Fluorescence Enhancement upon G-Quadruplex Folding: Synthesis, Structure, and Biophysical Characterization of a Dansyl/Cyclodextrin-Tagged Thrombin Binding Aptamer. - Archive ouverte HAL Access content directly
Journal Articles Bioconjugate Chemistry Year : 2013

Fluorescence Enhancement upon G-Quadruplex Folding: Synthesis, Structure, and Biophysical Characterization of a Dansyl/Cyclodextrin-Tagged Thrombin Binding Aptamer.

Abstract

A novel fluorescent thrombin binding aptamer (TBA), conjugated with the environmentally sensitive dansyl probe at the 3′-end and a β-cyclodextrin residue at the 5′-end, has been efficiently synthesized exploiting Cu(I)-catalyzed azide-alkyne cycloaddition procedures. Its conformation and stability in solution have been studied by an integrated approach, combining in-depth NMR, CD, fluorescence, and DSC studies. ITC measurements have allowed us to analyze in detail its interaction with human thrombin. All the collected data show that this bis-conjugated aptamer fully retains its G-quadruplex formation ability and thrombin recognition properties, with the terminal appendages only marginally interfering with the conformational behavior of TBA. Folding of this modified aptamer into the chairlike, antiparallel G-quadruplex structure, promoted by K+ and/or thrombin binding, typical of TBA, is associated with a net fluorescence enhancement, due to encapsulation of dansyl, attached at the 3′-end, into the apolar cavity of the β-cyclodextrin at the 5′-end. Overall, the structural characterization of this novel, bis-conjugated TBA fully demonstrates its potential as a diagnostic tool for thrombin recognition, also providing a useful basis for the design of suitable aptamer-based devices for theranostic applications, allowing simultaneously both detection and inhibition or modulation of the thrombin activity.
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Dates and versions

hal-00952351 , version 1 (26-02-2014)

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Stefano de Tito, François Morvan, Albert Meyer, Jean-Jacques Vasseur, Annunziata Cummaro, et al.. Fluorescence Enhancement upon G-Quadruplex Folding: Synthesis, Structure, and Biophysical Characterization of a Dansyl/Cyclodextrin-Tagged Thrombin Binding Aptamer.. Bioconjugate Chemistry, 2013, 24 (11), pp.1917-1927. ⟨10.1021/bc400352s⟩. ⟨hal-00952351⟩
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