Abstract
Isothermal titration calorimetry (ITC) is a biophysical technique that allows a thermodynamic characterization of an interactive system. It is a free in solution technique that requires no labeling, using heat as signal. ITC allows simultaneous determination of affinity K a, stoichiometry n, enthalpy change ΔH and calculation of free energy change ΔG and entropy change ΔS in one single experiment. It is the only technique that allows direct enthalpy change measurement. By accessing the enthalpy change, we get a step closer in estimating the driving forces that characterize the interaction of a protein with a ligand, information much needed in the drug discovery process.
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References
Ladbury JE, Klebe G, Freire E (2010) Adding calorimetric data to decision making in lead discovery: a hot tip. Nat Rev Drug Discov 9:23–27
Atkins P, de Paula J (2002) Physical chemistry, 7th edn. Oxford University Press Inc., New York
Holdgate GA, Ward WHJ (2005) Measurements of binding thermodynamics in drug discovery. Drug Discov Today 10:1543–1550
Wiseman T et al (1989) Rapid measurement of binding constants and heats of binding using a new titration calorimeter. Anal Biochem 179:131–137
Ward WHJ, Holdgate GA (2001) Isothermal titration calorimetry in drug discovery. Prog Med Chem 38:309–376
Sturtevant JM (1977) Heat capacity and entropy changes in processes involving proteins. Proc Natl Acad Sci U S A 74:2236–2240
Niedzwiecka A et al (2002) Positive heat capacity change upon specific binding of translation initiation factor eIF4E to mRNA 5’cap. Biochemistry 41:12140–12148
Christensen JJ, Hansen LD, Izatt RM (1976) Handbook of proton ionization heats and related thermodynamic quantities. Wiley, New York
Fukada H, Takahashi K (1998) Enthalpy and heat capacity changes for the proton dissociation of various buffer components in 0.1 M potassium chloride. Proteins 33:159–166
Leavitt S, Freire E (2001) Direct measurement of protein binding energetics by isothermal titration calorimetry. Curr Opin Struct Biol 11:560–566
Jelesarov I, Bosshard HR (1999) Isothermal titration calorimetry and differential scanning calorimetry as complementary tools to investigate the energetics of biomolecular recognition. J Mol Recognit 12:3–18
Garidel P et al Understanding the self organisation of Association Colloids: Microcal Application Note. http://www.microcal.com/documents/colloidsappnote.pdf
Todd MJ, Gomez J (2001) Enzyme kinetics determined using calorimetry: a general assay for enzyme activity? Anal Biochem 296:179–187
Myszka DG et al (2003) The ABRF-MIRG’02 study: assembly state, thermodynamic and kinetic analysis of an enzyme/inhibitor interaction. J Biomol Tech 14:247–269
O'Brien R et al (1998) The effects of salt on the TATA binding protein-DNA interaction from a hyperthermophilic archaeon. J Mol Biol 279:117–125
Turnbull WB, Daranas AH (2003) On the value of c: can low affinity systems be studied by isothermal titration calorimetry? J Am Chem Soc 125:14859–14866
Velazquez-Campoy A, Freire E (2006) Isothermal titration calorimetry to determine association constants for high-affinity ligands. Nat Protoc 1:186–191
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Damian, L. (2013). Isothermal Titration Calorimetry for Studying Protein–Ligand Interactions. In: Williams, M., Daviter, T. (eds) Protein-Ligand Interactions. Methods in Molecular Biology, vol 1008. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-398-5_4
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DOI: https://doi.org/10.1007/978-1-62703-398-5_4
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