Purified prostaglandin synthase activates aromatic amines to derivatives that are mutagenic to Salmonella typhimurium.
Prostaglandin H synthase (PHS) is widely distributed in mammalian tissues and has the ability to oxidize a variety of mutagens and carcinogens. It may therefore play a key role in the metabolic activation of xenobiotics. The present study documents that highly purified PHS can be used in conjunction with 5-phenyl-4-pentenyl-1-hydroperoxide (PPHP), a relatively stable and non-mutagenic hydroperoxide substrate, for the metabolic activation of aromatic amines to mutagenic derivatives that can be detected in short-term Salmonella typhimurium mutagenesis assays. The PHS-based activation system alone was not mutagenic for these tester strains, nor were the test compounds significantly toxic for the bacteria over the concentration range tested. When used in conjunction with Salmonella strains TA98 and TA100 in a modified Ames assay, this system should prove useful for screening of a wide range of compounds for metabolic activation by this mammalian peroxidase. The potential broad utility of this purified PHS-dependent metabolic activation system was investigated by evaluating the activation of 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) and 2-amino-3,4-dimethylimidazo[4,5-f]quinoline (MeIQ), which are representative of a group of mutagenic and carcinogenic heterocyclic arylamines to which humans are exposed via their diet. Both IQ and MeIQ were activated by PHS to potent mutagens and confirm the utility of the PPHP/PHS system for the activation of premutagens. Whereas the extent of activation of aromatic amines by S9-based systems is significantly greater than for the PHS activation system described herein, PHS may play a significant role in target tissues in which it is present at significantly greater levels than P450 isoenzymes. Moreover, it is likely that the substrate specificity of PHS differs sufficiently from that of P450 isoenzymes so that PHS may activate some compounds that are not efficiently activated by mixed-function oxidase based systems.
Sarkar
F H
FH
Oxford Biomedical Research, Inc., Rochester Hills, MI 48309.
Radcliff
G
G
Callewaert
D M
DM
D000475:Alkenes / Q000494:pharmacology*; D000588:Amines / Q000633:toxicity*; D001711:Biotransformation; D009152:Mutagenicity Tests; D009153:Mutagens; D010545:Peroxides / Q000494:pharmacology*; D011451:Prostaglandin-Endoperoxide Synthases / Q000302:isolation & purification / Q000494:pharmacology*; D012486:Salmonella typhimurium / Q000187:drug effects* / Q000235:genetics
D000475:Alkenes; D000588:Amines; D009153:Mutagens; D010545:Peroxides; C047254:5-phenylpent-4-enyl-1-hydroperoxide; D011451:Prostaglandin-Endoperoxide Synthases