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Thioflavin T behaves as an efficient fluorescent ligand for label-free ATP aptasensor

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Abstract

Here, we for the first time demonstrated thioflavin T (ThT) as an efficient fluorescent ligand for 27-mer ATP-binding aptamer (ABA27), providing a novel signal readout mode for label-free selective ATP detection. ABA27 can promote the fluorescence emission of ThT with an unprecedentedly high efficiency, attributed to the specific structure of ABA27 rather than the G-tracts. Polyacrylamide gel electrophoresis, fluorescence spectroscopy, and fluorometric titration reveal that ThT interacts with ABA27 with a lower binding affinity (Kd ~89 μM) than ATP, which allows ATP to easily compete with ThT for the DNA binder. In the presence of ThT, adding ATP induces ABA27 to undergo a structural change, thereby not favoring the binding to ThT, verified by circular dichroism and UV-Vis absorption spectroscopy. As a result, the fluorescence intensity of ThT decreases dramatically, enabling the sensitive detection of ATP with high selectivity over other analogs. Such a sensing strategy may make ThT able to serve as a facile signal reporter for DNA nanomechanical devices fueled with ATP.

The principle of the displacement of ThT by ATP

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Acknowledgments

This work is supported by the National Natural Science Foundation of China (no. 21575133), National Key Research and Development Program (no. 2016YFA0201300), and the Recruitment Program of Global Experts.

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Authors and Affiliations

  1. Department of Chemistry, School of Chemistry and Materials Science, University of Science and Technology of China, Jinzhai Road 96, Hefei, Anhui, 230026, China

    Huihui Wang, Pai Peng, Shuangna Liu & Tao Li

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  1. Huihui Wang
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Correspondence to Tao Li.

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Wang, H., Peng, P., Liu, S. et al. Thioflavin T behaves as an efficient fluorescent ligand for label-free ATP aptasensor. Anal Bioanal Chem 408, 7927–7934 (2016). https://doi.org/10.1007/s00216-016-9926-9

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  • DOI: https://doi.org/10.1007/s00216-016-9926-9

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