[Isolation and characteristics of coumarin-specific cytochrome P-450 (P-450Cho) from liver microsomes of DBA/2N mice, induced with pyrazole].
Coumarin-specific cytochrome P-450 (P-450Coh) has been isolated from liver microsomes of DBA/2N mice induced with pyrazole. The induction effect was accompanied by a 5.8-5.9-fold increase in the P-450Coh content which made up to 14.4-17% of the total cytochrome P-450 pool in the microsomes. At the final step of P-450Coh purification, variously substituted Sepharoses (hydroxyphenyl-, cholate-, aminooctyl- and t-cytochrome-b5-) were used. The optimal scheme involved solubilization of microsomes with sodium cholate, hydrophobic chromatography on octyl-Sepharose, adsorption on calcium-tartrate gel and hydrophobic ion-exchange chromatography on aminooctyl-Sepharose. According to SDS gel electrophoresis data, the purity of P-450Coh was 95% and Mr was 50,000 Da. The amino acid composition of the protein includes 445 residues. At saturating concentrations of coumarin, more than 90% of P-450Coh are represented by the high spin form. The catalytic activity of P-450Coh was studied in reactions of xenobiotics oxidation.
Shkumatov
V M
VM
Chashchin
V L
VL
Akhrem
A A
AA
Iuvonen
R O
RO
Lang
M A
MA
D000818:Animals; D003374:Coumarins / Q000378:metabolism*; D003580:Cytochromes / Q000096:biosynthesis / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D004790:Enzyme Induction; D006900:Hydroxylation; D051379:Mice; D008811:Mice, Inbred DBA; D008862:Microsomes, Liver / Q000201:enzymology*; D010634:Phenobarbital / Q000494:pharmacology; D011720:Pyrazoles / Q000494:pharmacology*; D012995:Solubility; D013056:Spectrophotometry, Ultraviolet; D013379:Substrate Specificity
D003374:Coumarins; D003580:Cytochromes; D011720:Pyrazoles; C030123:coumarin; D010634:Phenobarbital
Vydelenie i kharakteristika kumarinspetsifichnogo tsitokhroma P-450 (P-450Coh) iz mikrosom pecheni mysheĭ DBA/2N, indutsirovannykh pirazolom.