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Activatable Formation of Emissive Excimers for Highly Selective Detection of β-Galactosidase.
Analytical Chemistry ( IF 6.7 ) Pub Date : 2020-03-31 , DOI: 10.1021/acs.analchem.9b04806
Yang Li 1 , Lulu Ning 2 , Fang Yuan 1 , Tian Zhang 3 , Jianjian Zhang 1 , Zhigang Xu 3 , Xiao-Feng Yang 1
Affiliation  

Small-molecular fluorescence sensors have become promising detection tools in many fields attributing to their high sensitivity, excellent temporal and spatial resolution, and low cytotoxicity. However, high concentration or aggregation-induced fluorescence quenching effect has usually hindered the development of traditional fluorescence dyes. Herein, a new fluorophore cyanovinylene dye BMZ with excimer emission property has been constructed. It shows an obvious enhanced and red-shift emission upon aggregation in aqueous solution, which overmatches the conventional pyrene with longer absorption and emission wavelengths. Using this unique optical property, a new fluorescence probe BMZ-Gal has been developed for trapping of β-galactosidase (β-Gal) activity with high selectivity, low limit of detection of 0.17 U, and rapid recognition, which is based on the β-Gal-induced formation of red-shift emitting excimer. β-Gal has a strong affinity for BMZ-Gal, which is verified through the Michaelis-Menten constants (Km, 1.87 μM) and the hydrolysis efficiencies (Kcat/Km, 1.78 × 103 M-1 s-1). Furthermore, the assay system has been successfully used for detecting endogenous β-Gal in living ovarian cancer cells and can passively targeted to identify β-Gal in organelle level and determine its subcellular location with satisfactory accuracy. We anticipate that the new fluorophore cyanovinylene dye herein may inaugurate new opportunities for the development of excimer emission sensors.

中文翻译:

可选择性地检测β-半乳糖苷酶的发射性准分子的可活化形成。

小分子荧光传感器因其高灵敏度,出色的时间和空间分辨率以及低细胞毒性而在许多领域中成为有前途的检测工具。然而,高浓度或聚集诱导的荧光猝灭作用通常阻碍了传统荧光染料的发展。在此,已经构造了具有准分子发射性能的新型荧光团氰基亚乙烯基染料BMZ。它在水溶液中聚集后显示出明显的增强和红移发射,这与具有更长吸收和发射波长的常规pyr不匹配。利用这种独特的光学特性,开发了一种新型荧光探针BMZ-Gal,用于捕获β-半乳糖苷酶(β-Gal)活性,具有高选择性,0.17 U的低检出限和快速识别,这是基于β-Gal诱导的发红移准分子的形成。β-Gal对BMZ-Gal具有很强的亲和力,这已通过Michaelis-Menten常数(Km,1.87μM)和水解效率(Kcat / Km,1.78×103 M-1 s-1)进行了验证。此外,该测定系统已经成功地用于检测活的卵巢癌细胞中的内源性β-Gal,并且可以被动地靶向鉴定细胞器水平中的β-Gal,并以令人满意的精度确定其亚细胞位置。我们预期本文中的新型荧光团氰基亚乙烯基染料可能会为准分子发射传感器的开发提供新的机会。87μM)和水解效率(Kcat / Km,1.78×103 M-1 s-1)。此外,该测定系统已经成功地用于检测活的卵巢癌细胞中的内源性β-Gal,并且可以被动地靶向鉴定细胞器水平中的β-Gal,并以令人满意的精度确定其亚细胞位置。我们预期本文中的新的荧光团氰基亚乙烯基染料可能会为准分子发射传感器的开发提供新的机会。87μM)和水解效率(Kcat / Km,1.78×103 M-1 s-1)。此外,该测定系统已经成功地用于检测活的卵巢癌细胞中的内源性β-Gal,并且可以被动地靶向鉴定细胞器水平中的β-Gal,并以令人满意的精度确定其亚细胞位置。我们预期本文中的新型荧光团氰基亚乙烯基染料可能会为准分子发射传感器的开发提供新的机会。
更新日期:2020-04-23
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