Purification of the aldehyde oxidase homolog 1 (AOH1) protein and cloning of the AOH1 and aldehyde oxidase homolog 2 (AOH2) genes - Identification ofa novel molybdo-flavoprotein gene cluster on mouse chromosome 1
M. Terao et al., Purification of the aldehyde oxidase homolog 1 (AOH1) protein and cloning of the AOH1 and aldehyde oxidase homolog 2 (AOH2) genes - Identification ofa novel molybdo-flavoprotein gene cluster on mouse chromosome 1, J BIOL CHEM, 276(49), 2001, pp. 46347-46363
We report the cloning of the AOH1 and AOH2 genes, which encode two novel ma
mmalian molybdo-flavoproteins. We have purified the AOH1 protein to homogen
eity in its catalytically active form from mouse liver. Twenty tryptic pept
ides, identified or directly sequenced by mass spectrometry, confirm the pr
imary structure of the polypeptide deduced from the AOH1 gene. The enzyme c
ontains one molecule of FAD, one atom of molybdenum, and four atoms of iron
per subunit and shows spectroscopic features similar to those of the proto
typic molybdo-flavoprotein xanthine oxidoreductase. The AOH1 and AOH2 genes
are 98 and 60 kilobases long, respectively, and consist of 35 coding exons
. The AOH1 gene has the potential to transcribe an extra leader non-coding
exon, which is located downstream of exon 26, and is transcribed in the opp
osite orientation relative to all the other exons. AOH1 and AOH2 map to chr
omosome I in close proximity to each other and to the aldehyde oxidase gene
, forming a molybdo-flavoenzyme gene cluster. Conservation in the position
of exon/intron junctions among the mouse AOH1, AOH2, aldehyde oxidase, and
xanthine oxidoreductase loci indicates that these genes are derived from th
e duplication of an ancestral precursor.