Cytochrome c oxidase inhibition by anesthetics: thermodynamic analysis.
The thermodynamic parameters that characterize the inhibition of cytochrome c oxidase activity, in rat liver submitochondrial particles, by n-butanol, tetracaine, and dibucaine were obtained. Three equilibria were assumed in order to account for the data: for the interaction of inhibitor with the native state of the enzyme, for the interaction of inhibitor with the thermally (reversibly) denatured state, and for the change between the native and thermally denatured states. Inhibition results from interaction with both the native and denatured states but, because the interaction is stronger with the denatured than with the native state, the native/denatured equilibrium is shifted to the right by the anesthetics. The enthalpies of interaction are -2.3, -4.7, and 3.7 kcal/mol (1 cal = 4.18 J) for the native state and -10, -6, and -14 kcal/mol for the denatured state, for n-butanol, tetracaine, and dibucaine, respectively. These values are much smaller than the previous estimates obtained by using the assumption that anesthetics interact only with the thermally denatured state of enzymes (e.g., -81 kcal/mol for tetracaine inhibition of luciferase). Our results suggest that local anesthetics inhibit enzyme activity by causing a reversible perturbation of protein conformation. The magnitude of the perturbation is much smaller (in energetic terms) than that which accompanies thermal denaturation.
D000440:Butanols / Q000494:pharmacology; D003992:Dibucaine / Q000494:pharmacology; D003576:Electron Transport Complex IV / Q000037:antagonists & inhibitors*; D008930:Mitochondria, Liver / Q000201:enzymology; D011487:Protein Conformation; D013696:Temperature; D013748:Tetracaine / Q000494:pharmacology; D013816:Thermodynamics
D000440:Butanols; D013748:Tetracaine; D003576:Electron Transport Complex IV; D003992:Dibucaine