Ye Haishun, Cheng Longhuai, Tu Xiaoqiang, Wang Da-Wei, Yi Long
State Key Laboratory of Organic-Inorganic Composites and Beijing Key Lab of Bioprocess, Beijing University of Chemical Technology (BUCT) Beijing 100029 China
State Key Laboratory of Elemento-Organic Chemistry and Department of Chemical Biology, College of Chemistry, National Pesticide Engineering Research Center, Nankai University Tianjin 300071 China
RSC Chem Biol. 2022 May 17;3(7):848-852. doi: 10.1039/d2cb00105e. eCollection 2022 Jul 6.
Biothiols and their interconversion are involved in cellular redox homeostasis as well as many physiological processes. Here, a dual-reactive dual-quenching fluorescent probe was rationally developed based on thiolysis reactions of 7-nitrobenzoxadiazole (NBD) tertiary amine and 7-cyanobenzoxadiazole (CBD) arylether for imaging of the biothiol interconversion. We demonstrate that the NBD-CBD probe exhibits very weak background fluorescence due to the dual-quenching effects, and can be dual-activated by HS and GSH with an over 500-fold fluorescence increase at 525 nm. In addition, the probe shows high sensitivity, excellent selectivity, and good biocompatibility, all of which promote the simultaneous detection of both HS and GSH in live cells. Importantly, probe 1 was successfully employed to reveal the biogenesis of both HS and GSH from l-Cys rather than from d-Cys, and therefore, d-Cys would be solely converted into HS, which may help understand the more HS generation from d-Cys than from l-Cys in live cells.
生物硫醇及其相互转化参与细胞氧化还原稳态以及许多生理过程。在此，基于7-硝基苯并恶二唑（NBD）叔胺和7-氰基苯并恶二唑（CBD）芳基醚的硫解反应，合理开发了一种双反应双猝灭荧光探针，用于生物硫醇相互转化的成像。我们证明，由于双猝灭效应，NBD-CBD探针表现出非常弱的背景荧光，并且可以被HS和GSH双重激活，在525 nm处荧光增强超过500倍。此外，该探针具有高灵敏度、优异的选择性和良好的生物相容性，所有这些都有助于在活细胞中同时检测HS和GSH。重要的是，探针1成功用于揭示l-半胱氨酸而非d-半胱氨酸产生HS和GSH的生物合成过程，因此，d-半胱氨酸将仅转化为HS，这可能有助于理解活细胞中d-半胱氨酸比l-半胱氨酸产生更多HS 的原因。
