Purification of cytochromes P-448 from β-naphthoflavone-treated rainbow trout |
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Authors: | David E Williams Donald R Buhler |
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Institution: | Department of Agricultural Chemistry and Environmental Health Sciences Center, Oregon State University, Corvallis, OR 97331, U.S.A. |
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Abstract: | Rainbow trout were treated with β-naphthoflavone and the hepatic microsomal cytochrome P-450 solubilized with 3-(3-cholaminopropyl)dimethylammonio]-1-propanesulfonate. Chromatography on tryptamine-Sepharose 4B gave a single cytochrome P-450 peak which was further resolved into three components by elution from DEAE-Sepharose. The two main peaks were then chromatographed on hydroxyapatite and a total of four fractions obtained. Two of these fractions had similar properties and significantly metabolized in a reconstituted system containing rat cytochrome P-450 reductase. This activity was inhibited by α-naphthoflavone but not by metyrapone or SKF-525A. Purified cytochromes P-448 from 3-methylcholanthrene-treated rat had similar spectral properties and activity towards suggesting similarities between these forms. |
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Keywords: | Cytochrome P-448 Naphthoflavone (Rainbow trout) CHAPS 3-[(3-cholamidopropyl)dimethylamonio]- 1-propanesulfonate EDC 1-ethyl-3-(3-dimethyl- aminopropyl)carbodiimide hydrochloride |
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