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一种用于高选择性和灵敏检测活细胞中含邻二硫醇蛋白质的红色荧光点亮探针的研发。

Development of a red fluorescent light-up probe for highly selective and sensitive detection of vicinal dithiol-containing proteins in living cells.

作者信息

Wang Yuanyuan, Yang Xiao-Feng, Zhong Yaogang, Gong Xueyun, Li Zheng, Li Hua

机构信息

Key Laboratory of Synthetic and Natural Functional Molecule Chemistry of Ministry of Education , College of Chemistry & Materials Science , Northwest University , Xi'an 710069 , P. R. China . Email:

College of Life Sciences , Northwest University , Xi'an 710069 , P. R. China.

出版信息

Chem Sci. 2016 Jan 1;7(1):518-524. doi: 10.1039/c5sc02824h. Epub 2015 Oct 23.

Abstract

Vicinal dithiol-containing proteins (VDPs) play a key role in cellular redox homeostasis and are responsible for many diseases. Here, we develop a red fluorescent light-up probe for the highly selective and sensitive detection of VDPs using the environment-sensitive 2-(4-dimethylaminophenyl)-4-(2-carboxyphenyl)-7-diethylamino-1-benzopyrylium () as the fluorescent reporter and cyclic dithiaarsane as the targeting unit. is almost nonfluorescent in aqueous solution. However, it exhibits intense fluorescence emission upon binding to reduced bovine serum albumin (rBSA, selected as the model protein). The fluorescence intensity of is directly proportional to the concentration of rBSA over the range of 0.06-0.9 μM, with a detection limit (3) of 0.015 μM. Importantly, the fast kinetics of binding between and VDPs (∼2.5 min) enables the dynamic tracing of VDPs in biological systems. Preliminary experiments show that can be used for the no-wash imaging of endogenous VDPs in living cells. In addition, our study shows that presents both high environment-sensitivity and good fluorescence properties, and is promising for the development of no-wash fluorescent light-up probes for target-specific proteins in living cells.

摘要

含邻位二硫醇的蛋白质(VDPs)在细胞氧化还原稳态中起关键作用,并与多种疾病相关。在此,我们开发了一种红色荧光点亮探针,用于高选择性和灵敏地检测VDPs。该探针使用对环境敏感的2-(4-二甲基氨基苯基)-4-(2-羧基苯基)-7-二乙氨基-1-苯并吡喃鎓()作为荧光报告基团,环二硫砷烷作为靶向单元。在水溶液中几乎无荧光。然而,它与还原型牛血清白蛋白(rBSA,选为模型蛋白)结合后会发出强烈的荧光。在0.06 - 0.9 μM范围内,的荧光强度与rBSA的浓度成正比,检测限(3)为0.015 μM。重要的是,与VDPs之间快速的结合动力学(约2.5分钟)能够动态追踪生物系统中的VDPs。初步实验表明,可用于活细胞内源性VDPs的免洗成像。此外,我们的研究表明,具有高环境敏感性和良好的荧光特性,有望用于开发活细胞中靶标特异性蛋白质的免洗荧光点亮探针。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b30d/5519953/86e697a7b051/c5sc02824h-s1.jpg

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