Purification of a cytochrome P450 isozyme belonging to a subfamily of P450 IIB from liver microsomes of guinea pigs.
An isozyme of cytochrome P450 was purified from liver microsomes of guinea pigs by HPLC with anion-exchange and hydroxylapatite columns. The isozyme showed a single band of 52 kdalton on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Oxidation activities of the final preparation towards p-nitroanisole, aniline and d-benzphetamine were 1.3 to 15.4-fold comparing with those of microsomal fraction. This isozyme was also concerned with oxidation of delta 9-tetrahydrocannabinol (THC) to 8 alpha-hydroxy- (8 alpha-OH-), 2'-OH- and 3'-OH-delta 9-THCs. The N-terminal region of the isozyme was considerably hydrophobic, and 13 of the first 20 amino acid residues was leucine. Since this first 20 amino acid sequence is 80% homologous with that of cytochrome P450 LM2 purified from rabbit liver microsomes, this isozyme can be categorized to a subfamily of P450 IIB.
Narimatsu
S
S
Department of Hygienic Chemistry, Faculty of Pharmaceutical Sciences, Hokuriku University, Kanazawa, Japan.
Akutsu
Y
Y
Matsunaga
T
T
Watanabe
K
K
Yamamoto
I
I
Yoshimura
H
H
D000595:Amino Acid Sequence; D000818:Animals; D002845:Chromatography; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D003577:Cytochrome P-450 Enzyme System / Q000302:isolation & purification* / Q000378:metabolism; D017886:Durapatite; D004591:Electrophoresis, Polyacrylamide Gel; D006168:Guinea Pigs; D006882:Hydroxyapatites; D007527:Isoenzymes / Q000302:isolation & purification* / Q000378:metabolism; D007700:Kinetics; D008297:Male; D008862:Microsomes, Liver / Q000201:enzymology*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D012689:Sequence Homology, Nucleic Acid; D013379:Substrate Specificity
D006882:Hydroxyapatites; D007527:Isoenzymes; D003577:Cytochrome P-450 Enzyme System; D017886:Durapatite