title: Characteristic properties and kinetic analysis with neurotoxins of porcine FAD-containing monooxygenase.
abstract: An FAD-containing monooxygenase (EC 1.14.13.8) was purified from porcine liver microsomes by a new purification procedure and confirmed to give an electrophoretically single protein band. The optical and CD spectra, fluorescence and molar extinction coefficients of the FMO were investigated. The activity of the FMO was examined kinetically with neurotoxins, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), 1,2,3,4-tetrahydroisoquinoline (TIQ), and 1-methyl-6,7-dihydroxytetrahydroisoquinoline (MDTIQ), as substrates. The kinetic parameters of the FMO for the neurotoxins, molecular oxygen and electron donors were determined, in comparison with those of dimethylaniline. The CD spectrum of the FMO was measured in the absence and presence of NADP+, dimethylaniline or both. The results showed that the FMO metabolized the neurotoxins, and that NADH was a weak electron donor for it. The CD spectrum of the FMO in the oxidized form, which acts as an oxidase and oxygenase, unlike that of D-amino-acid oxidase, showed negative ellipticity, the secondary structure of the FMO changed, and the alpha-helix structure of the monooxygenase was affected by the formation of a complex of the FMO with NADP+, DMA or both.
authors.affiliation: Department of Biochemistry, Kagawa Medical School, Japan.
authors.forename: R F
authors.initials: RF
authors.lastname: Wu
authors.forename: Y
authors.initials: Y
authors.lastname: Ichikawa
mesh_terms: D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002942:Circular Dichroism; D007700:Kinetics; D008862:Microsomes, Liver / Q000201:enzymology; D015394:Molecular Structure; D009498:Neurotoxins / Q000378:metabolism*; D010100:Oxygen / Q000378:metabolism; D010105:Oxygenases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D013552:Swine
chemical_list: D000596:Amino Acids; D009498:Neurotoxins; D010105:Oxygenases; C020066:dimethylaniline monooxygenase (N-oxide forming); D010100:Oxygen