Cloning and functional expression of human kynurenine 3-monooxygenase.
Kynurenine 3-monooxygenase, an NADPH-dependent flavin monooxygenase, catalyses the hydroxylation of L-kynurenine to L-3-hydroxykynurenine. By hybridization screening using a cDNA probe encoding the entire exon 2 of Drosophila melanogaster kynurenine 3-monooxygenase, we isolated a 2.0 kb cDNA clone coding for the corresponding human liver enzyme. The deduced amino acid sequence of the human protein consists of 486 amino acids with a predicted molecular mass of 55,762 Da. Transfection of the human cDNA in HEK-293 cells resulted in the functional expression of the enzyme with kinetic properties similar to those found for the native human protein. RNA blot analysis of human tissues revealed the presence of a major mRNA species of approximately 2.0 kb in liver, placenta and kidney.
Alberati-Giani
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Pharmaceutical Division, Preclinical Research, F. Hoffmann-La Roche Ltd, Basel, Switzerland. daniela.alberati@roche.com
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Broger
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Warren
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Röver
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Malherbe
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D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004331:Drosophila melanogaster / Q000201:enzymology; D015870:Gene Expression; D006801:Humans; D050601:Kynurenine 3-Monooxygenase; D008099:Liver / Q000201:enzymology*; D006899:Mixed Function Oxygenases / Q000235:genetics*; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000235:genetics; D017421:Sequence Analysis; D017386:Sequence Homology, Amino Acid
D018076:DNA, Complementary; D011993:Recombinant Fusion Proteins; D006899:Mixed Function Oxygenases; D050601:Kynurenine 3-Monooxygenase