Molecular Rotor-Based Fluorescent Probe for Selective Recognition of Hybrid G-Quadruplex and as a K+ Sensor

化学 荧光 G-四倍体 堆积 苯并噻唑 硫黄素 DNA 结晶学 生物化学 量子力学 医学 物理 病理 有机化学 阿尔茨海默病 疾病
作者
Lingling Liu,Yong Shao,Jian Peng,Chaobiao Huang,Hua Liu,Lihua Zhang
出处
期刊:Analytical Chemistry [American Chemical Society]
卷期号:86 (3): 1622-1631 被引量:113
标识
DOI:10.1021/ac403326m
摘要

This work demonstrates the significant fluorescence enhancement of thioflavin T (ThT) when binding to G-quadruplexes possessing hybrid structures by using UV–vis absorption spectra, fluorescence spectra, and Tm experiments to confirm the binding events. ThT binding does not disturb native G-quadruplex structures preformed in Na+ and K+ solutions. The fluorescence enhancement is caused by the rotation restriction of benzothiazole (BZT) and dimethylaminobenzene (DMAB) rings in the ThT excited state upon its G-quadruplex binding. This molecular rotor mechanism as a means of fluorescence enhancement is confirmed using a nonrotor analogue of ThT. Hydroxylation and electrolyte experiments demonstrate that ThT stacks on the tetrad of the hybrid G-quadruplexes, whereas electrostatic forces contribute more to ThT binding for other G-quadruplex structures. By stacking on the tetrad, the ThT binding favors selective identification of DNA hybrid G-quadruplex structures with enhanced fluorescence and can serve as a conformation probe to monitor G-quadruplex structure conversion between hybrid and other structures. Using these properties, we developed a selective and label-free fluorescent K+ sensor with a detection limit of 1 mM for K+ in the presence of 100 mM Na+. The coexistence of other metal ions produces a fluorescence response comparable to K+ alone. We believe that ThT can potentially provide structure identification of hybrid G-quadruplexes and aid in the construction of G-quadruplex-based sensors.
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