Characteristic properties and kinetic analysis with neurotoxins of porcine FAD-containing monooxygenase

Ru Feng Wu, Yoshiyuki Ichikawa

Research output: Contribution to journalArticle

8 Citations (Scopus)

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-l,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, on 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 α-helix structure of the monooxygenase was affected by the formation of a complex of the FMO with NADP+, DMA or both.

Original languageEnglish (US)
Pages (from-to)204-210
Number of pages7
JournalBiochimica et Biophysica Acta (BBA)/Protein Structure and Molecular
Volume1208
Issue number2
DOIs
StatePublished - Oct 19 1994

Fingerprint

dimethylaniline monooxygenase (N-oxide forming)
Flavin-Adenine Dinucleotide
Neurotoxins
Mixed Function Oxygenases
Swine
NADP
Kinetics
Oxidoreductases
Electrons
Oxygenases
Molecular oxygen
Liver Microsomes
Dynamic mechanical analysis
Kinetic parameters
Liver
NAD
Purification
Fluorescence
Oxygen
Amino Acids

Keywords

  • FAD-containing monooxygenase
  • Flavoenzyme
  • Kinetics
  • Neurotoxin
  • Oxygenase

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology
  • Structural Biology

Cite this

Characteristic properties and kinetic analysis with neurotoxins of porcine FAD-containing monooxygenase. / Wu, Ru Feng; Ichikawa, Yoshiyuki.

In: Biochimica et Biophysica Acta (BBA)/Protein Structure and Molecular, Vol. 1208, No. 2, 19.10.1994, p. 204-210.

Research output: Contribution to journalArticle

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AB - 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-l,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, on 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 α-helix structure of the monooxygenase was affected by the formation of a complex of the FMO with NADP+, DMA or both.

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