Kawagoe Ryosuke, Takashima Ippei, Uchinomiya Shohei, Ojida Akio
Graduate School of Pharmaceutical Sciences , Kyushu University , 3-1-1 Maidashi, Higashi-ku , Fukuoka , 812-8582 , Japan.
Chem Sci. 2017 Feb 1;8(2):1134-1140. doi: 10.1039/c6sc03856e. Epub 2016 Sep 26.
Hydropersulfide (R-SSH) is an important class of reactive sulfur species (RSS) involved in a variety of physiological processes in mammals. A fluorescent probe capable of real-time detection of hydropersulfide levels in living cells would be a versatile tool to elucidate its roles in cell signalling and redox homeostasis. In this paper, we report a ratiometric fluorescent probe for hydropersulfide sensing, based on a fluorescence resonance energy transfer (FRET) mechanism. This sensing mechanism involves a nucleophilic reaction of a hydropersulfide with the pyronine-unit of the probe, which modulates the intramolecular FRET efficiency to induce a dual-emission change. The reversible nature of this reaction allows us to detect increases and decreases of hydropersulfide levels in a real-time manner. The probe fluorometrically sensed highly reactive hydropersulfides, such as HS and Cys-SSH, while the fluorescence response to biologically abundant cysteine and glutathione was negligible. Taking advantage of the reversible and selective sensing properties, this probe was successfully applied to the ratiometric imaging of concentration dynamics of endogenously produced hydropersulfides in living cells.
氢过硫化物（R-SSH）是一类重要的活性硫物质（RSS），参与哺乳动物的多种生理过程。一种能够实时检测活细胞中氢过硫化物水平的荧光探针将是阐明其在细胞信号传导和氧化还原稳态中作用的多功能工具。在本文中，我们报道了一种基于荧光共振能量转移（FRET）机制的用于氢过硫化物传感的比率荧光探针。这种传感机制涉及氢过硫化物与探针的吡咯宁单元的亲核反应，该反应调节分子内FRET效率以诱导双发射变化。该反应的可逆性使我们能够实时检测氢过硫化物水平的升高和降低。该探针通过荧光法检测高活性氢过硫化物，如HS和Cys-SSH，而对生物体内丰富的半胱氨酸和谷胱甘肽的荧光响应可忽略不计。利用这种可逆和选择性的传感特性，该探针成功应用于活细胞内源性产生的氢过硫化物浓度动态的比率成像。
