Survey of the year 2009: applications of isothermal titration calorimetry

Abstract

Isothermal titration calorimetry (ITC) is now an established and invaluable method for determining the thermodynamic constants, association constant and stoichiometry of molecular interactions in aqueous solutions. The technique has become widely used by biochemists to study protein interaction with other proteins, small molecules, metal ions, lipids, nucleic acids and carbohydrates; and nucleic acid interaction with small molecules. The drug discovery industry has utilized this approach to measure protein (or nucleic acid) interaction with drug candidates. ITC has been used to screen candidates, guide the design of potential drugs and validate the modelling used in structure-based drug design. Emerging disciplines including nanotechnology and drug delivery could benefit greatly from ITC in enhancing their understanding and control of nano-particle assembly, and drug binding and controlled release.

Thermodynamic Signatures of Macromolecular Complexes ‒ Insights on the Stability and Interactions of Nucleoplasmin, a Nuclear Chaperone

“…Extensive reviews have been published on the basic thermodynamic formalism, calorimeters' design and application of DSC [17][18][19][20] and ITC [18,[20][21][22][23][24]. Moreover, surveys on ITC application are published annually since 2002 [25][26][27][28]. Calorimetry on proteins in general will be briefly summarized here and examples for NP will be thoroughly reviewed.…”

“…Recently a protocol for novel application of the technique has been elaborated, in which ITC is used as a tracking tool, combined with chromatography, for identification of target protein in biomolecular mixture [77] and it has been suggested to be valuable when the target protein or ligand is unknown. References for the wide spectrum and examples of novel applications of ITC can be found in the surveys published each year in the Journal of Molecular Recognition [25][26][27][28].…”

Emerging methodologies to investigate lipid–protein interactions

“…40 Isothermal titration calorimetry using soluble lipid headgroup species has also been utilized, albeit less extensively. 41 To determine the ability of peripheral membrane proteins to insert into the hydrophobic core of the membrane the monolayer penetration technique 42 has been employed. 34b Additionally, the depth of penetration and protein orientation at the membrane interface have been determined by electron paramagnetic resonance spectroscopy.…”

Geometric features of the Wiseman isotherm in isothermal titration calorimetry

“…In particular, ITC is especially appropriate for studying protein-ligand, protein-protein, protein-nucleic acid, protein-membrane, ligand-membrane, and ligand-nucleic acid interactions (references [1][2][3][4][5][6][7], and references therein, provide an extensive, comprehensive, and varied inventory of experimental systems where ITC has been successfully employed in the last decade). ITC is the only technique that allows a complete binding characterization (simultaneous determination of the binding affinity, the binding enthalpy, and the stoichiometry) in a single experiment [8].…”