Functional activity of the mouse flavin-containing monooxygenase forms 1, 3, and 5
| Autor: | John R. Cashman, Ruzbeh Mosadeghi, Jun Zhang, Matt Lawson, Matt A. Cerny |
|---|---|
| Rok vydání: | 2007 |
| Předmět: |
Male
Hot Temperature Recombinant Fusion Proteins Health Toxicology and Mutagenesis Flavin-containing monooxygenase Toxicology medicine.disease_cause Biochemistry Chromatography Affinity Maltose-Binding Proteins Substrate Specificity law.invention Mice Structure-Activity Relationship Maltose-binding protein Affinity chromatography law Enzyme Stability Escherichia coli medicine Animals Cloning Molecular Molecular Biology chemistry.chemical_classification Molecular Structure biology General Medicine Hydrogen-Ion Concentration Monooxygenase Fusion protein Isoenzymes Mice Inbred C57BL Kinetics Enzyme Liver chemistry Oxygenases Recombinant DNA biology.protein Molecular Medicine Carrier Proteins NADP |
| Zdroj: | Journal of Biochemical and Molecular Toxicology. 21:206-215 |
| ISSN: | 1099-0461 1095-6670 |
| DOI: | 10.1002/jbt.20176 |
| Popis: | Three functional mouse flavin-containing monooxygenases (mFMOs) (i.e., mFMO1, mFMO3, and mFMO5) have been reported to be the major FMOs present in mouse liver. To examine the biochemical features of these enzymes, recombinant enzymes were expressed as maltose-binding protein fusion proteins (i.e., MBP-mFMO1, MBP-mFMO3, and MBP-mFMO5) in Escherichia coli and isolated and purified with affinity chromatography. The substrate specificity of these three mouse hepatic FMO enzymes were examined using a variety of substrates, including mercaptoimidazole, trimethylamine, S-methyl esonarimod, and an analog thereof, and a series of 10-(N,N-dimethylaminoalkyl)-2-(trifluoromethyl)phenothiazine analogs. The kinetic parameters of the three mouse FMOs for these substrates were compared in an attempt to explore substrate structure--function relationships specific for each mFMO. Utilizing a common phenothiazine substrate for all three enzymes, we compared the pH dependence for the recombinant enzymes under similar conditions. In addition, thermal stability for mFMO1, mFMO3, and mFMO5 enzymes was examined in the presence and absence of NADPH. The results revealed unique features for mFMO5, suggesting possible impact on the functional significance of this abundantly expressed FMO5 isoform in both human and mouse liver. |
| Databáze: | OpenAIRE |
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