Modulating Fluorescence Anisotropy of Terminally Labeled Double-Stranded DNA via the Interaction between Dye and Nucleotides for Rational Design of DNA Recognition Based Applications
Modulating Fluorescence Anisotropy of Terminally Labeled Double-Stranded DNA via the Interaction between Dye and Nucleotides for Rational Design of DNA Recognition Based Applications
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通过染料和核苷酸之间的相互作用调节末端标记双链 DNA 的荧光各向异性，以合理设计基于 DNA 识别的应用
DOI：
10.1021/ac504028n
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发表时间：
2015-03-03
影响因子：
7.4
通讯作者：
Li, Na
中科院分区：
文献类型：
--
作者：
Huang, Hongduan;Wei, Hejia;Li, Na
对于荧光各向异性方法的发展来说，以一种简单的方式有效地增强荧光各向异性是最理想的。这项工作旨在深入了解末端标记双链DNA(DsDNA)的荧光各向异性，以便为基于DNA识别的应用提供一种简单而通用的设计策略。我们证明了dsDNA的荧光各向异性可以由染料的性质、分子体积和dsDNA的末端结构来调节。荧光各向异性随着碱基对数目的增加而上升，直至18bp，然后趋于平稳，表明分子体积不是导致荧光各向异性的唯一因素。通过选择带正电中心的染料，由于静电相互作用限制了染料的节段性运动，获得了高的荧光各向异性信号。通过合理设计dsDNA的末端结构，在二维(2D)H-1-H-1核Overhauser增强谱(NOESY)的支持下，通过增大有效总转动体积可以进一步改善荧光各向异性。随着末端标记dsDNA荧光各向异性的成功增强，通过传感从大分子(DNA和蛋白质)到小分子(可卡因)的主要类别的分析物，展示了简单和通用的设计。
Effective signal enhancement for fluorescence anisotropy in a simple manner is most desirable for fluorescence anisotropy method development. This work aimed to provide insights into the fluorescence anisotropy of terminally labeled double-stranded DNA (dsDNA) to facilitate a facile and universal design strategy for DNA recognition based applications. We demonstrated that fluorescence anisotropy of dsDNA could be regulated by the nature of dyes, the molecular volume, and the end structure of dsDNA. Fluorescence anisotropy ascended with the increased number of base pairs up to 18 bp and leveled off thereafter, indicating the molecular volume was not the only factor responsible for fluorescence anisotropy. By choosing dyes with the positively charged center, high fluorescence anisotropy signal was obtained due to the confinement of the segmental motion of dyes through the electrostatic interaction. By properly designing the end structure of dsDNA, fluorescence anisotropy could be further improved by enlarging the effective overall rotational volume, as supported by two-dimensional (2D) H-1-H-1 nuclear Overhauser enhancement spectroscopy (NOESY). With the successful enhancement of the fluorescence anisotropy for terminally labeled dsDNA, simple and universal designs were demonstrated by sensing of major classes of analytes from macromolecules (DNA and protein) to small molecules (cocaine).