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kinITC: A New Method for Obtaining Joint Thermodynamic and Kinetic Data by Isothermal Titration Calorimetry
- Burnouf, Dominique, Ennifar, Eric, Guedich, Sondes, Puffer, Barbara, Hoffmann, Guillaume, Bec, Guillaume, Disdier, François, Baltzinger, Mireille, Dumas, Philippe
- Journal of the American Chemical Society 2012 v.134 no.1 pp. 559-565
- Human immunodeficiency virus 1, RNA folding, RNA-directed DNA polymerase, calorimetry, ligands, processing technology, thermodynamics, titration
- Isothermal titration calorimetry (ITC) is the method of choice for obtaining thermodynamic data on a great variety of systems. Here we show that modern ITC apparatus and new processing methods allow researchers to obtain a complete kinetic description of systems more diverse than previously thought, ranging from simple ligand binding to complex RNA folding. We illustrate these new features with a simple case (HIV-1 reverse transcriptase/inhibitor interaction) and with the more complex case of the folding of a riboswitch triggered by the binding of its ligand. The originality of the new kinITC method lies in its ability to dissect, both thermodynamically and kinetically, the two components: primary ligand binding and subsequent RNA folding. We are not aware of another single method that can yield, in a simple way, such deep insight into a composite process. Our study also rationalizes common observations from daily ITC use.