Mutagen activation by cDNA-expressed P(1)450, P(3)450, and P450a.
cDNAs for rodent P(1)450, P(3)450, and P450a were expressed in the modified vaccinia virus-T7 RNA polymerase system. Each P450 exhibited its appropriate molecular weight and characteristic enzyme activity. Aryl hydrocarbon hydroxylase activity was catalyzed by P(1)450, acetanilide hydroxylase by P(3)450, and testosterone 7 alpha-hydroxylase by P450a. Ethoxycoumarin deethylase was exhibited by both P(1)450 and P(3)450. Each expressed P450 was also analyzed for its ability to activate 19 carcinogens of diverse classes to their mutagenic forms. Most notable was the activation of several polycyclic aromatic hydrocarbons by P1 and the activation of acetylaminofluorene, 4-aminobiphenyl, and several heterocyclic amine food pyrolysate products by P(3)450. P450a, in contrast, showed slight mutagen activation only toward N-hydroxy-2-acetyl aminofluorene. The vaccinia virus-T7 RNA polymerase system described here can express cDNAs for diverse forms of P450, each of which can then be characterized for substrate and product specificity and for mutagen activation.
Aoyama
T
T
Laboratory of Molecular Carcinogenesis, National Cancer Institute, Bethesda, Maryland.
Gonzalez
F J
FJ
Gelboin
H V
HV
D015660:7-Alkoxycoumarin O-Dealkylase / Q000235:genetics / Q000378:metabolism; D000818:Animals; D001189:Aryl Hydrocarbon Hydroxylases / Q000235:genetics / Q000378:metabolism; D001711:Biotransformation; D002273:Carcinogens / Q000378:metabolism; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics* / Q000378:metabolism; D004247:DNA / Q000187:drug effects / Q000235:genetics*; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D005822:Genetic Vectors; D009153:Mutagens / Q000378:metabolism*; D051381:Rats; D013250:Steroid Hydroxylases / Q000235:genetics / Q000378:metabolism; D013604:T-Phages / Q000201:enzymology; D014616:Vaccinia virus / Q000235:genetics
D002273:Carcinogens; D009153:Mutagens; D004247:DNA; D003577:Cytochrome P-450 Enzyme System; D013250:Steroid Hydroxylases; D015660:7-Alkoxycoumarin O-Dealkylase; D001189:Aryl Hydrocarbon Hydroxylases; C023738:acetanilide hydroxylase; C022724:testosterone 7-alpha-hydroxylase, hamster; D012321:DNA-Directed RNA Polymerases