A rabbit liver constitutive form of cytochrome P450 responsible for amphetamine deamination.
A cytochrome P450 isozyme responsible for amphetamine deamination was purified from hepatic microsomes of untreated rabbits. The purification procedures consisted of a set of column chromatographies with omega-aminooctyl-Sepharose 4B, DEAE-cellulose, CM-Sephadex C-50, and hydroxyapatite. The deamination activity was determined by measuring the formation of phenylacetone after derivatization to the p-nitrobenzyloxim by HPLC. This isozyme, which was designated P450APD, showed a monomeric molecular weight of 51,000 in sodium dodecyl sulfate-polyacrylamide gel electrophoresis and exhibited an absorption maximum of reduced CO complex at 451 nm. On the basis of the specificity toward testosterone metabolism and the N-terminal amino acid sequence, P450APD was attributed to a member of P450 class IIC subfamily, which is identical or closely related to LM3b (D. R. Koop and M. J. Coon (1979) Biochem, Biophys, Res. Commun. 91, 1075-1081), form 3b (E. F. Johnson (1980) J. Biol. Chem. 255, 304-309), and other similar preparations. Antibody against the P450APD inhibited about 80% of the amphetamine deamination activity in rabbit hepatic microsomes as well as in the reconstitution system of this P450. The present results support that P450APD is the major P450 isozyme responsible for amphetamine deamination in rabbit liver.
Yamada
H
H
Faculty of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan.
Honda
S
S
Oguri
K
K
Yoshimura
H
H
D000661:Amphetamine / Q000378:metabolism*; D000818:Animals; D002848:Chromatography, DEAE-Cellulose; D002852:Chromatography, Ion Exchange; D003577:Cytochrome P-450 Enzyme System / Q000302:isolation & purification / Q000378:metabolism*; D003641:Deamination; D004591:Electrophoresis, Polyacrylamide Gel; D007527:Isoenzymes / Q000302:isolation & purification / Q000378:metabolism*; D008297:Male; D008862:Microsomes, Liver / Q000201:enzymology*; D011817:Rabbits
D007527:Isoenzymes; D003577:Cytochrome P-450 Enzyme System; D000661:Amphetamine