Cooking of meats produces several heterocyclic amines which are mutagenic and potentially carcinogenic. We found that metabolic activation of one of these heterocyclic amines, the quinoline derivative 2-amino-3-methylimidazo[4,5-f]quinoline (IQ), can be catalyzed by prostaglandin H synthase (PHS) as well as by CYP1A2. N-Acetyltransferase (NAT) increased IQ-DNA adduct formation by either of these pathways. In sonicate from transiently transfected COS cells, NAT1 increased CYP1A2 catalyzed adduct formation 4-fold while NAT2 increased adduct formation 12-fold. Both expressed human and purified ovine PHS-1 and PHS-2 catalyzed IQ-DNA adduct formation. The presence of NAT1 and NAT2 increased PHS-1 catalyzed adduct formation 2.5- and 4-fold, respectively. PHS-2 catalyzed IQ adduct formation was also enhanced by either NAT. The pyridine derivative, 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine, also produced by protein pyrolysis, did not form detectable DNA adducts during incubation with PHS. These results indicate that IQ is a substrate for both PHS-1 and PHS-2 and that NAT increases the ability of the resulting IQ metabolites to cause DNA damage. PHS activity, constitutive and induced, as well as NAT polymorphisms should be considered as factors in environmental carcinogenesis.