shanghai 200234, Peoples R China;[3]Shanghai Normal Univ, Shanghai Key Lab Rare Earth Funct Mat, Dept Chem, Coll Life & Environm Sci, Shanghai 200234, Peoples R China

年份：2014

卷号：136

期号：40

起止页码：14237

外文期刊名：JOURNAL OF THE AMERICAN CHEMICAL SOCIETY

收录：;EI(收录号：20144300120620);WOS:【SCI-EXPANDED(收录号:WOS:000343026700044)】；

基金：We thank the financial support from National Natural Science Foundation of China (Grants 21476077, 21236002, 21373138, 21302125), National Basic Research Program of China (973 Program, 2013CB733700, 2010CB126100), National High Technology Research and Development Program of China (863 Program, 2011AA10A207), Shanghai Pujiang Program, Doctoral Fund of Ministry of Education of China (Grant No. 20133127120005), Program for Shanghai Sci. & Tech. Committee (Grants 13ZR1458800), Fundamental Research Funds for the Central Universities, Program for Cultivation of Young Teacher of Shanghai University.

语种：英文

外文关键词：Sulfur compounds - Cell membranes - Fluorescence imaging - Biosynthesis - Probes - Cytology

摘要：Protein vicinal dithiols play fundamental roles in intracellular redox homeostasis due to their involvement in protein synthesis and function through the reversible vicinal dithiol oxidation to disulfide. To provide quantitative information about the global distribution and dynamic changes of protein vicinal dithiols in living cells, we have designed and synthesized a ratiometric fluorescent probe (VTAF) for trapping of vicinal dithiol-containing proteins (VDPs) in living cells. VTAF exhibits a ratiometric fluorescence signal upon single excitation, which enables self-calibration of the fluorescence signal and quantification of endogenous vicinal dithiols of VDPs. Its potential for in situ dynamic tracing of changes of protein vicinal dithiols under different cellular redox conditions was exemplified. VTAF facilitated the direct observation of subcellular distribution of endogenous VDPs via ratiometric fluorescence imaging and colocalization assay. And the results suggested that there are abundant VDPs in mitochondria. Moreover, some redox-sensitive VDPs are also present on cell surface which can respond to redox stimulus. This ratiometric fluorescence technique presents an important extension to previous fluorescence intensity-based probes for trapping and quantifying protein vicinal dithiols in living cells, as well as its visible dynamic tracing of VDPs.