Department of Applied Chemistry, National Chiao Tung University, Hsinchu 300, Taiwan.
Department of Applied Chemistry, National Chiao Tung University, Hsinchu 300, Taiwan.
Spectrochim Acta A Mol Biomol Spectrosc. 2017 Aug 5;183:204-208. doi: 10.1016/j.saa.2017.04.051. Epub 2017 Apr 20.
Cysteine (Cys) and homocysteine (Hcy) are two of important biological thiols and function as important roles in several biological processes. The development of Cys and Hcy probes will help to explore the functions of biothiols in biological systems. In this work, a new coumarin-based probe AC, containing an acryloyl moiety, was developed for Cys and Hcy detection in cells. Cys and Hcy undergo a nucleophilic addition and subsequent cyclization reaction to remove to the acryloyl group and yield a fluorescent product, 7-hydroxylcomuarin. The probe AC showed good selectivity for cysteine and homocysteine over glutathione and other amino acids and had low detection limits of 65nM for Cys and 79nM for Hcy, respectively. Additionally, confocal imaging experiments demonstrated that the probe AC can be applied to visualize Cys and Hcy in living cells.
半胱氨酸（Cys）和同型半胱氨酸（Hcy）是两种重要的生物硫醇，在几种生物过程中发挥着重要作用。Cys 和 Hcy 探针的发展将有助于探索生物体系中生物硫醇的功能。在这项工作中，开发了一种基于香豆素的新型探针 AC，其含有丙烯酰基部分，可用于细胞中 Cys 和 Hcy 的检测。Cys 和 Hcy 通过亲核加成和随后的环化反应去除丙烯酰基，生成荧光产物 7-羟基香豆素。探针 AC 对半胱氨酸和同型半胱氨酸具有良好的选择性，对谷胱甘肽和其他氨基酸的选择性较低，对 Cys 的检测限分别为 65nM，对 Hcy 的检测限分别为 79nM。此外，共聚焦成像实验表明，探针 AC 可用于可视化活细胞中的 Cys 和 Hcy。
