Specificity of rabbit cytochrome P-450 isozymes involved in the metabolic activation of the food derived mutagen 2-amino-3-methylimidazo[4,5-f] quinoline.
The involvement of rabbit liver cytochromes P-450 in the activation of the food derived heterocyclic amine mutagen, 2-amino-3-methylimidazo[4,5-f]quinoline (IQ), was assessed using the Ames/Salmonella test. The number of revertants induced by IQ per microgram of control rabbit liver microsomes was 1872 +/- 50 (SD, n = 3), and this increased to 3690 +/- 239 when microsomes from 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) animals were used as the metabolic activation source. Microsomes from phenobarbital, rifampicin, and acetone pretreated rabbits were less efficient the controls at activating IQ to a mutagen. Cytochrome P-450 Forms 4 and 6, which are induced by TCDD, were found to be efficient activators of IQ to a mutagen in reconstitution experiments. Form 4 was 7.7-fold more active than Form 6 and produced 1702 revertants/0.125 pmol with a 20-min preincubation step in the Ames test. Anti-Form 4 IgG inhibited the activation of IQ in control and TCDD induced microsomes by 78 and 79%, respectively. The contents of cytochrome P-450 Form 4, determined by Western blot analysis, in control and phenobarbital, acetone, rifampicin, and TCDD pretreated microsomes were 0.55 +/- 0.19, 0.63 +/- 0.34, 0.5 +/- 0.27, 0.28 +/- 0.16, and 2.19 +/- 0.43 (n = 3) nmol/mg protein, respectively. A highly significant statistical correlation existed between the capacity of the above microsomes to activate IQ to a mutagen and their cytochrome P-450 Form 4 content (r = 0.96; r2 = 0.92). The content of cytochrome P-450 Form 6 in the above microsomes was also highly correlated with their capacity to activate IQ (r = 0.92; r2 = 0.85). Based on these results and the tissue distribution of cytochrome P-450 Forms 4 and 6, the former obviously contributes most toward the activation of IQ in the liver, whereas Form 6 would be expected to be primarily involved in this process in extrahepatic tissues.
McManus
M E
ME
Department of Clinical Pharmacology, School of Medicine, Flinders University of South Australia, Bedford Park.
Burgess
W
W
Snyderwine
E
E
Stupans
I
I
D000818:Animals; D001571:Benzoflavones / Q000494:pharmacology; D001711:Biotransformation; D003577:Cytochrome P-450 Enzyme System / Q000502:physiology*; D005260:Female; D007527:Isoenzymes / Q000502:physiology*; D008297:Male; D009153:Mutagens / Q000378:metabolism*; D010105:Oxygenases / Q000502:physiology; D000072317:Polychlorinated Dibenzodioxins / Q000494:pharmacology; D011804:Quinolines / Q000378:metabolism*; D011817:Rabbits; D019324:beta-Naphthoflavone
D001571:Benzoflavones; D007527:Isoenzymes; D009153:Mutagens; D000072317:Polychlorinated Dibenzodioxins; D011804:Quinolines; C029216:2-amino-3-methylimidazo(4,5-f)quinoline; D019324:beta-Naphthoflavone; D003577:Cytochrome P-450 Enzyme System; D010105:Oxygenases; C020066:dimethylaniline monooxygenase (N-oxide forming)