A thiol-anchored solvatochromic and fluorogenic molecular rotor for covalent protein labeling in SDS-PAGE and mitochondria specific fluorescence imaging.

Fluorescent labeling is a widely used method for protein detection and fluorescence imaging. A solvatochromic and fluorogenic molecular rotor DASPBCl was developed for covalent protein labeling in solution and SDS-PAGE, and also for stable mitochondria labeling and fluorescence imaging. The dye DASPBCl consisted of a 4-(N,N-dimethylamino)phenyl moiety as the electron donor and a positively charged N-benzylpyridinium moiety as the electron acceptor. A benzyl chloride group was introduced into the pyridine moiety for covalent labeling of thiol in proteins. When the fluorescent dye DASPBCl is covalently labeled to the thiol of proteins, significantly enhanced fluorescence was obtained, which is attributed to the polarity sensitivity caused solvatochromic effect from the hydrophobic protein structure and the viscosity sensitivity caused fluorogenic effect from the restriction of single bond rotation. DASPBCl exhibits high sensitivity and good linear response for protein detection in SDS-PAGE analysis with both the pre-staining method and post-staining method. DASPBCl was also successfully used for covalently protein-anchored fluorescence imaging of mitochondria in living cells.

An aggregation induced emission chalcone fluorescent probe with large Stokes shift for biothiols detection.

The homeostasis of biothiols is closely related to the health of organisms. In view of the important role of biothiols, a fluorescent probe (7HIN-D) for the detection of intracellular biothiols was developed based on a simple chalcone fluorophore 7HIN with "ESIPT + AIE" characteristics. The probe 7HIN-D was obtained by introducing a biothiols specific DNBS (2,4-dinitrobenzenesulfonyl) unit as a fluorescence quencher to the fluorophore 7HIN. The nucleophilic substitution reaction between biothiols and probe 7HIN-D will release the DNBS unit and the fluorophore 7HIN, which exhibits a "turn on" AIE fluorescence with a large Stokes shift of 113 nm. The probe 7HIN-D displays high sensitivity and good selectivity to biothiols, and the detection limits value of probe 7HIN-D for GSH, Cys and Hcy were 0.384 µmol/L, 0.471 µmol/L and 0.638 µmol/L, respectively. In addition, the probe has been successfully used for fluorescence detection of endogenous biothiols in living cells due to its excellent performance, good biocompatibility and low cytotoxicity.