Purification of the pyrazole-inducible cytochrome P-450 isozyme

Palakodety, R; Clejan, L; Krikun, G; Feierman, D; Cederbaum, A I

Title: Purification of the pyrazole-inducible cytochrome P-450 isozyme

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Abstract

The alcohol dehydrogenase inhibitor, pyrazole, appears to induce a cytochrome P-450 isozyme with properties similar to the ethanol-inducible P-450. The pyrazole-inducible P-450 isozyme was purified from the liver microsomes of rats treated with pyrazole essentially by the procedure of Ryan et al and also by chromatofocussing. The final preparation appeared homogenous by SDS-PAGE with an apparent molecular weight of 52,000, had a specific content of 11 nmoles P-450 per mg protein, showed very high activity of low K/sub m/ dimethylnitrosamine demethylase and produced a type II binding spectrum with dimethylsulfoxide. The enzyme was also active with aniline and aminopyrine as substrates. Pyrazole itself served as an excellent substrate with 4-hydroxy pyrazole being the product. An antibody against the pyrazole-inducible P-450 raised in chickens recognized a protein with mol.wt of about 52,000 in control microsomes. This band was highly enriched in microsomes from rats treated with pyrazole, 4-methyl-pyrazole, ethanol or acetone, but not phenobarbital or 3-methylcholanthrene. In summary, the pyrazole-inducible P-450 has been purified and appears to be identical in its catalytic and immunological properties to the alcohol-inducible P-450.

Authors:: Palakodety, R; Clejan, L; Krikun, G; Feierman, D; Cederbaum, A I

Publication Date:: Fri May 01 00:00:00 EDT 1987

Research Org.:: Mount Sinai School of Medicine, New York, NY

OSTI Identifier:: 6290676

Report Number(s):: CONF-870644-
Journal ID: CODEN: FEPRA

Resource Type:: Conference

Journal Name:: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)

Additional Journal Information:: Journal Volume: 46:6; Conference: 78. annual meeting of the American Society of Biological Chemists conference, Philadelphia, PA, USA, 7 Jun 1987

Country of Publication:: United States

Language:: English

Subject:: 63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.; 59 BASIC BIOLOGICAL SCIENCES; 3-METHYLCHOLANTHRENE; BIOLOGICAL EFFECTS; ACETONE; ETHANOL; MIXED-FUNCTION OXIDASES; BIOCHEMICAL REACTION KINETICS; FRACTIONATION; PYRAZOLES; DMSO; ELECTROPHORESIS; LIVER; MICROSOMES; MOLECULAR WEIGHT; PHENOBARBITAL; RATS; ALCOHOLS; ANESTHETICS; ANIMALS; ANTICONVULSANTS; AROMATICS; AZINES; AZOLES; BARBITURATES; BODY; CELL CONSTITUENTS; CENTRAL NERVOUS SYSTEM DEPRESSANTS; DIGESTIVE SYSTEM; DRUGS; ENZYMES; GLANDS; HETEROCYCLIC COMPOUNDS; HYDROCARBONS; HYDROXY COMPOUNDS; HYPNOTICS AND SEDATIVES; KETONES; KINETICS; MAMMALS; ORGANIC COMPOUNDS; ORGANIC NITROGEN COMPOUNDS; ORGANIC OXYGEN COMPOUNDS; ORGANIC SULFUR COMPOUNDS; ORGANOIDS; ORGANS; OXIDOREDUCTASES; OXYGENASES; POLYCYCLIC AROMATIC HYDROCARBONS; PYRIMIDINES; REACTION KINETICS; RODENTS; SEPARATION PROCESSES; SULFOXIDES; VERTEBRATES; 560300* - Chemicals Metabolism & Toxicology; 550200 - Biochemistry

Citation Formats


                    Palakodety, R, Clejan, L, Krikun, G, Feierman, D, and Cederbaum, A I. Purification of the pyrazole-inducible cytochrome P-450 isozyme.  United States: N. p., 1987. 
        Web.

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                    Palakodety, R, Clejan, L, Krikun, G, Feierman, D, & Cederbaum, A I. Purification of the pyrazole-inducible cytochrome P-450 isozyme.  United States.

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                    Palakodety, R, Clejan, L, Krikun, G, Feierman, D, and Cederbaum, A I. 1987.  
        "Purification of the pyrazole-inducible cytochrome P-450 isozyme".  United States.

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@article{osti_6290676,

  title        = {Purification of the pyrazole-inducible cytochrome P-450 isozyme},

  author       = {Palakodety, R and Clejan, L and Krikun, G and Feierman, D and Cederbaum, A I},

  abstractNote = {The alcohol dehydrogenase inhibitor, pyrazole, appears to induce a cytochrome P-450 isozyme with properties similar to the ethanol-inducible P-450. The pyrazole-inducible P-450 isozyme was purified from the liver microsomes of rats treated with pyrazole essentially by the procedure of Ryan et al and also by chromatofocussing. The final preparation appeared homogenous by SDS-PAGE with an apparent molecular weight of 52,000, had a specific content of 11 nmoles P-450 per mg protein, showed very high activity of low K/sub m/ dimethylnitrosamine demethylase and produced a type II binding spectrum with dimethylsulfoxide. The enzyme was also active with aniline and aminopyrine as substrates. Pyrazole itself served as an excellent substrate with 4-hydroxy pyrazole being the product. An antibody against the pyrazole-inducible P-450 raised in chickens recognized a protein with mol.wt of about 52,000 in control microsomes. This band was highly enriched in microsomes from rats treated with pyrazole, 4-methyl-pyrazole, ethanol or acetone, but not phenobarbital or 3-methylcholanthrene. In summary, the pyrazole-inducible P-450 has been purified and appears to be identical in its catalytic and immunological properties to the alcohol-inducible P-450.},

  doi          = {},

  url          = {https://www.osti.gov/biblio/6290676},
  journal      = {Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)},
number       = ,

  volume       = 46:6,

  place        = {United States},

  year         = {Fri May 01 00:00:00 EDT 1987},

  month        = {Fri May 01 00:00:00 EDT 1987}

}

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