title: Isothermal titration calorimetry.
abstract: In the last two decades, isothermal titration calorimetry (ITC) has become the preferred technique to determine the binding energetics of biological processes, including protein-ligand binding, protein-protein binding, DNA-protein binding, protein-carbohydrate binding, protein-lipid binding, and antigen-antibody binding. In this unit several protocols are presented, ranging from the basic ones that are aimed at characterizing binding of moderate affinity to advanced protocols that are aimed at determining very high or very low affinity binding processes. Also, alternate protocols for special cases (homodimeric proteins and unstable proteins) and additional information accessible by ITC (heat capacity and protonation/deprotonation processes coupled to binding) are presented.
authors.affiliation: Johns Hopkins University, Baltimore, Maryland, USA.
authors.forename: Adrián
authors.initials: A
authors.lastname: Velázquez-Campoy
authors.forename: Hiroyasu
authors.initials: H
authors.lastname: Ohtaka
authors.forename: Azin
authors.initials: A
authors.lastname: Nezami
authors.forename: Salman
authors.initials: S
authors.lastname: Muzammil
authors.forename: Ernesto
authors.initials: E
authors.lastname: Freire
mesh_terms: D002151:Calorimetry / Q000295:instrumentation / Q000379:methods*; D019281:Dimerization; D008024:Ligands; D008954:Models, Biological; D046912:Multiprotein Complexes / Q000378:metabolism; D011485:Protein Binding; D011489:Protein Denaturation / Q000502:physiology; D011506:Proteins / Q000378:metabolism; D011522:Protons; D013223:Statistics as Topic; D013379:Substrate Specificity; D013696:Temperature; D017186:Titrimetry / Q000379:methods*
chemical_list: D008024:Ligands; D046912:Multiprotein Complexes; D011506:Proteins; D011522:Protons