Regulation and biochemistry of mouse molybdo-flavoenzymes. The DBA/2 mouse is selectively deficient in the expression of aldehyde oxidase homologues 1 and 2 and represents a unique source for the purification and characterization of aldehyde oxidase.
Mouse molybdo-flavoenzymes consist of xanthine oxidoreductase, aldehyde oxidase (AOX1), and two recently identified proteins, AOH1 and AOH2 (aldehyde oxidase homologues 1 and 2). Here we demonstrate that CD-1, C57BL/6, 129/Sv, and other mouse strains synthesize high levels of AOH1 in the liver and AOH2 in the skin. By contrast, the DBA/2 and CBA strains are unique, having a selective deficit in the expression of the AOH1 and AOH2 genes. DBA/2 animals synthesize trace amounts of a catalytically active AOH1 protein. However, relative to CD-1 animals, an over 2 log reduction in the steady-state levels of liver AOH1 mRNA, protein, and enzymatic activity is observed in basal conditions and following administration of testosterone. The DBA/2 mouse represents a unique opportunity to purify AOX1 and compare its enzymatic characteristics to those of the AOH1 protein. The spectroscopy and biochemistry of AOX1 are very similar to those of AOH1 except for a differential sensitivity to the non-competitive inhibitory effect of norharmane. AOX1 and AOH1 oxidize an overlapping set of aldehydes and heterocycles. For most compounds, the substrate efficiency (V(max)/K(m)) of AOX1 is superior to that of AOH1. Alkylic alcohols and acetaldehyde, the toxic metabolite of ethanol, are poor substrates of both enzymes. Consistent with this, the levels of acetaldehyde in the livers of ethanol administered CD-1 and DBA/2 mice are similar, indicating that neither enzyme is involved in the in vivo biotransformation of acetaldehyde.
Vila
Ruth
R
Department of Biochemistry and Molecular Pharmacology, Istituto di Ricerche Farmacologiche, "Mario Negri," via Eritrea, 62, Milano 20157, Italy.
Kurosaki
Mami
M
Barzago
Maria Monica
MM
Kolek
Metodej
M
Bastone
Antonio
A
Colombo
Laura
L
Salmona
Mario
M
Terao
Mineko
M
Garattini
Enrico
E
D000445:Aldehyde Oxidoreductases / Q000172:deficiency* / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D000431:Ethanol / Q000008:administration & dosage; D005420:Flavoproteins / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic; D051379:Mice; D008811:Mice, Inbred DBA / Q000235:genetics / Q000378:metabolism*; D008815:Mice, Inbred Strains; D008969:Molecular Sequence Data; D009928:Organ Specificity; D013045:Species Specificity
D005420:Flavoproteins; D000431:Ethanol; D000445:Aldehyde Oxidoreductases; C443116:Aox3 protein, mouse; C443117:aldehyde oxidase homolog 2, mouse