Characterization of molecular interactions using isothermal titration calorimetry

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 1149; p. 193
• Main Authors: Krell, Tino, Lacal, Jesús, García-Fontana, Cristina, Silva-Jiménez, Hortencia, Rico-Jiménez, Miriam, Lugo, Andrés Corral, Darias, José Antonio Reyes, Ramos, Juan-Luis
• Format: Journal Article
• Language: English
• Published: United States 2014
• Subjects: Bacterial Proteins - metabolism; Buffers; Calorimetry - methods; Ligands; Protein Binding; Pseudomonas - metabolism; Signal-To-Noise Ratio; Statistics as Topic
• ISSN: 1940-6029
• DOI: 10.1007/978-1-4939-0473-0_16

Isothermal titration calorimetry (ITC) is based on a simple titration of one ligand with another and the small heat changes caused by the molecular interaction are detected. From one ITC experiment the complete set of thermodynamic parameters of binding including association and dissociation constants as well as changes in enthalpy, entropy, and free energy can be derived. Using this technique almost any type of molecular interaction can be analyzed. Both ligands are in solution, and there is no need for their chemical derivatization. There are no limits as to the choice of the analysis buffer, and the analysis temperature can be set between 4 and 80 °C. This technique has been primarily applied to study the interaction between various proteins of Pseudomonas with small molecule ligands. In addition, ITC has been used to study the binding of Pseudomonas proteins to target DNA fragments.