Characterization of a common genetic defect of cytochrome P-450 function (debrisoquine-sparteine type polymorphism) — Increased michaelis constant (km) and loss of stereoselectivity of bufuralol 1′-hydroxylation in poor metabolizers
التفاصيل البيبلوغرافية
العنوان:
Characterization of a common genetic defect of cytochrome P-450 function (debrisoquine-sparteine type polymorphism) — Increased michaelis constant (km) and loss of stereoselectivity of bufuralol 1′-hydroxylation in poor metabolizers
Summary In order to define the mechanism of the debrisoquine-sparteine type genetic polymorphism of drug oxidation we studied the kinetics of bufuralol 1′-hydroxylation in liver microsomes from extensive and poor metabolizers and in a purified reconstituted human cytochrome P-450 isozyme with high activity for bufuralol 1′-hydroxylation, P-450[buf]. In extensive metabolizer microsomes the enzymatic reaction displayed apparent Michaelis-Menten kinetics and the (+)-isomer was preferentially metabolized. By contrast, the enzymatic reaction in poor metabolizer microsomes was characterized by a 4- to 5-fold increase in Km and by a loss of stereoselectivity. In a non-membraneous reconstituted system containing NADPH cytochrome P-450 reductase, a NADPH regenerating system and phospholipids, P-450[buf] exhibited an almost complete substrate stereoselectivity for (+)-isomer 1′- hydroxylation. It is concluded that the purified cytochrome P-450[buf] is the target of the debrisoquine-sparteine type oxidation polymorphism and that poor metabolizers have a quantitative or qualitative deficiency of this isozyme.
