文献类型：期刊文献

英文题名：Development of Acrylamide-Based Rapid and Multicolor Fluorogenic Probes for High Signal-to-Noise Live Cell Imaging

作者：Zhang, Dasheng[1,2];Liu, Renmei[1,3];Bao, Chunyan[1,2];Zhang, Chenxia[1,3];Yang, Lipeng[1,2];Deng, Lei[1,3];Bao, Bingkun[1,2];Yang, Jing[1,3];Chen, Xianjun[1,3];Lin, Qiuning[1,2];Yang, Yi[1,3];Zhu, Linyong[1,2]

机构：[1]East China Univ Sci & Technol, Shanghai Collaborat Innovat Ctr Biomfg Technol, State Key Lab Bioreactor Engn, Optogenet & Synthet Biol Interdisciplinary Res Ct, 130 Mei Long Rd, Shanghai 200237, Peoples R China;[2]East China Univ Sci & Technol, Sch Chem & Mol Engn, 130 Mei Long Rd, Shanghai 200237, Peoples R China;[3]East China Univ Sci & Technol, Sch Pharm, 130 Mei Long Rd, Shanghai 200237, Peoples R China

年份：2019

卷号：30

期号：1

起止页码：184

外文期刊名：BIOCONJUGATE CHEMISTRY

收录：;EI(收录号：20220411530792);WOS:【SCI-EXPANDED(收录号:WOS:000456350800019)】；

基金：This work was supported by the NSFC (Grants 21425311, 21472044, 31225008, 31470833, and 51403061), the Fundamental Research Funds for the Central Universities (222201717003), the National Key Research and Development Program of China (2017YFA050400), and The State Key Laboratory of Bioreactor Engineering.

语种：英文

外文关键词：Amides - Cells - Proteins - Signal to noise ratio - Ligands - Cytology

摘要：Protein covalent labeling is dramatically useful for studying protein function in living cells and organisms. In this field, the chemical tag technique combined with fluorogenic probes has emerged as a powerful tool. Herein, a series of TMP tag fluorogenic probes have been developed to span the green to full blue spectral range. These probes feature an acrylamide unit that acts as a linker group to conjugate the fluorophore and the ligand as well as a quencher and a covalent reaction group. After the probes bind to eDHFR:L28C, the acrylamide unit specifically reacts with the thiol group of the L28C residue beside the ligand binding pocket, achieving protein-specific labeling without any liberation of leaving groups. With these probes, multicolor and specific protein labeling with a fast reaction rate (t(1/2) = 33 s) and dramatic fluorescence enhancement (4000-fold) were obtained. Furthermore, no-wash protein labeling in both living cells and zebrafish was successfully achieved. We expect it may provide a general and highly effective chemical tool for the study of protein function in living cells and organisms.