Repeatability, precision, and accuracy of the enthalpies and Gibbs energies of a protein-ligand binding reaction measured by isothermal titration calorimetry.
Eur Biophys J
; 48(2): 139-152, 2019 Mar.
Article
en En
| MEDLINE
| ID: mdl-30535510
ABSTRACT
In rational drug design, it is important to determine accurately and with high precision the binding constant (the affinity or the change in Gibbs energy, ∆G), the change in enthalpy (ΔH), and the entropy change upon small molecule drug binding to a disease-related target protein. These thermodynamic parameters of the protein-ligand association reaction are usually determined by isothermal titration calorimetry (ITC). Here, the repeatability, precision, and accuracy of the measurement of the affinity and the change in enthalpy upon acetazolamide (AZM) interaction with human carbonic anhydrase II (CA II) are discussed based on the measurements using several ITC instruments. The AZM-CA II reaction was performed at decreasing protein-ligand concentrations until the determination of ∆G and ΔH was not possible, indicating a lower limit for accuracy. To obtain the confidence intervals (CI) of the ∆G and ΔH of AZM binding to CA II, the binding reaction was repeated numerous times at the optimal concentration of 10 µM and 25 °C temperature. The CI (at a confidence level α = 0.95) for ΔH = - 51.2 ± 1.0 kJ/mol and ∆G = - 45.4 ± 0.5 kJ/mol was determined by averaging the results of multiple repeats.
Palabras clave
Texto completo:
1
Bases de datos:
MEDLINE
Asunto principal:
Calorimetría
/
Anhidrasa Carbónica II
/
Acetazolamida
Límite:
Humans
Idioma:
En
Revista:
Eur Biophys J
Asunto de la revista:
BIOFISICA
Año:
2019
Tipo del documento:
Article
País de afiliación:
Lituania