title: Isothermal Titration Calorimetry: Assisted Crystallization of RNA-Ligand Complexes.
abstract: 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.
authors.affiliation: Architecture et Réactivité de l'ARN, Institut de Biologie Moléculaire et Cellulaire, UPR 9002 CNRS/Université de Strasbourg, 15 Rue René Descartes, Strasbourg, 67084, France.
authors.forename: Cyrielle
authors.initials: C
authors.lastname: Da Veiga
authors.forename: Joelle
authors.initials: J
authors.lastname: Mezher
authors.forename: Philippe
authors.initials: P
authors.lastname: Dumas
authors.forename: Eric
authors.initials: E
authors.lastname: Ennifar
mesh_terms: D020121:5' Untranslated Regions; D000617:Aminoglycosides / Q000737:chemistry; D001483:Base Sequence; D002151:Calorimetry / Q000379:methods*; D003460:Crystallization; D018360:Crystallography, X-Ray; D004247:DNA / Q000737:chemistry; D007700:Kinetics; D008024:Ligands; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010303:Paromomycin / Q000031:analogs & derivatives / Q000737:chemistry; D011485:Protein Binding; D012313:RNA / Q000737:chemistry*; D012984:Software; D013696:Temperature; D013816:Thermodynamics
chemical_list: D020121:5' Untranslated Regions; D000617:Aminoglycosides; D008024:Ligands; D010303:Paromomycin; D012313:RNA; D004247:DNA; C100306:lividomycin