Characterization of protein-protein interactions by isothermal titration calorimetry

Adrian Velazquez-Campoy, Stephanie A. Leavitt, Ernesto Freire

Research output: Chapter in Book/Report/Conference proceedingChapter

8 Scopus citations


The analysis of protein-protein interactions has attracted the attention of many researchers from both a fundamental point of view and a practical point of view. From a fundamental point of view, the development of an understanding of the signaling events triggered by the interaction of two or more proteins provides key information to elucidate the functioning of many cell processes. From a practical point of view, understanding protein-protein interactions at a quantitative level provides the foundation for the development of antagonists or agonists of those interactions. Isothermal Titration Calorimetry (ITC) is the only technique with the capability of measuring not only binding affinity but the enthalpic and entropic components that define affinity. Over the years, isothermal titration calorimeters have evolved in sensitivity and accuracy. Today, TA Instruments and MicroCal market instruments with the performance required to evaluate protein-protein interactions. In this methods paper, we describe general procedures to analyze heterodimeric (porcine pancreatic trypsin binding to soybean trypsin inhibitor) and homodimeric (bovine pancreatic a-chymotrypsin) protein associations by ITC.

Original languageEnglish (US)
Title of host publicationProtein-Protein Interactions
Subtitle of host publicationMethods and Applications: Second Edition
PublisherSpringer New York
Number of pages22
ISBN (Electronic)9781493924257
ISBN (Print)9781493924240
StatePublished - Apr 9 2015


  • Binding
  • Calorimetry
  • Dimerization
  • Dissociation
  • Protein-protein interaction
  • Thermodynamics
  • Titration

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)


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