Key Laboratory for Advanced Materials and Institute of Fine Chemicals, School of Chemistry and Molecular Engineering, East China University of Science & Technology, Shanghai 200237, PR China.
Key Laboratory for Advanced Materials and Institute of Fine Chemicals, School of Chemistry and Molecular Engineering, East China University of Science & Technology, Shanghai 200237, PR China; National Center for Drug Screening, State Key Laboratory of Drug Research, Shanghai, Institute of Materia Medica, Chinese Academy of Sciences, 189 Guo Shoujing Rd., Shanghai 201203, PR China.
Spectrochim Acta A Mol Biomol Spectrosc. 2021 Nov 15;261:120055. doi: 10.1016/j.saa.2021.120055. Epub 2021 Jun 5.
Biothiols including cysteine (Cys) and glutathione (GSH) are biological signaling molecules responsible for cell detoxification, cell metabolism and neutralization of reactive oxygen species. Here, we synthesized a long-wavelength fluorescent probe, DCIMA, for lysosome-targeted imaging of Cys and GSH in living cells. DCIMA is consisted of a dicyanoisophorone core modified with an acrylate group for biothiol detection through the Michael addition reaction and a morpholine group as the lysosome-targeting agent. The presence of the electron-donating morpholine group also enhances the intramolecular charge transfer mechanism of the probe, thereby enabling its long-wavelength fluorescence emission (670 nm) and large Stokes shift (180 nm). In concentration range of 0-30 μM, the probe was determined to react quickly with both Cys and GSH with low detection limits (<5 min, 35.2 nM for GSH and 34.8 nM for Cys) and achieve the sensitive fluorescence imaging of the biothiols located in the lysosomes of living cells.
生物硫醇包括半胱氨酸（Cys）和谷胱甘肽（GSH），是负责细胞解毒、细胞代谢和中和活性氧的生物信号分子。在这里，我们合成了一种长波长荧光探针 DCIMA，用于活细胞中溶酶体靶向的 Cys 和 GSH 成像。DCIMA 由一个二氰基异佛尔酮核心组成，该核心通过迈克尔加成反应修饰有一个丙烯酰基，用于通过生物硫醇检测，以及一个吗啉基团作为溶酶体靶向剂。供电子吗啉基团的存在也增强了探针的分子内电荷转移机制，从而使探针能够发射长波长荧光（670nm）和大斯托克斯位移（180nm）。在 0-30μM 的浓度范围内，该探针被确定能够快速与 Cys 和 GSH 反应，检测限低（<5 分钟，GSH 的检测限为 35.2nM，Cys 的检测限为 34.8nM），并实现了位于活细胞溶酶体中的生物硫醇的灵敏荧光成像。
