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Characterization of Protein-Protein Interactions by Isothermal Titration Calorimetry

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Protein-Protein Interactions

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1278))

Abstract

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 α-chymotrypsin) protein associations by ITC.

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Acknowledgments

This work was supported by grants from the National Science Foundation (MCB0641252) and the National Institutes of Health (GM56550 and GM57144).

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Authors and Affiliations

  1. Institute of Biocomputation and Physics of Complex Systems (BIFI), Joint Unit IQFR-CSIC-BIFI, Universidad de Zaragoza, Zaragoza, Spain

    Adrian Velazquez-Campoy

  2. Department of Biochemistry and Molecular and Cell Biology, Universidad de Zaragoza, Zaragoza, 50018, Spain

    Adrian Velazquez-Campoy

  3. Fundacion ARAID, Government of Aragon, Zaragoza, Spain

    Adrian Velazquez-Campoy

  4. Gilead Sciences, Inc., Foster City, CA, 94404, USA

    Stephanie A. Leavitt

  5. Department of Biology, Johns Hopkins University, 3400 N. Charles Street, Baltimore, MD, 21218, USA

    Ernesto Freire

Authors
  1. Adrian Velazquez-Campoy
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  2. Stephanie A. Leavitt
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  3. Ernesto Freire
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Correspondence to Ernesto Freire .

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Editors and Affiliations

  1. Department of Pharmacology and Emory Chemical Biology Discovery Center, Emory University School of Medicine, Atlanta, Georgia, USA

    Cheryl L. Meyerkord

  2. Department of Pharmacology, Department of Hematology & Medical Oncology, and Emory Chemical Biology Discovery Center, Emory University School of Medicine, Atlanta, USA

    Haian Fu

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Velazquez-Campoy, A., Leavitt, S.A., Freire, E. (2015). Characterization of Protein-Protein Interactions by Isothermal Titration Calorimetry. In: Meyerkord, C., Fu, H. (eds) Protein-Protein Interactions. Methods in Molecular Biology, vol 1278. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2425-7_11

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  • DOI: https://doi.org/10.1007/978-1-4939-2425-7_11

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2424-0

  • Online ISBN: 978-1-4939-2425-7

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