Purification and properties of purine hydroxylase II from Aspergillus nidulans.
Purine hydroxylase II from Aspergillus nidulans has been purified to near homogeneity. The enzyme has a pI of 5.7, a molecular weight of 300,000, and two subunits with molecular weight of 153,000 each. The enzyme contains 2 FAD, 2 molybdenum atoms, and 4 (2 Fe-2S) iron-sulfur centers per molecule and exhibits broad specificity for reducing and oxidizing substrates. Among the more notable characteristics are the ability to oxidize hypoxanthine and nicotinic acid but not xanthine and virtually complete inactivity with oxygen. Moreover, while the enzyme is inactivated by borate and methanol, it is very resistant to cyanide and arsenite and it not inactivated by allopurinol. At infinite concentrations of reducing and oxidizing substrates, the Km for hypoxanthine was 119 microM, for nicotinic acid was 136 microM, and for NAD+ was 525 microM.
D000429:Alcohol Oxidoreductases / Q000302:isolation & purification*; D001233:Aspergillus nidulans / Q000201:enzymology*; D005182:Flavin-Adenine Dinucleotide / Q000032:analysis; D006863:Hydrogen-Ion Concentration; D007506:Iron-Sulfur Proteins / Q000302:isolation & purification; D007700:Kinetics; D046911:Macromolecular Substances; D008970:Molecular Weight; D008982:Molybdenum / Q000032:analysis; D013379:Substrate Specificity
D007506:Iron-Sulfur Proteins; D046911:Macromolecular Substances; D005182:Flavin-Adenine Dinucleotide; D008982:Molybdenum; D000429:Alcohol Oxidoreductases; C040748:purine hydroxylase II