Characterization of a human liver cytochrome P-450 involved in the oxidation of debrisoquine and other drugs by using antibodies raised to the analogous rat enzyme.
Debrisoquine 4-hydroxylase activity is a prototype for genetic polymorphism in oxidative drug metabolism in humans; approximately 10% of Caucasian populations exhibit the poor metabolizer phenotype, and the clearance of at least 14 other drugs has been shown to be deficient in patients exhibiting this phenotype. Antibodies prepared to a cytochrome P-450 shown to be responsible for debrisoquine 4-hydroxylation in rats were found to inhibit the oxidation of debrisoquine and sparteine, encainide, and propranolol, three other drugs suggested to be associated with this phenotype, in human liver microsomes. The antibodies did not inhibit the oxidation of seven other cytochrome P-450 substrates. The antibodies recognized a single polypeptide of Mr51,000 after combined sodium dodecyl sulfate/polyacrylamide electrophoresis and immunochemical staining of human liver microsomes. The intensity of this band was significantly correlated with debrisoquine 4-hydroxylase activity when liver microsomes from 44 organ donors were examined. Immunoprecipitation of in vitro translation products of total liver RNA revealed major electrophoretic bands corresponding to the cytochrome P-450 in rats and humans. The level of translatable mRNA coding for the debrisoquine-hydroxylating cytochrome P-450 was an order of magnitude less in human liver than in rat liver. The availability of these antibodies provides a biochemical basis for further basic and clinical studies on the role of a particular cytochrome P-450 polymorphism in humans.
D000818:Animals; D000906:Antibodies; D000936:Antigen-Antibody Complex; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D003647:Debrisoquin / Q000378:metabolism*; D006801:Humans; D007122:Immunoelectrophoresis; D007074:Immunoglobulin G; D007546:Isoquinolines / Q000378:metabolism*; D007700:Kinetics; D008862:Microsomes, Liver / Q000378:metabolism*; D006899:Mixed Function Oxygenases / Q000037:antagonists & inhibitors; D010084:Oxidation-Reduction; D051381:Rats; D013045:Species Specificity
D000906:Antibodies; D000936:Antigen-Antibody Complex; D007074:Immunoglobulin G; D007546:Isoquinolines; D003577:Cytochrome P-450 Enzyme System; D006899:Mixed Function Oxygenases; D003647:Debrisoquin