Isothermal Titration Calorimetry: Assisted Crystallization of RNA-Ligand Complexes.
Fiche publication
Date publication
janvier 2016
Journal
Methods in molecular biology (Clifton, N.J.)
Auteurs
Membres identifiés du Cancéropôle Est :
Dr ENNIFAR Eric
Tous les auteurs :
Da Veiga C, Mezher J, Dumas P, Ennifar E
Lien Pubmed
Résumé
The success rate of nucleic acids/ligands co-crystallization can be significantly improved by performing preliminary biophysical analyses. Among suitable biophysical approaches, isothermal titration calorimetry (ITC) is certainly a method of choice. ITC can be used in a wide range of experimental conditions to monitor in real time the formation of the RNA- or DNA-ligand complex, with the advantage of providing in addition the complete binding profile of the interaction. Following the ITC experiment, the complex is ready to be concentrated for crystallization trials. This chapter describes a detailed experimental protocol for using ITC as a tool for monitoring RNA/small molecule binding, followed by co-crystallization.
Mots clés
5' Untranslated Regions, Aminoglycosides, chemistry, Base Sequence, Calorimetry, methods, Crystallization, Crystallography, X-Ray, DNA, chemistry, Kinetics, Ligands, Molecular Sequence Data, Nucleic Acid Conformation, Paromomycin, analogs & derivatives, Protein Binding, RNA, chemistry, Software, Temperature, Thermodynamics
Référence
Methods Mol. Biol.. 2016 ;1320:127-43