Initial purification and characterization of hepatic microsomal cytochrome P-450 from BNF-treated perch (Perca fluviatilis).
1. A procedure was developed for isolating and purifying cytochrome P-450 from hepatic microsomes of BNF-treated perch, using modified versions of the methods of Williams and Buhler (1982. Biochim. biophys. Acta 717, 398-404) and Goksøyr (1985. Biochim. biophys. Acta 850, 409-417). 2. Following chromatography on phenyl-Sepharose CL 4B and DEAE-Sepharose CL-6B, the major peaks, fractions b and c, were resolved into five fractions, possibly representing different isoenzymes, by a FPLC with a strong anion exchange column (Mono Q). 3. These fractions have been characterized on the basis of their spectral, electrophoretic and immunological properties. 4. The purified form of cytochrome P-450 in fraction V from perch liver showed a number of similarities to cytochrome P-450c, the major BNF-inducible cytochrome P-450 in cod liver. 5. Therefore we suggest that this purified form of cytochrome P-450 is a BNF-induced form in perch and that it is closely related to the gene subfamily cytochrome P-450 IA1.
Zhang
Y S
YS
Department of Zoophysiology, University of Göteborg, Sweden.
Goksøyr
A
A
Andersson
T
T
Förlin
L
L
D000818:Animals; D001571:Benzoflavones / Q000494:pharmacology*; D002851:Chromatography, High Pressure Liquid; D019363:Cytochrome P-450 CYP1A1; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D004790:Enzyme Induction / Q000187:drug effects; D015151:Immunoblotting; D008862:Microsomes, Liver / Q000187:drug effects / Q000201:enzymology*; D008970:Molecular Weight; D010088:Oxidoreductases / Q000096:biosynthesis; D010471:Perches / Q000378:metabolism*; D013053:Spectrophotometry; D019324:beta-Naphthoflavone
D001571:Benzoflavones; D019324:beta-Naphthoflavone; D003577:Cytochrome P-450 Enzyme System; D010088:Oxidoreductases; D019363:Cytochrome P-450 CYP1A1