title: Modulating fluorescence anisotropy of terminally labeled double-stranded DNA via the interaction between dye and nucleotides for rational design of DNA recognition based applications.
abstract: 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) (1)H-(1)H 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).
authors: Beijing National Laboratory for Molecular Sciences, Key Laboratory of Bioorganic Chemistry and Molecular Engineering of Ministry of Education, Institute of Analytical Chemistry, College of Chemistry and Molecular Engineering, ‡Beijing NMR Center, and §College of Life Sciences, Peking University , Beijing, 100871, China.
authors: Hongduan
authors: H
authors: Huang
authors: Hejia
authors: H
authors: Wei
authors: Mingjian
authors: M
authors: Zou
authors: Xiao
authors: X
authors: Xu
authors: Bin
authors: B
authors: Xia
authors: Feng
authors: F
authors: Liu
authors: Na
authors: N
authors: Li
mesh_terms: D052157:Aptamers, Nucleotide / Q000032:analysis*; D003042:Cocaine / Q000032:analysis*; D004247:DNA / Q000737:chemistry* / Q000378:metabolism; D005454:Fluorescence Polarization / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D013917:Thrombin / Q000032:analysis*
chemical_list: D052157:Aptamers, Nucleotide; D005456:Fluorescent Dyes; D004247:DNA; D013917:Thrombin; D003042:Cocaine