Catalytic properties of CYP1A isoforms in the liver of an agnathan (Lampetra fluviatilis) and two species of teleost (Pleuronectes flesus, Anguilla anguilla).
The catalytic activity of CYP1A isoforms and the effect of mammalian CYP1A-specific inhibitors in liver S9 fractions were studied in an agnathan (River lamprey, Lampetra fluviatilis, 30-33 cm) and in two species of teleost fish (European flounder, Pleuronectes flesus, 11-18 cm and common eel, Anguilla anguilla, 31-48 cm). Ethoxyresorufin O-deethylation (EROD), caffeine N-demethylation/C-oxidation and phenacetin O-deethylation (POD) activity increased 3-4-fold in flounders and 17-46-fold in eels, 5 days after fish were injected (i.p.) with 100 mg kg(-1) benzo(a)pyrene (B[a]P). In lampreys, basal EROD activity was very low and no increase in activity was observed following exposure to B[a]P. While the apparent Michaelis constant (K(m)) for each assay showed only small changes after B[a]P injection, maximum reaction velocity (V(max)) values increased by up to 19- and 84-fold for EROD activity, 4- and 35-fold for caffeine-related metabolism and 4- and 19-fold for POD activity in flounders and eels, respectively. The mammalian CYP1A2 inhibitor furafylline (50 microM-1 mM) reduced activity in the EROD, caffeine and POD assays to 65, 21 and 20% of control values in flounders and to 85, 10 and 5% of control values in eels, respectively. By contrast, low concentrations (0.025-0.050 microM) of the mammalian CYP1A1 inhibitor ellipticine completely abolished EROD activity, but had no effect (up to 1 mM) on caffeine metabolism or POD activity in either species. While the inhibitor studies strongly suggest that two separate enzymes are present in flounders and eels, the monophasic Michaelis-Menten kinetics obtained in all the assays imply that only a single CYP1A protein is present that has substrate and inhibitor specificities characteristic of both mammalian CYP1A1 and CYP1A2 isoforms.
Rotchell
J M
JM
Department of Biology, Johns Hopkins University, 3400 North Charles Street, Baltimore MD 21113, USA.
Steventon
G B
GB
Bird
D J
DJ
D000811:Anguilla / Q000502:physiology*; D000818:Animals; D002110:Caffeine / Q000008:administration & dosage / Q000378:metabolism; D000697:Central Nervous System Stimulants / Q000008:administration & dosage / Q000378:metabolism; D019363:Cytochrome P-450 CYP1A1 / Q000737:chemistry / Q000378:metabolism*; D005432:Flounder / Q000502:physiology*; D007527:Isoenzymes; D007700:Kinetics; D007798:Lampreys / Q000502:physiology*; D008099:Liver / Q000201:enzymology; D010084:Oxidation-Reduction
D000697:Central Nervous System Stimulants; D007527:Isoenzymes; D002110:Caffeine; D019363:Cytochrome P-450 CYP1A1