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SH2 Domains pp 291–305Cite as

Calorimetric Measurement of SH2 Domain Ligand Affinities

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1555))

Abstract

Isothermal titration calorimetry (ITC) has emerged as a leading approach in the characterization of protein/ligand interactions. This technique measures the heat change of a system upon binding of a ligand to a biomolecule, and thereby requires no immobilization, intrinsic fluorescence, or labeling of any kind of either species. If properly designed, a single experiment can not only measure the binding affinity, but also determine additional binding and thermodynamic parameters, including the enthalpy, entropy, and the stoichiometry of the interaction. Here, we describe the protocol for the collection of calorimetric data for the binding of peptides to SH2 protein domains.

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Acknowledgments

We would like to thank Dr. Annette Erbse for her generous feedback and technical support. We would also like to thank our collaborators Dr. Zhongping Tan and Dr. Xiaoyang Guan for the phosphopeptide synthesis, as well as Dr. Julie Forman-Kay for kindly providing us with a PLCγ1-pET11d construct. This work was supported by NSF grant MCB1121842 (to D.S.W.).

Author information

Authors and Affiliations

  1. Department of Chemistry and Biochemistry, University of Colorado, 215 UCB, Boulder, CO, 80309, USA

    Marissa A. McKercher & Deborah S. Wuttke

Authors
  1. Marissa A. McKercher
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  2. Deborah S. Wuttke
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Corresponding author

Correspondence to Deborah S. Wuttke .

Editor information

Editors and Affiliations

  1. Department of Genetics and Genome Sciences, University of Connecticut School of Medicine, Farmington, Connecticut, USA

    Kazuya Machida

  2. Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA

    Bernard A. Liu

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McKercher, M.A., Wuttke, D.S. (2017). Calorimetric Measurement of SH2 Domain Ligand Affinities. In: Machida, K., Liu, B. (eds) SH2 Domains. Methods in Molecular Biology, vol 1555. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6762-9_16

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  • DOI: https://doi.org/10.1007/978-1-4939-6762-9_16

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6760-5

  • Online ISBN: 978-1-4939-6762-9

  • eBook Packages: Springer Protocols

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