Department of Chemistry, The University of Texas at Austin , 105 East 24th Street, Stop A5300, Austin, Texas 78712-1224, United States.
J Am Chem Soc. 2018 Feb 7;140(5):1870-1875. doi: 10.1021/jacs.7b12545. Epub 2018 Jan 25.
An ability to visualize HCN in mitochondria in real time may permit additional insights into the critical toxicological and physiological roles this classic toxin plays in living organisms. Herein, we report a mitochondria-specific coumarin pyrrolidinium-derived fluorescence probe (MRP1) that permits the real-time ratiometric imaging of HCN in living cells. The response is specific, sensitive (detection limit is ca. 65.6 nM), rapid (within 1 s), and reversible. Probe MRP1 contains a benzyl chloride subunit designed to enhance retention within the mitochondria under conditions where the mitochondria membrane potential is eliminated. It has proved effective in visualizing different concentrations of exogenous HCN in the mitochondria of HepG2 cells, as well as the imaging of endogenous HCN in the mitochondria of PC12 cells and within neurons. Fluctuations in HCN levels arising from the intracellular generation of HCN could be readily detected.
实时可视化 HCN 在线粒体中的能力可能会使我们对这种经典毒素在生物体内的关键毒理学和生理学作用有更多的了解。在此，我们报告了一种线粒体特异性香豆素吡咯烷鎓衍生的荧光探针（MRP1），该探针允许在活细胞中实时比率成像 HCN。该响应具有特异性、灵敏度高（检测限约为 65.6 nM）、快速（在 1 秒内）和可逆性。探针 MRP1 包含一个苄基氯亚基，旨在消除线粒体膜电位时增强其在线粒体中的保留。它已被证明可有效可视化 HepG2 细胞线粒体中不同浓度的外源 HCN，以及 PC12 细胞线粒体和神经元内内源 HCN 的成像。很容易检测到由细胞内 HCN 生成引起的 HCN 水平波动。
