3 years ago

Synergetic Determination of Thermodynamic and Kinetic Signatures Using Isothermal Titration Calorimetry: A Full-Curve-Fitting Approach

Synergetic Determination of Thermodynamic and Kinetic Signatures Using Isothermal Titration Calorimetry: A Full-Curve-Fitting Approach
Lan Chen, Renxiao Liu, Ruimin Wang, Dexing Li, Guanglu Ge
Thermodynamic and kinetic signatures are pivotal information for revealing the binding mechanisms of biomolecules, and they play an indispensable role in drug discovery and optimization. While noncalorimetric methods measure only a part of these signatures, isothermal titration calorimetry (ITC) is considered to have the potential to acquire full signatures in an experiment. However, kinetic parameters are generally difficult to extract from ITC curves, as they are inevitably affected by the instrument-response function and the collateral heat of associated process during titrations. Thus, we herein report the development and validation of a full-curve-fitting method to resolve thermal power curves and to maximize the signal extraction using ITC. This method is then employed to quantify the dilution of an aqueous n-propanol solution and examine the inhibition of carbonic anhydrase by 4-carboxybenzenesulfonamide using a commercial instrument with a long apparent response time of ∼13 s.

Publisher URL: http://dx.doi.org/10.1021/acs.analchem.7b01091

DOI: 10.1021/acs.analchem.7b01091

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