Measurement of Biochemical Affinities with a Gill Titration Calorimeter
| Autor: | Mohamed El Harrous, Antonio Parody-Morreale |
|---|---|
| Rok vydání: | 1997 |
| Předmět: |
Enthalpy
Monte Carlo method Biophysics Analytical chemistry Calorimetry Biochemistry Cytidine Monophosphate Trypsin Enzyme Inhibitors Molecular Biology Equilibrium constant Plant Proteins Binding Sites Chemistry Titrimetry Ribonuclease Pancreatic Cell Biology Enzymes Calorimeter Kinetics Reagent Thermodynamics Physical chemistry Titration alpha-Amylases Trypsin Inhibitors Monte Carlo Method Protein Binding Macromolecule Dimensionless quantity |
| Zdroj: | Analytical Biochemistry. 254:96-108 |
| ISSN: | 0003-2697 |
| DOI: | 10.1006/abio.1997.2386 |
| Popis: | A Gill titration calorimeter is evaluated as an instrument to determine in one experiment the equilibrium constant and the enthalpy change of a biochemical reaction. The dimensionless parameter kc (the product of the association equilibrium constant and the concentration of the reagent to be titrated; Wiseman et al., Anal. Biochem. 179, 131-137, 1989) is used to analyze the instrument performance. The analysis of simulated titration data corresponding to a simple model case shows that association equilibrium constants in the 10(2)-10(7) M-1 range may be determined when the kc parameter is between 1 and 1000. In addition we use a Monte Carlo approach to estimate the precision in the thermodynamic parameters of the reaction under study. The relative precision in the calculated constants ranges from 3 to 80% depending on the macromolecule concentration and kc value in the experiment. These results were checked with the study of the reactions of beta-trypsin with its inhibitor and ribonuclease A with cytidine 2'-monophosphate and cytidine 3'-monophosphate. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
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