Human cytochrome P450IIA3: cDNA sequence, role of the enzyme in the metabolic activation of promutagens, comparison to nitrosamine activation by human cytochrome P450IIE1.
We report that, in a human cell line, human cytochrome P450IIA3 is capable of metabolizing aflatoxin B1, benzo[a]-pyrene, N-nitrosodimethylamine (NDMA) and N-nitrosodiethylamine (NDEA) to cytotoxic and mutagenic species. Cytochrome P450IIA3-mediated activation of NDMA and NDEA was compared with human cytochrome P450IIE1-mediated activation in the same cell system. P450IIE1 was more effective at activating NDMA than P450IIA3, while P450IIA3 was more effective at activating NDEA than P450IIE1. Whole cells and microsomal fractions obtained from control cells and from cells expressing the P450IIA3 cDNA were characterized for expression of P450IIA3. Microsomal coumarin 7-hydroxylase activity was some 40 times greater in the transfected cells than in the control cells and was catalyzed by a protein that was immunochemically related to the rat liver cytochrome P450IIA gene family. Immunoblot analysis demonstrated that this protein was readily detectable in transfected cells but barely detectable in control cells. We also report the DNA and deduced amino acid sequence of the P450IIA3 cDNA isolate used in this study. Our isolate encodes a protein 489 amino acids that is five amino acids shorter at the N terminus but otherwise identical to a previously reported human P450IIA3 cDNA sequence.
Crespi
C L
CL
Gentest Corporation, Woburn, MA 01801.
Penman
B W
BW
Leakey
J A
JA
Arlotto
M P
MP
Stark
A
A
Parkinson
A
A
Turner
T
T
Steimel
D T
DT
Rudo
K
K
Davies
R L
RL
D016604:Aflatoxin B1; D000348:Aflatoxins / Q000378:metabolism; D001483:Base Sequence; D001564:Benzo(a)pyrene / Q000378:metabolism; D001711:Biotransformation; D003577:Cytochrome P-450 Enzyme System / Q000032:analysis / Q000235:genetics / Q000502:physiology*; D004247:DNA / Q000032:analysis*; D004052:Diethylnitrosamine / Q000378:metabolism*; D004128:Dimethylnitrosamine / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D009153:Mutagens / Q000378:metabolism*
D000348:Aflatoxins; D009153:Mutagens; D001564:Benzo(a)pyrene; D004052:Diethylnitrosamine; D004247:DNA; D003577:Cytochrome P-450 Enzyme System; D016604:Aflatoxin B1; D004128:Dimethylnitrosamine