Affiliations: Dipartimento di Fisica e Matematica and CNISM–CNR, Universita' degli Studi dell'Insubria, Como, Italy | National Laboratory for Ultrafast and Ultraintense Optical Science – CNR–INFM, Como, Italy
Note: [] Corresponding author: Luca Nardo, Dipartimento di Fisica e Matematica, Universita' degli Studi dell'Insubria and CNISM–CNR, Via Valleggio 11, 22100 Como, Italy. Tel.: +39 0312 386254; Fax: +39 0312 386119; E-mail: [email protected].
Abstract: We perform time-correlated single-photon counting measurements with 30-ps resolution of the fluorescence emitted by the donor fluorophores of donor–acceptor pairs covalently labeling DNA synthetic oligonucleotides by using a non-commercial single-photon avalanche diode. The measurements allow us to precisely assess the fluorescence resonant energy transfer efficiency of the pairs in solutions containing the oligonucleotides and DNA–ligands, for different ratios of the DNA base-pair to ligand concentrations. A quantitative evaluation of the deformations of DNA double strands following the ligand binding is obtained, as the transfer efficiency is a steep function of the donor-to-acceptor distance. The results lead to an easy and cheap method to discriminate between the binding modes of minor groove and base intercalating ligands.
Keywords: Binding-induced conformational transition, DNA base intercalator, DNA minor groove-binder, time-correlated single-photon counting, fluorescence resonance energy transfer
DOI: 10.3233/SPE-2009-0370
Journal: Spectroscopy, vol. 23, no. 1, pp. 11-28, 2009
