Cloned (Comment) | Organism |
---|---|
expression of wild-type and mtant enzymes in Escherichia coli strain JM109 | Homo sapiens |
Crystallization (Comment) | Organism |
---|---|
purified recombinant mutant E308V, 15mg/ml protein in 5 mM Tris, pH 8.5, containing 1mM sodium salicylate and 0.1mM EDTA, incubation with 5 mM dithiothreitol for 1 h at room temperature, aliquots of 0.005 ml protein solution are mixed with 0.005 ml of 100 mM sodium citrate, pH 5.0, containing 8-11% polyethylene glycol 8000, 5 mM DTT, 1 mM sodium salicylate, 0.1 mM EDTA, and placed on siliconized glass plates, 7 days at 20°C, X-ray diffraction structure determination and analysis at 2.6 A resolution | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
E308V | site-directed mutagenesis, mutant molybdopterin domain structure, overview, the xanthine oxidase changes its substrate specificity to aldehyde oxidase type upon mutation of amino acid residues in the active site, the E803V mutation almost completely abrogates the activity towards hypoxanthine as a substrate, but the decrease in activity towards purine substrate is not due to large conformational change in the mutant enzyme | Homo sapiens |
R881M | site-directed mutagenesis, the xanthine oxidase changes its substrate specificity to aldehyde oxidase type upon mutation of amino acid residues in the active site, the R881M mutant lacks activity towards xanthine, but retaines slight activity towards hypoxanthine | Homo sapiens |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
allopurinol | - |
Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | steady-state kinetics analysis of wild-type and mutant enzymes with hypoxanthine of xanthine, and of mutant enzymes with benzaldehyde or 4-(dimethyamino)cinnamaldehyde as aldehyde oxidase substrates, overview | Homo sapiens | |
0.0017 | - |
hypoxanthine | pH 8.5, 25°C, recombinant wild-type enzyme | Homo sapiens | |
0.0088 | - |
xanthine | pH 8.5, 25°C, recombinant wild-type enzyme | Homo sapiens | |
0.021 | - |
hypoxanthine | pH 8.5, 25°C, recombinant mutant R881M | Homo sapiens | |
0.072 | - |
xanthine | pH 8.5, 25°C, recombinant mutant E308V | Homo sapiens |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Molybdenum | molybdopterin domain structure of enzyme mutant E308V, overview | Homo sapiens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
hypoxanthine + 2 H2O + 2 O2 | Homo sapiens | - |
urate + 2 H2O2 | - |
? | |
xanthine + H2O + O2 | Homo sapiens | roles of active site residues E308 and R881 in binding and activation of purine substrate, overview | urate + H2O2 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Escherichia coli strain JM109 by anion exchange and hydroxyapatite chromatography, ultrafiltration, and gel filtration | Homo sapiens |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
xanthine + H2O + O2 = urate + H2O2 | hydrogen-bonding arrangement of the substrate-bound complex at the active site, and proposed mechanism of the purine oxidation reaction by XOR | Homo sapiens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
hypoxanthine + 2 H2O + 2 O2 | - |
Homo sapiens | urate + 2 H2O2 | - |
? | |
xanthine + H2O + O2 | - |
Homo sapiens | urate + H2O2 | - |
? | |
xanthine + H2O + O2 | roles of active site residues E308 and R881 in binding and activation of purine substrate, overview | Homo sapiens | urate + H2O2 | - |
? |
Synonyms | Comment | Organism |
---|---|---|
xanthine oxidoreductase | - |
Homo sapiens |
XOR | - |
Homo sapiens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Homo sapiens |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.33 | - |
hypoxanthine | pH 8.5, 25°C, recombinant mutant R881M | Homo sapiens | |
1.35 | - |
xanthine | pH 8.5, 25°C, recombinant mutant E308V | Homo sapiens | |
18.3 | - |
xanthine | pH 8.5, 25°C, recombinant wild-type enzyme | Homo sapiens | |
30 | - |
hypoxanthine | pH 8.5, 25°C, recombinant wild-type enzyme | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.5 | - |
assay at | Homo sapiens |