Immunochemical analysis of cytochromes P-450 in teleosts with polyclonal antibodies to cytochrome P-450E

Conference · Fri Feb 28 23:00:00 EST 1986 · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)

OSTI ID:5493239

Kloepper-Sams, P J; Park, S S; Gelboin, H V; Stegeman, J J

Cytochrome P-450E, an aryl hydrocarbon hydroxylase from liver microsomes of the marine fish Stenotomus chrysops (scup) served as antigen to produce monoclonal and polyclonal antibodies (MAb; PAb). Immuno- (western) blots with HRP-linked second Ab were used to examine the specificity of anti-P-450E, and levels of P-450E or its analogs in untreated and variously-treated scup and other teleosts. P-450E titrations were linear from a detection limit of 0.8 pmol to 7 pmol. PAb and MAb each recognized only P-450E and no other purified scup P-450 forms. The MAb recognized no microsomal proteins other than P-450E. P-450E comprised 40 to 60% of total liver microsomal P-450 in scup treated with methylcholanthrene (MC), ..beta..-naphthoflavone (BNF) or polychlorinated biphenyls, and less than 10% in controls. Feral scup showed a correlation between levels of inducible monooxygenase activity and immunodetectable P-450E. PAb and MAb each detected a single 58Kd band corresponding to the P-450E analog LM/sub 4b/ in MC-treated rainbow trout, and also detected single bands in BNF-induced brook trout and in winter flounder, indicating strong cross-reactivity. These probes will be sensitive tools in further studies of the function, regulation and phylogenetic relationships of inducible P-450s in freshwater and marine fish.

• Research Organization: Woods Hole Oceanographic Institute, MA
• OSTI ID: 5493239
• Report Number(s): CONF-8604222-
• Journal Information: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Journal Name: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States) Vol. 45:3; ISSN FEPRA
• Country of Publication: United States
• Language: English

Related Subjects

560305* -- Chemicals Metabolism & Toxicology-- Vertebrates-- (-1987)
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ANIMALS
ANTIBODIES
AQUATIC ORGANISMS
AROMATICS
BODY
CELL CONSTITUENTS
CHEMICAL ANALYSIS
CHLORINATED AROMATIC HYDROCARBONS
CHOLANTHRENE
CONDENSED AROMATICS
CYTOCHROMES
DIGESTIVE SYSTEM
ENZYME ACTIVITY
ENZYMES
FISHES
GLANDS
HALOGENATED AROMATIC HYDROCARBONS
HYDROXYLASES
LIVER
METABOLISM
MICROSOMES
MONOCLONAL ANTIBODIES
ORGANIC CHLORINE COMPOUNDS
ORGANIC COMPOUNDS
ORGANIC HALOGEN COMPOUNDS
ORGANOIDS
ORGANS
OXIDOREDUCTASES
OXYGENASES
PIGMENTS
PROTEINS
VERTEBRATES