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Development of Acrylamide-Based Rapid and Multicolour Fluorogenic Probes for High Signal-to-Noise Living Cell Imaging.

Research paper by Dasheng D Zhang, Renmei R Liu, Chunyan C Bao, Chenxia C Zhang, Lipeng L Yang, Lei L Deng, Bingkun B Bao, Jing J Yang, Xianjun X Chen, Qiuning Q Lin, Yi Y Yang, Linyong L Zhu

Indexed on: 20 Dec '18Published on: 20 Dec '18Published in: Bioconjugate Chemistry



Abstract

Protein covalent labelling is dramatically useful for studying protein function in living cells and organisms. In this field, chemical tag technique combines with fluorogenic probes have emerged as a powerful tool. Herein a series of TMP-tag fluorogenic probes have been developed for spanning green to full blue spectral range. These probes feature an acrylamide unite which 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 unite specifically react with the thiol group of L28C residue beside the ligand binding pocket, achieving protein-specific labelling without any liberation of leaving groups. With these probes, multicolour and specific protein labelling with fast reaction rate (t1/2=33 s) and dramatic fluorescence enhancement (4000-fold) was obtained. Furthermore, no-wash protein labelling of both in living cells and in 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.