title: N-oxygenation of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine by the rat liver flavin-containing monooxygenase expressed in yeast cells.
abstract: N-oxygenation of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), a dopaminergic neurotoxin, was studied using recombinant rat liver flavin-containing monooxygenase (FMO), FMO1A1, expressed in yeast cells. The mean (+/- S.D.) kinetic parameters of MPTP N-oxygenation were: Km = 1.8 +/- 0.5 microM, Vmax = 9.5 +/- 1.6 nmol/mg per min, and Vmax/Km = 4.6 +/- 0.5 ml/mg per min. n-Octylamine, an activator of FMO, enhanced the MPTP N-oxygenation activity by 51%, while methimazole, thiobenzamide and alpha-naphthylthiourea, alternate substrates of FMO, inhibited it by 27.4, 68.0 and 59.2%, respectively. The results indicate that MPTP is efficiently N-oxygenated by the recombinant rat liver FMO1A1, and that the responses to the modulators of FMO activity found in the recombinant rat liver FMO1A1 resemble those of mouse and rat liver microsomes as reported previously. The findings suggest that the recombinant FMO expressed in yeast cells is considered as a useful tool to study an involvement of FMO in the metabolism of environmental toxins or chemicals. In addition, FMO1A1 appears to be one of the predominant enzymes responsible for the N-oxygenation of MPTP at least in rat liver.
authors.affiliation: Department of Clinical Pharmacology, International Medical Center of Japan, Tokyo.
authors.forename: K
authors.initials: K
authors.lastname: Chiba
authors.forename: K
authors.initials: K
authors.lastname: Kobayashi
authors.forename: K
authors.initials: K
authors.lastname: Itoh
authors.forename: S
authors.initials: S
authors.lastname: Itoh
authors.forename: T
authors.initials: T
authors.lastname: Chiba
authors.forename: T
authors.initials: T
authors.lastname: Ishizaki
authors.forename: T
authors.initials: T
authors.lastname: Kamataki
mesh_terms: D015632:1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine / Q000378:metabolism*; D000818:Animals; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D010105:Oxygenases / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology*
chemical_list: D011994:Recombinant Proteins; D015632:1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine; D010105:Oxygenases; C020066:dimethylaniline monooxygenase (N-oxide forming)