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