State Key Laboratory of Organic-Inorganic Composites and Beijing Key Lab of Bioprocess, Beijing University of Chemical Technology, Beijing 100029, China.
Tianjin Key Laboratory of Technologies Enabling Development of Clinical Therapeutics and Diagnostics (Theranostics), School of Pharmacy, Tianjin Medical University, Tianjin 300070, China.
Anal Chem. 2022 Jan 25;94(3):1733-1741. doi: 10.1021/acs.analchem.1c04324. Epub 2022 Jan 12.
HS is a gaseous signaling molecule that is involved in many physiological and pathological processes. In general, the level of intracellular HS (<1 μM) is much lower than that of GSH (∼1-10 mM), leading to the remaining challenge of selective detection and differentiation of endogenous HS in live biosystems. To this end, we quantitatively demonstrate that the thiolysis of NBD amine has much higher selectivity for HS over GSH than that of the reduction of aryl azide. Subsequently, we developed the first NBD-based cell-trappable probe () for highly selective and ultrasensitive imaging of intracellular HS. Probe demonstrates a 207-fold fluorescence enhancement at 520 nm after reaction with HS/esterase to produce a bright BODIPY (quantum yield 0.42) and a detection limit of 15.7 nM. Probe is water-soluble, cell-trappable, and not cytotoxic. Based on this excellent chemical tool, relative levels of basal HS in different cell lines were measured to reveal a positive correlation between endogenous HS and the metastatic potential of colon and breast cancer cells. In addition, HS biogenesis was also validated by probe both in tobacco leaves under viral infection and in zebrafish after tail amputation. It is anticipated that probe will have widespread applications in imaging and for investigating different HS-related biological processes and diseases.
HS 是一种气态信号分子，参与许多生理和病理过程。一般来说，细胞内 HS 的水平（<1 μM）远低于 GSH（∼1-10 mM），这导致在活生物体系中选择性检测和区分内源性 HS 仍然具有挑战性。为此，我们定量证明了 NBD 胺的硫解对 HS 的选择性比芳基叠氮化物的还原高得多。随后，我们开发了第一个基于 NBD 的可细胞捕获探针 ()，用于高度选择性和超灵敏地检测细胞内 HS。探针在与 HS/酯酶反应后，在 520nm 处产生明亮的 BODIPY（量子产率为 0.42），荧光增强 207 倍，检测限为 15.7 nM。探针水溶性好，可细胞捕获，且无细胞毒性。基于这个出色的化学工具，我们测量了不同细胞系中的基础 HS 水平，以揭示内源性 HS 与结肠癌和乳腺癌细胞转移潜能之间的正相关关系。此外，探针还分别在烟草叶片受到病毒感染和斑马鱼尾部截断后验证了 HS 的生物合成。预计探针将广泛应用于成像以及研究不同的 HS 相关生物过程和疾病。
