Temperature dependence and mechanism of local anesthetic effects on mitochondrial adenosinetriphosphatase.
Chloroform-released ATPase prepared from beef heart mitochondria is inhibited by tetracaine and dibucaine over the entire temperature range in which the enzyme is active. The temperature of maximal activity is at 60 degrees C in the absence of anesthetic and is shifted upward by 2-3 degrees C by the addition of 0.3 mM dibucaine. Local anesthetics protect ATPase from irreversible cold inactivation. The kinetics of this protective effect are analyzed by a thermodynamic model in which the associated/dissociated subunit equilibrium is shifted toward the associated state by the preferential binding of anesthetic to the associated state. The accessibility of buried sulhydryl groups to reaction with 5,5'-dithiobis(2-nitrobenzoic acid) is increased by local anesthetics; this is interpreted to mean that the anesthetics increase the conformational flexibility of the protein. It is proposed that the hydrophobic moieties of local anesthetics and related compounds bind to numerous hydrophobic sites or crevices on ATPase; this binding induces a perturbation of the protein conformation, which in turn causes a decrease of enzyme activity. This model is sufficiently general to encompass the diversity of molecules which have similar anesthetic-like effects, and since it relates to common fundamental features of protein structure, it may also be the mechanism of the nonspecific effects of these molecules on other proteins.
D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000779:Anesthetics, Local / Q000494:pharmacology*; D000818:Animals; D002417:Cattle; D002725:Chloroform / Q000494:pharmacology; D002746:Chlorpromazine / Q000494:pharmacology; D003992:Dibucaine / Q000494:pharmacology; D004228:Dithionitrobenzoic Acid / Q000494:pharmacology; D007700:Kinetics; D008929:Mitochondria, Heart / Q000201:enzymology*; D013748:Tetracaine / Q000494:pharmacology; D013816:Thermodynamics
D000779:Anesthetics, Local; D013748:Tetracaine; D002725:Chloroform; D004228:Dithionitrobenzoic Acid; D000251:Adenosine Triphosphatases; D003992:Dibucaine; D002746:Chlorpromazine