Department of Biophysical Chemistry, J. Heyrovský Institute of Physical Chemistry of the CAS, v. v. i., Dolejškova 2155/3, 182 23 Prague 8, Czechia.
Methods Appl Fluoresc. 2018 Feb 12;6(2):025005. doi: 10.1088/2050-6120/aaaa05.
Coumarin derivatives are well known fluorescence reporters for investigating biological systems due to their strong micro-environment sensitivity. Despite having wide range of environment sensitive fluorescence probes, the potential of 6,7-dimethoxy-coumarin has not been studied extensively so far. With a perspective of its use in protein studies, namely using the unnatural amino acid technology or as a substrate for hydrolase enzymes, we study acetyloxymethyl-6,7-dimethoxycoumarin (Ac-DMC). We investigate the photophysics and hydration dynamics of this dye in aerosol-OT (AOT) reverse micelles at various water contents using the time dependent fluorescence shift (TDFS) method. The TDFS response in AOT reverse micelles from water/surfactant ratio of 0 to 20 confirms its sensitivity towards the hydration and mobility of its microenvironment. Moreover, we show that the fluorophore can be efficiently quenched by halide ions. Hence, we conclude that the 6,7-dimethoxy-methylcoumarin fluorophore is useful for studying hydration parameters in biologically relevant systems.
香豆素衍生物由于其对微环境的高灵敏度而被广泛用作研究生物体系的荧光报告基团。尽管已经有了广泛的环境敏感荧光探针,但 6,7-二甲氧基香豆素的潜力迄今为止尚未得到广泛研究。从其在蛋白质研究中的应用角度来看,即使用非天然氨基酸技术或作为水解酶的底物,我们研究了乙酰氧甲基-6,7-二甲氧基香豆素(Ac-DMC)。我们使用时间分辨荧光位移(TDFS)方法研究了该染料在气溶胶 OT(AOT)反向胶束中不同含水量下的光物理和水合动力学。从水/表面活性剂比为 0 到 20 的 AOT 反向胶束中的 TDFS 响应证实了其对微环境水合作用和迁移率的敏感性。此外,我们还表明,卤化物离子可以有效地猝灭荧光团。因此,我们得出结论,6,7-二甲氧基甲基香豆素荧光团可用于研究生物相关体系中的水合参数。