Cloning and functional expression of human kynurenine 3-monooxygenase

• Published in: FEBS letters Vol. 410; no. 2; pp. 407 - 412
• Main Authors: Alberati-Giani, Daniela, Cesura, Andrea M, Broger, Clemens, Warren, William D, Röver, Stephan, Malherbe, Pari
• Format: Journal Article
• Language: English
• Published: England Elsevier B.V 30.06.1997
• Subjects: Amino Acid Sequence; Animals; base pair(s); Base Sequence; Blotting, Northern; Cloning, Molecular; dmKMO; DNA, Complementary; Drosophila melanogaster; Drosophila melanogaster - enzymology; Drosophila melanogaster kynurenine 3-monooxygenase; Flavin monooxygenase; Gene Expression; hKMO; human kynurenine 3-monooxygenase; Human liver; Humans; kilobases; KMO; Kynurenine 3-Monooxygenase; Kynurenine pathway; Liver - enzymology; Mixed Function Oxygenases - genetics; Molecular Sequence Data; open reading frame; ORF; Recombinant Fusion Proteins - genetics; Sequence Analysis; Sequence comparison; Sequence Homology, Amino Acid; ORF; hKMO; Human liver; dmKMO; kb; Kynurenine 3-monooxygenase; Flavin monooxygenase; Sequence comparison; KMO; Drosophila melanogaster; bp; Kynurenine pathway
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/S0014-5793(97)00627-3

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 ∼2.0 kb in liver, placenta and kidney.