通过控制荧光闪烁对核酸结构切换动力学的单分子监测。
Single-Molecule Monitoring of the Structural Switching Dynamics of Nucleic Acids through Controlling Fluorescence Blinking.
机构信息
The Institute of Scientific and Industrial Research (SANKEN), Osaka University, Mihogaoka 8-1, Ibaraki, Osaka, 567-0047, Japan.
Department of Life Science and Technology, Tokyo Institute of Technology, 4259 B-57 Nagatsuta, Midori-ku, Yokohama, Kanagawa, 226-8501, Japan.
出版信息
Angew Chem Int Ed Engl. 2017 Nov 27;56(48):15329-15333. doi: 10.1002/anie.201708705. Epub 2017 Oct 24.
Single-molecule fluorescence resonance energy transfer (smFRET) is a powerful tool to investigate the dynamics of biomolecular events in real time. However, it requires two fluorophores and can be applied only to dynamics that accompany large changes in distance between the molecules. Herein, we introduce a method for kinetic analysis based on control of fluorescence blinking (KACB), a general approach to investigate the dynamics of biomolecules by using a single fluorophore. By controlling the kinetics of the redox reaction the blinking kinetics or pattern can be controlled to be affected by microenvironmental changes around a fluorophore (rKACB), thereby enabling real-time single-molecule measurement of the structure-changing dynamics of nucleic acids.
单分子荧光共振能量转移（smFRET）是实时研究生物分子事件动力学的有力工具。然而，它需要两个荧光团，并且只能应用于伴随分子间距离发生大变化的动力学。在此，我们介绍了一种基于荧光闪烁控制的动力学分析方法（KACB），这是一种使用单个荧光团研究生物分子动力学的通用方法。通过控制氧化还原反应的动力学，可以控制闪烁动力学或模式受到荧光团周围微环境变化的影响（rKACB），从而实现核酸结构变化动力学的实时单分子测量。
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