Immunochemical analysis of cytochromes P-450 in teleosts with polyclonal antibodies to cytochrome P-450E
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), Vol. 45:3; Conference: 70. annual meeting of the Federation of American Society for Experimental Biology, St. Louis, MO, USA, 13 Apr 1986
- Country of Publication:
- United States
- Language:
- English
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CHLORINATED AROMATIC HYDROCARBONS
METABOLISM
CHOLANTHRENE
CYTOCHROMES
CHEMICAL ANALYSIS
OXYGENASES
ENZYME ACTIVITY
FISHES
HYDROXYLASES
LIVER
MICROSOMES
MONOCLONAL ANTIBODIES
ANIMALS
ANTIBODIES
AQUATIC ORGANISMS
AROMATICS
BODY
CELL CONSTITUENTS
CONDENSED AROMATICS
DIGESTIVE SYSTEM
ENZYMES
GLANDS
HALOGENATED AROMATIC HYDROCARBONS
ORGANIC CHLORINE COMPOUNDS
ORGANIC COMPOUNDS
ORGANIC HALOGEN COMPOUNDS
ORGANOIDS
ORGANS
OXIDOREDUCTASES
PIGMENTS
PROTEINS
VERTEBRATES
560305* - Chemicals Metabolism & Toxicology- Vertebrates- (-1987)