Isothermal titration calorimetry for drug design: Precision of the enthalpy and binding constant measurements and comparison of the instruments.
Anal Biochem
; 515: 61-64, 2016 Dec 15.
Article
en En
| MEDLINE
| ID: mdl-27717855
ABSTRACT
Isothermal titration calorimetry (ITC) is one of the most robust label- and immobilization-free techniques used to measure protein - small molecule interactions in drug design for the simultaneous determination of the binding affinity (ΔG) and the enthalpy (ΔH), both of which are important parameters for structure-thermodynamics correlations. It is important to evaluate the precision of the method and of various ITC instrument models by performing a single well-characterized reaction. The binding between carbonic anhydrase II and acetazolamide was measured by four ITC instruments - PEAQ-ITC, iTC200, VP-ITC, and MCS-ITC and the standard deviation of ΔG and ΔH was determined. Furthermore, the limit of an approach to reduce the protein concentration was studied for a high-affinity reaction (Kd = 0.3 nM), too tight to be measured by direct (non-displacement) ITC. Chemical validation of the enthalpy measurements is discussed.
Palabras clave
Texto completo:
1
Bases de datos:
MEDLINE
Asunto principal:
Termodinámica
/
Diseño de Fármacos
/
Anhidrasa Carbónica II
Límite:
Humans
Idioma:
En
Revista:
Anal Biochem
Año:
2016
Tipo del documento:
Article
País de afiliación:
Lituania