Inhibition constants are typically measured using kinetic assays. These assays involve varying the concentrations of the substrate and inhibitor and measuring the reaction rate. The data are then analyzed using various models, such as the Michaelis-Menten equation, to determine the Ki values. Common methods include:
- Lineweaver-Burk Plot: A double-reciprocal plot of reaction rate data used to distinguish between types of inhibition and to estimate Ki. - Dixon Plot: A graphical representation of the inverse of reaction rates against inhibitor concentration, useful for determining Ki in competitive inhibition. - Direct Fitting of Progress Curves: Modern software tools can directly fit experimental data to enzyme kinetic models to provide Ki values without the need for linearization.
