使用基于荧光亲和力的探针区分链霉亲和素和抗生物素蛋白。

Discrimination between streptavidin and avidin with fluorescent affinity-based probes.

作者信息

Sun Qian, Tian Haiyu, Qu Haoran, Sun Deheng, Chen Zhuo, Duan Liping, Zhang Weibing, Qian Junhong

机构信息

Shanghai Key Laboratory of Functional Materials Chemistry, School of Chemistry and Molecular Engineering, East China University of Science and Technology, Shanghai, 200237, China.

出版信息

Analyst. 2015 Jul 7;140(13):4648-53. doi: 10.1039/c5an00585j.

DOI:10.1039/c5an00585j

PMID:25985268

Abstract

Two biotinylated coumarin-based fluorescent probes SPS3 and RC3 were designed for differentiating between structurally similar proteins streptavidin (SA) and avidin (AV). A substituted phenyl group is introduced onto SPS3, which may quench the fluorescence through twist intramolecular charge transfer (TICT). The fluorescence of SPS3 is turned on, by restraining the TICT process, when the fluorophore is buried at the surface of SA. RC3 is constructed by incorporating a biotin molecule to a coumarin fluorophore through a 4-atom spacer. The fluorescence intensity of RC3 is enhanced significantly when its fluorophore enters into the less polar binding pocket of AV. SPS3 and RC3 could be applied in distinguishing between SA and AV as well as in fluorescence imaging of biotin receptor over-expressed Hela cells.

摘要

设计了两种基于香豆素的生物素化荧光探针SPS3和RC3，用于区分结构相似的蛋白质链霉亲和素（SA）和抗生物素蛋白（AV）。在SPS3上引入了一个取代苯基，其可能通过扭曲分子内电荷转移（TICT）淬灭荧光。当荧光团埋在SA表面时，通过抑制TICT过程，SPS3的荧光被开启。RC3是通过一个4原子间隔基将生物素分子连接到香豆素荧光团上构建而成。当RC3的荧光团进入AV的极性较小的结合口袋时，其荧光强度显著增强。SPS3和RC3可用于区分SA和AV以及对生物素受体过表达的Hela细胞进行荧光成像。