Li Xiaolu, Feng Qian, Qu Lejing, Zhao Ting, Li Xiaoan, Bai Tiantian, Sun Shisheng, Wu Shaoping, Zhang Yongmin, Li Jianli
Key Laboratory of Resource Biology and Biotechnology in Western China (Northwest University), Ministry of Education, Biomedicine Key Laboratory of Shaanxi Province, Northwest University, Xi'an, 710069, PR China; Joint International Laboratory of Glycobiology and Medicinal Chemistry, Northwest University, Xi'an, Shaanxi, 710069, PR China.
Key Laboratory of Resource Biology and Biotechnology in Western China (Northwest University), Ministry of Education, Biomedicine Key Laboratory of Shaanxi Province, Northwest University, Xi'an, 710069, PR China.
Anal Chim Acta. 2020 Aug 22;1126:72-81. doi: 10.1016/j.aca.2020.06.026. Epub 2020 Jun 18.
The amount of protein thiols play a crucial role in maintaining the cellular redox homeostasis and have significant implications to indicate a series of diseases. Therefore, it is necessary to develop an ideal probe for protein thiol detection in a simple and readily implementable method. Consequently, a water-soluble and incubate-free fluorescent environment-sensitive probe DMTs-OCC was synthesized using 7-diethylamincoumarin as the fluorophore and 4-(5-Methanesulfonyl- [1,2,3,4]tetrazol-1-yl)-phenol (MSTP) as a thiol receptor reagent. The blue-shift emission spectra of probe DMTs-OCC was observed by ultrafast binding to protein sulfhydryl groups from the excited intramolecular charge transfer (ICT) to the twisted intramolecular charge transfer (TICT) conversion process in aqueous solution. The experimental results showed that probe DMTs-OCC exhibited an excellent selectivity to protein thiols and biocompatibility in aqueous solution, as well as terrific cell membrane permeability which enabled the successful visualization of BSA protein thiol in living cells. Moreover, no excess probe was cleaned and no incubation time was needed in cell experiments. Therefore, it could provide a new method to the construction of fluorescent probes for protein thiols labelling and visualization.
蛋白质硫醇的含量在维持细胞氧化还原稳态中起着关键作用,对一系列疾病的指示具有重要意义。因此,有必要开发一种以简单且易于实施的方法检测蛋白质硫醇的理想探针。为此,以7-二乙氨基香豆素作为荧光团,4-(5-甲磺酰基-[1,2,3,4]四唑-1-基)苯酚(MSTP)作为硫醇受体试剂,合成了一种水溶性且无需孵育的荧光环境敏感探针DMTs-OCC。在水溶液中,通过从激发态分子内电荷转移(ICT)到扭曲分子内电荷转移(TICT)的转换过程与蛋白质巯基超快结合,观察到探针DMTs-OCC的蓝移发射光谱。实验结果表明,探针DMTs-OCC在水溶液中对蛋白质硫醇表现出优异的选择性和生物相容性,以及出色的细胞膜通透性,能够成功实现活细胞中牛血清白蛋白(BSA)蛋白质硫醇的可视化。此外,细胞实验中无需清洗多余的探针,也无需孵育时间。因此,它可为构建用于蛋白质硫醇标记和可视化的荧光探针提供一种新方法。