Trout P450IA1: cDNA and deduced protein sequence, expression in liver, and evolutionary significance.
The cytochrome P1(450) (P450IA1) cDNA has been isolated and sequenced from liver of 3-methylcholanthrene-treated rainbow trout (Salmo gairdneri). The cDNA hybridizes to a 2.8-kb mRNA that is induced at least 10-fold by 3-methylcholanthrene. Southern blot analysis suggests the presence of a single gene or a very small number of genes. An open reading frame of the 2573-bp cDNA encodes a 522-residue protein (Mr = 59,241) that is more similar to the mammalian P450IA1 than the mammalian P450IA2 proteins. The aromatic hydrocarbon (Ah) receptor, responsible for mammalian P450IA1 and IA2 inducibility, was detected in trout liver cytosol by specific binding to [1,6-3H]2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in vitro. Comparison of the fish P450IA1 protein with human, mouse, rat and rabbit P450IA1 and P450IA2 proteins reveals the presence of a remarkably large number of single amino acids and stretches of 2-6 residues in a row that are invariant among these nine P450 proteins. These conserved regions may participate in the binding of the NADPH-P450 oxidoreductase flavoprotein, substrate, or heme.
Heilmann
L J
LJ
USDA--Agricultural Research Service, Metabolism and Radiation Research Laboratory, Fargo, ND 58105.
Sheen
Y Y
YY
Bigelow
S W
SW
Nebert
D W
DW
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D005075:Biological Evolution; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics*; D004247:DNA / Q000235:genetics*; D005796:Genes; D006801:Humans; D008099:Liver / Q000187:drug effects / Q000378:metabolism*; D008748:Methylcholanthrene / Q000494:pharmacology; D008969:Molecular Sequence Data; D012487:Salmonidae / Q000235:genetics*; D012689:Sequence Homology, Nucleic Acid; D013045:Species Specificity; D014337:Trout / Q000235:genetics*
D008748:Methylcholanthrene; D004247:DNA; D003577:Cytochrome P-450 Enzyme System