Isothermal Titration Calorimetry: Assisted Crystallization of RNA–Ligand Complexes

  • Cyrielle Da Veiga
  • Joelle Mezher
  • Philippe Dumas
  • Eric Ennifar
Part of the Methods in Molecular Biology book series (MIMB, volume 1320)

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.

Key words

Microcalorimetry Crystallization RNA–ligand complex ITC Thermodynamics 

Notes

Acknowledgments

This work was supported by grants from the Agence Nationale pour la Recherche (grant ANR-12-BS07-0007-03 “ClickEnARN”) and the Agence Nationale de Recherches sur le SIDA (ANRS). The authors would like to thank Vincent Olieric (Paul Scherrer Institute/Swiss Light Source, Villigen, Switzerland), Natalia Markova and Peter Gimeson (Microcal-Malvern, Uppsala, Sweden).

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Cyrielle Da Veiga
    • 1
  • Joelle Mezher
    • 1
  • Philippe Dumas
    • 1
  • Eric Ennifar
    • 1
  1. 1.Architecture et Réactivité de l’ARN, Institut de Biologie Moléculaire et CellulaireUPR 9002 CNRS/Université de StrasbourgStrasbourgFrance

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