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

• Published in: Bioconjugate chemistry 2019-01, Vol.30 (1), p.184-191
• Main Authors: Zhang, Dasheng, Liu, Renmei, Bao, Chunyan, Zhang, Chenxia, Yang, Lipeng, Deng, Lei, Bao, Bingkun, Yang, Jing, Chen, Xianjun, Lin, Qiuning, Yang, Yi, Zhu, Linyong
• Format: Article
• Language: English
• Subjects: Acrylamide; Acrylamide - chemistry; Acrylamide - metabolism; Animals; Cell Nucleus - metabolism; Cells; Cells (biology); Fluorescence; Fluorescent Dyes - chemistry; Fluorescent Dyes - metabolism; HEK293 Cells; HeLa Cells; Humans; Labeling; Ligands; Molecular Imaging - methods; Organic chemistry; Probes; Protein folding; Proteins; Signal-To-Noise Ratio; Tetrahydrofolate Dehydrogenase - genetics; Zebrafish
• ISSN: 1043-1802; 1520-4812
• DOI: 10.1021/acs.bioconjchem.8b00827

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.