Protocol

Drug-DNA Interaction Protocols

Volume 613 of the series Methods in Molecular Biology pp 25-35

Date:

Thermal Melting Studies of Ligand DNA Interactions

  • Aurore GuédinAffiliated withEquipe Santé, Laboratoire ‘Régulation et Dynamique des Génomes’, Muséum National d’Histoire Naturelle USM 503, INSERM UR 565, CNRS UMR 5153
  • , Laurent LacroixAffiliated withEquipe Santé, Laboratoire ‘Régulation et Dynamique des Génomes’, Muséum National d’Histoire Naturelle USM 503, INSERM UR 565, CNRS UMR 5153
  • , Jean-Louis MergnyAffiliated withEquipe Santé, Laboratoire ‘Régulation et Dynamique des Génomes’, Muséum National d’Histoire Naturelle USM 503, INSERM UR 565, CNRS UMR 5153 Email author 

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Abstract

A simple thermal melting experiment may be used to demonstrate the stabilization of a given structure by a ligand (usually a small molecule, sometimes a peptide). Preparation of the sample is straightforward, and the experiment itself requires an inexpensive apparatus. Furthermore, reasonably low amounts of sample are required. A qualitative analysis of the data is simple: An increase in the melting temperature (T m) indicates preferential binding to the folded form as compared to the unfolded form. However, it is perilous to derive an affinity constant from an increase in T m as other factors play a role.

Key words

FRET Telomeres Telomerase inhibitor G-quadruplex G-Quartet DNA ligands