Investigation of Anesthetic–Protein Interactions by a Thermodynamic Approach

Abstract Anesthetics can interact with a wide variety of proteins in the body, including ion channels and alter their activity, but little is known about the molecular mechanisms of the interactions responsible for the functional activity. Characterization of the nature of anesthetic–protein interactions therefore is important and requires the complete analysis of the binding energetics. Isothermal titration calorimetry (ITC) is the only technique that allows quantitative determination of all thermodynamic parameters, including the equilibrium binding constant ( K B ), the standard Gibbs free energy change (Δ G ), the enthalpy change (Δ H ), the entropy change (Δ S ), heat capacity change (Δ C p ), and stoichiometry ( n ) of the reaction. ITC does not require any labeling or modification of the interacting partners analyzed and can be performed in solution with small amounts of reagents. In this chapter we describe the general properties of the ITC method, highlighting some critical aspects of experimental planning and data analysis, with practical application to anesthetic–protein interactions.