Paketurytė, V, Zubrienė, A, Ladbury, JE orcid.org/0000-0002-6328-7200 et al. (1 more author) (2019) Intrinsic Thermodynamics of Protein-Ligand Binding by Isothermal Titration Calorimetry as Aid to Drug Design. In: Ennifar, E, (ed.) Microcalorimetry of Biological Molecules: Methods and Protocols. Methods in Molecular Biology, 1964 . Springer (Humana Press) , New York, N.Y., U.S.A. , pp. 61-74. ISBN 978-1-4939-9178-5
Abstract
Isothermal titration calorimetry (ITC) is one of the main techniques to determine specific interactions between molecules dissolved in aqueous solution. This technique is commonly used in drug development programs when low-molecular-weight molecules are sought that bind tightly and specifically to a protein (disease target) molecule. The method allows a complete thermodynamic characterization of an interaction, i.e., ITC enables direct determination of the model-independent observed interaction change in enthalpy (ΔH) and a model-dependent observed interaction affinity (change in Gibbs free energy, ΔG) in a single experiment. The product of temperature and change in entropy (TΔS) can be obtained by the subtraction of ΔG from ΔH, and the change in heat capacity (ΔC p ) can be determined as a slope of the temperature dependence of the binding ΔH. Despite the apparent value of ITC in characterization of interactions, it is often forgotten that many protein-ligand binding reactions are linked to protonation-deprotonation reactions or various conformational changes. In such cases, it is important to determine the linked-reaction contributions and obtain the intrinsic values of the changes in Gibbs energy (affinity), enthalpy, and entropy. These energy values can then be used in various SAR-type structure-thermodynamics and combined with structure-kinetics correlations in drug design, when searching for small molecules that would bind the protein target molecule. This manuscript provides a detailed protocol on how to determine the intrinsic values of protein-ligand binding thermodynamics by ITC.
Metadata
Item Type: | Book Section |
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Authors/Creators: |
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Editors: |
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Copyright, Publisher and Additional Information: | (c) 2019, Springer Science+Business Media, LLC, part of Springer Nature. This is an author produced version of a chapter published in Microcalorimetry of Biological Molecules: Methods and Protocols. Uploaded in accordance with the publisher's self-archiving policy. |
Keywords: | Isothermal titration calorimetry; ITC; Enthalpy of binding; Gibbs energy of binding; Drug design; Intrinsic thermodynamics of binding |
Dates: |
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Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Biological Sciences (Leeds) > School of Molecular and Cellular Biology (Leeds) |
Depositing User: | Symplectic Publications |
Date Deposited: | 23 May 2019 16:24 |
Last Modified: | 31 Mar 2020 00:38 |
Published Version: | https://link.springer.com/book/10.1007/978-1-4939-... |
Status: | Published |
Publisher: | Springer (Humana Press) |
Series Name: | Methods in Molecular Biology |
Identification Number: | 10.1007/978-1-4939-9179-2_5 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:146230 |