title: Catalytic and immunochemical characterization of hepatic microsomal cytochromes P450 in beluga whale (Delphinapterus leucas).
abstract: Understanding the effects of environmental contaminants on cetaceans and other marine mammals will require information on the biochemistry of xenobiotic metabolism in these species. We characterized the hepatic microsomal cytochrome P450 system in beluga whales (Delphinapterus leucas) from the Canadian Arctic. The content of native P450 averaged 0.203 and 0.319 nmol/mg microsomal protein, cytochrome b5 content averaged 0.199 and 0.236 nmol/mg, and rates of NADPH-cytochrome c reductase were 79 and 76 nmol/min/mg, for females and males respectively. Ethoxyresorufin O-deethylase (EROD), pentoxyresorufin O-depentylase (PROD), and benzo[a]pyrene (BP) hydroxylase (AHH) activities were significantly greater in males than in females, and were highly correlated with one another (r2 between 0.853 and 0.912). HPLC analysis of in vitro BP metabolites revealed benzo-ring (7,8- and 9,10-) dihydrodiols, consistent with activation of this compound, as well as 4,5-dihydrodiol,3-OH-, 7-OH-, and 9-OH-BP and 1,6- and 3,6-quinones. Estradiol 2-hydroxylase activity did not differ between sexes, and rates did not correlate with those of the other activities. Antibodies against scup P450B (an apparent teleost CYP2B) and rat CYP2B1 did not recognize proteins in beluga liver microsomes, but there was a protein detected by antibodies to PB-inducible rabbit CYP2B4. Antibodies to ethanol and ketone-inducible rat CYP2E1 reacted with two proteins in beluga liver microsomes. Antibodies specific to hydrocarbon-inducible CYP1A1 and/or CYP1A2 forms showed a single protein band, apparently more closely related to CYP1A1. The content of CYP1A was fivefold greater in male than in female beluga. CYP1A content was highly correlated with EROD, PROD, and AHH activities, suggesting that this P450 form is a primary catalyst for these reactions in beluga. CYP1A content and activity were highly correlated with the concentrations in blubber of non-ortho and mono-ortho PCB congeners, compounds that induce CYP1A in other mammals. These results indicate that a CYP1A is a catalyst for the metabolism of aromatic hydrocarbon pollutants in the beluga whale, and strongly suggest that this protein is induced in these organisms by environmental contaminants, including PCBs. The results support the measurement of CYP1A expression as a biomarker of exposure to inducers in marine mammals. The full functional and evolutionary relationships of beluga CYP1A and of beluga proteins immunologically related to other P450 forms are uncertain.
authors.affiliation: Biology Department, Woods Hole Oceanographic Institution, Massachusetts 02543.
authors.forename: R D
authors.initials: RD
authors.lastname: White
authors.forename: M E
authors.initials: ME
authors.lastname: Hahn
authors.forename: W L
authors.initials: WL
authors.lastname: Lockhart
authors.forename: J J
authors.initials: JJ
authors.lastname: Stegeman
mesh_terms: D000818:Animals; D001564:Benzo(a)pyrene / Q000378:metabolism; D002384:Catalysis; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D015786:Cytochromes b5 / Q000378:metabolism; D004579:Electron Transport; D004790:Enzyme Induction; D005260:Female; D015151:Immunoblotting; D007527:Isoenzymes / Q000378:metabolism; D008297:Male; D008862:Microsomes, Liver / Q000201:enzymology*; D014907:Whales / Q000378:metabolism*
chemical_list: D007527:Isoenzymes; D001564:Benzo(a)pyrene; D015786:Cytochromes b5; D003577:Cytochrome P-450 Enzyme System