Xenobiotic-metabolizing cytochromes P450 convert prostaglandin endoperoxide to hydroxyheptadecatrienoic acid and the mutagen, malondialdehyde.
Cyclooxygenases catalyze the oxygenation of arachidonic acid to prostaglandin endoperoxides. Cyclooxygenase-2- and the xenobiotic-metabolizing cytochrome P450s 1A and 3A are all aberrantly expressed during colorectal carcinogenesis. To probe for a role of P450s in prostaglandin endoperoxide metabolism, we studied the 12-hydroxyheptadecatrienoate (HHT)/malondialdehyde (MDA) synthase activity of human liver microsomes and purified P450s. We found that human liver microsomes have HHT/MDA synthase activity that is concentration-dependent and inhibited by the P450 inhibitors, ketoconazole and clotrimazole with IC(50) values of 1 and 0.4 microM, respectively. This activity does not require P450 reductase. HHT/MDA synthase activity was present in purified P450s but not in heme alone or other heme proteins. The catalytic activities of various purified P450s were determined by measuring rates of MDA production from prostaglandin endoperoxide. At 50 microM substrate, the catalytic activities of purified human P450s varied from 10 +/- 1 to 0.62 +/- 0.02 min(-1), 3A4 >> 2E1 > 1A2. Oxabicycloheptane analogs of prostaglandin endoperoxide, U-44069 and U-46619, induced spectral changes in human P450 3A4 with K(s) values of 240 +/- 20 and 130 +/- 10 microM, respectively. These results suggest that co-expression of cyclooxygenase-2 and P450s in developing cancers may contribute to genomic instability due to production of the endogenous mutagen, MDA.
Plastaras
J P
JP
A. B. Hancock Jr. Memorial Laboratory for Cancer Research, Department of Biochemistry, Center in Molecular Toxicology and The Vanderbilt-Ingram Cancer Center, Vanderbilt University School of Medicine, Nashville, Tennessee 37232, USA.
Guengerich
F P
FP
Nebert
D W
DW
Marnett
L J
LJ
D019796:15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid / Q000378:metabolism; D000818:Animals; D002384:Catalysis; D002478:Cells, Cultured; D002855:Chromatography, Thin Layer; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D005231:Fatty Acids, Unsaturated / Q000378:metabolism*; D006418:Heme / Q000378:metabolism; D006801:Humans; D008315:Malondialdehyde / Q000378:metabolism*; D008862:Microsomes, Liver / Q000201:enzymology; D009153:Mutagens / Q000378:metabolism*; D011449:Prostaglandin Endoperoxides / Q000378:metabolism*; D044262:Prostaglandin H2; D011463:Prostaglandins H / Q000737:chemistry / Q000378:metabolism; D011817:Rabbits
D005231:Fatty Acids, Unsaturated; D009153:Mutagens; D011449:Prostaglandin Endoperoxides; D011463:Prostaglandins H; D044262:Prostaglandin H2; D006418:Heme; D008315:Malondialdehyde; C008033:12-hydroxy-5,8,10-heptadecatrienoic acid; D019796:15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid; D003577:Cytochrome P-450 Enzyme System