A coumarin-based fluorescence sensor for rapid discrimination of cysteine/homocysteine and glutathione under dual excitation wavelengths

Abstract

Biological thiols (Cys, Hcy and GSH) are crucial biomolecules in living cells and play indispensable roles in maintaining the redox homeostasis of organisms. But due to their similar molecular structure, the development of effective tools for distinguishing two or three of them remains a great difficulty. Herein, we constructed a sensitive sensor (CB) by connecting the bifunctional fluorescent reagent with coumarin derivatives for simultaneous recognition of these three thiols through different pathways. Free CB had no fluorescence; however, with gradual addition of thiols, the chlorine unit was replaced by sulfhydryl. Furthermore, the intramolecular rearrangement occurred between the amino and sulfhydryl groups of Cys/Hcy and yellow fluorescence was observed at 570 nm. However, GSH with a large structure could not undergo intramolecular rearrangement, and green fluorescence was excited at 505 nm. In this way, Cys/Hcy and GSH can be detected distinctively. Under dual excitation wavelengths, CB exhibited high selectivity and fast response to the three thiols. Furthermore, CB was successfully applied to imaging endogenous and exogenous thiols in living cells and zebrafish, providing us with a reliable tool for thiols recognition.