Fluorescence Redox Blinking Adaptable to Structural Analysis of Nucleic Acids.
Fluorescence Redox Blinking Adaptable to Structural Analysis of Nucleic Acids.
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DOI：
10.1002/chem.201705668
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发表时间：
2018-05
期刊：
影响因子：
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通讯作者：
Takafumi Miyata;Naohiko Shimada;A. Maruyama;K. Kawai
中科院分区：
文献类型：
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作者：
Takafumi Miyata;Naohiko Shimada;A. Maruyama;K. Kawai
闪烁现象是单分子荧光测量所特有的。通过设计一种具有合适暗态寿命τOFF的荧光团，设计了一种基于荧光闪烁控制的动力学分析方法来研究生物分子的动力学。通过控制基于氧化还原反应的闪烁(RKACB)，在单分子水平上研究了RNA的构象动力学。然而，对rKACB合适的荧光分子知之甚少，rKACB的应用仅限于分析核酸的发夹和双链结构。在这项工作中，不同的荧光分子，包括Alexa 488，R6G，Tamra，ATTO 647N和ATTO 655，对rKACB进行了评估。此外，rKACB还适用于DNA/DNA和DNA/RNA核酸双链的识别和抗原-抗体相互作用的研究。通过改变氧化剂的大小，可以确定所分析生物分子的目标区域的溶剂可及性。
The phenomenon of blinking is unique to single-molecule fluorescence measurements. By designing a fluorophore with an appropriate dark-state lifetime τoff , a kinetic analysis based on the control of fluorescence blinking (KACB) was devised to investigate the dynamics of biomolecules. By controlling the redox-reaction-based blinking (rKACB), conformational dynamics of RNA at the single-molecule level was previously investigated. However, there is little knowledge about suitable fluorescent molecules for rKACB, and the application of rKACB has been limited to the analysis of hairpins and duplex structures of nucleic acids. In this work, various fluorescent molecules, including Alexa 488, R6G, TAMRA, ATTO 647N and ATTO 655, were evaluated for rKACB. Moreover, rKACB was adapted to the discrimination of DNA/DNA and DNA/RNA nucleic acid duplexes and investigation of antigen-antibody interactions. By changing the size of the oxidant, it was possible to determine the solvent accessibility of the target domain of the analyzed biomolecules.