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A dsDNA-lighted fluorophore for monitoring protein-ligand interaction through binding-mediated DNA protection

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Abstract

Because of their important roles in cellular functions, life activities, drug screening, and disease treatment, the development of efficient methods for monitoring protein-ligand interaction is essential. In this study, inspired by our previous studies on DNA conformation-selective fluorescent indicators, we developed a new sensing platform for monitoring protein-ligand interaction and detecting protein activity based on binding-mediated DNA protection and the dsDNA-lighted fluorophore, ethyl-4-[3,6-bis (1-methyl-4-vinylpyridium iodine)-9H-carbazol-9-yl)] butanoate (EBCB). The ligand was purposefully linked to the 3ʹ-terminal of a hairpin DNA probe to selectively bind with the target protein and protect the DNA from cleavage by exonuclease III. By virtue of EBCB’s outstanding capacity to discriminate DNA conformation, the protein-ligand interaction could be effectively monitored through a fluorescence change in EBCB. A high fluorescence signal was detected when the hairpin DNA was protected in the presence of the target protein, whereas a much lower signal was observed in the presence of nontarget proteins. Our results demonstrated that the proposed strategy had high potential, such as high selectivity and relatively high sensitivity, for monitoring protein-ligand interaction and detecting protein activity. We believe these results will pave the way for applying dsDNA-lighted fluorophore EBCB as an effective signal transducer for DNA conformation transformation-mediated biochemical sensing.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (21605008, 21735001, 21575018, 21505006) and the Hunan Provincial Natural Science Foundation (2016JJ3001).

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Authors and Affiliations

  1. Hunan Provincial Key Laboratory of Materials Protection for Electric Power and Transportation, Hunan Provincial Engineering Research Center for Food Processing of Aquatic Biotic Resources, School of Chemistry and Biological Engineering, Changsha University of Science and Technology, Changsha, 410114, China

    Zhihe Qing, Lixuan Zhu, Lina Hou, Zhen Zou, Sheng Yang & Ronghua Yang

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  1. Zhihe Qing
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  2. Lixuan Zhu
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  3. Lina Hou
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  4. Zhen Zou
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  5. Sheng Yang
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  6. Ronghua Yang
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Correspondence to Zhihe Qing or Ronghua Yang.

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Qing, Z., Zhu, L., Hou, L. et al. A dsDNA-lighted fluorophore for monitoring protein-ligand interaction through binding-mediated DNA protection. Sci. China Chem. 61, 1630–1636 (2018). https://doi.org/10.1007/s11426-018-9349-9

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  • DOI: https://doi.org/10.1007/s11426-018-9349-9

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