Applications of isothermal titration calorimetry in pure and applied research—survey of the literature from 2010

Abstract

Isothermal titration calorimetry (ITC) is a biophysical technique for measuring the formation and dissociation of molecular complexes and has become an invaluable tool in many branches of science from cell biology to food chemistry. By measuring the heat absorbed or released during bond formation, ITC provides accurate, rapid, and label-free measurement of the thermodynamics of molecular interactions. In this review, we survey the recent literature reporting the use of ITC and have highlighted a number of interesting studies that provide a flavour of the diverse systems to which ITC can be applied. These include measurements of protein-protein and protein-membrane interactions required for macromolecular assembly, analysis of enzyme kinetics, experimental validation of molecular dynamics simulations, and even in manufacturing applications such as food science. Some highlights include studies of the biological complex formed by Staphylococcus aureus enterotoxin C3 and the murine T-cell receptor, the mechanism of membrane association of the Parkinson's disease-associated protein α-synuclein, and the role of non-specific tannin-protein interactions in the quality of different beverages. Recent developments in automation are overcoming limitations on throughput imposed by previous manual procedures and promise to greatly extend usefulness of ITC in the future. We also attempt to impart some practical advice for getting the most out of ITC data for those researchers less familiar with the method.

Hot Biological Catalysis: Isothermal Titration Calorimetry to Characterize Enzymatic Reactions

“…In plants, the primary role of urease is to recycle nutrient nitrogen from arginase-derived urea 14 . Plant ureases are generally homohexamers a 6 with each subunit containing an active site with two Ni 2+ ions 15 . Among them, Canavalia ensiformis (jack bean) urease (JBU) has been the first protein to be crystallized in 1926 16 .…”

“…kinetics). In particular, isothermal titration calorimetry (ITC) has been adopted as method of choice to characterize the thermodynamics of biomolecular equilibria, involving protein-ligand, protein-protein, protein-metal ions and protein-DNA interactions [1][2][3][4][5][6] . In addition, the ability of ITC to provide kinetic information makes it a very powerful system to measure enzyme catalysis, although the potential of this application is still underestimated [7][8][9] .…”

How to Study Protein-protein Interactions

“…In addition, ITC can measure enzyme activity and kinetics, small molecule ?interactions and micelle formation. 58 While ITC is the best method for accurate quantitative measurements of interactions, one of the main drawbacks is a relatively large amount of sample needed for the experiment, in comparison with other biophysical approaches such as SPR or MST. However, the advent of the upgraded machines requiring significantly lower amounts of samples is gradually overcoming this problem.…”

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].…”