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Literature summary for 1.7.3.3 extracted from
- Structure-based characterization of canine-human chimeric uricases and its evolutionary implications (2012), Biochimie, 94, 1412-1420.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expression in Escherichia coli | Canis lupus familiaris |
Crystallization (Commentary)
| Crystallization (Comment) | Organism |
|---|---|
| homology modeling of monomeric enzyme. The highly conserved residue Gly290 could interact with Asn262 and His264. Residue substitutions near Gly290 may affect its spatial orientation and result in changes in catalysis.Gly290 is likely to participate in the structure of the active site and to be involved in oxygen-binding | Canis lupus familiaris |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| A296V | about 50% ioncrease in catalytic efficiency | Canis lupus familiaris |
| A89T/G91A7V92M/H245L/E252A/M253I/R291K/A296V/A301S/K303R | replacement with the corresponding residues of human enzyme. About 30% of wild-type catalytic efficiency | Canis lupus familiaris |
| H245L/E252A/M253I/R291K/A296V/A301S/K303R | replacement with the corresponding residues of human enzyme. Catalytic efficiency is higher than in wild-type, but below the efficiency of mutant R291K/A296V/A301S/K303R | Canis lupus familiaris |
| I115V/H119R/L120F/H245L/E252A/M253I/R291K/A296V/A301S/K303R | replacement with the corresponding residues of human enzyme, complete loss of activity | Canis lupus familiaris |
| R119H | complete loss of activity | Canis lupus familiaris |
| R291K/A296V/A301S/K303R | replacement with the corresponding residues of human enzyme. About 50% increase in catalytic efficiency | Canis lupus familiaris |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.0064 | - |
Urate | mutant A296V, pH 8.6, 25°C | Canis lupus familiaris |  |
| 0.0064 | - |
Urate | mutant R291K/A296V/A301S/K303R, pH 8.6, 25°C | Canis lupus familiaris | |
| 0.0065 | - |
Urate | mutant H245L/E252A/M253I/R291K/A296V/A301S/K303R, pH 8.6, 25°C | Canis lupus familiaris | |
| 0.0085 | - |
Urate | wild-type, pH 8.6, 25°C | Canis lupus familiaris | |
| 0.017 | - |
Urate | mutant A89T/G91A7V92M/H245L/E252A/M253I/R291K/A296V/A301S/K303R, pH 8.6, 25°C | Canis lupus familiaris | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Canis lupus familiaris | Q5FZI9 | - |
- |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 2.99 | - |
mutant A89T/G91A7V92M/H245L/E252A/M253I/R291K/A296V/A301S/K303R, pH 8.6, 25°C | Canis lupus familiaris |
| 4.22 | - |
wild-type, pH 8.6, 25°C | Canis lupus familiaris |
| 4.91 | - |
mutant H245L/E252A/M253I/R291K/A296V/A301S/K303R, pH 8.6, 25°C | Canis lupus familiaris |
| 5.68 | - |
mutant A296V, pH 8.6, 25°C | Canis lupus familiaris |
| 5.94 | - |
mutant R291K/A296V/A301S/K303R, pH 8.6, 25°C | Canis lupus familiaris |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| urate + O2 + H2O | - |
Canis lupus familiaris | 5-hydroxyisourate + H2O2 | - |
? | |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 8.7 | - |
Urate | mutant A89T/G91A7V92M/H245L/E252A/M253I/R291K/A296V/A301S/K303R, pH 8.6, 25°C | Canis lupus familiaris | |
| 13.6 | - |
Urate | wild-type, pH 8.6, 25°C | Canis lupus familiaris | |
| 15.5 | - |
Urate | mutant H245L/E252A/M253I/R291K/A296V/A301S/K303R, pH 8.6, 25°C | Canis lupus familiaris | |
| 16.1 | - |
Urate | mutant A296V, pH 8.6, 25°C | Canis lupus familiaris | |
| 16.8 | - |
Urate | mutant R291K/A296V/A301S/K303R, pH 8.6, 25°C | Canis lupus familiaris | |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 510 | - |
Urate | mutant A89T/G91A7V92M/H245L/E252A/M253I/R291K/A296V/A301S/K303R, pH 8.6, 25°C | Canis lupus familiaris | |
| 1580 | - |
Urate | wild-type, pH 8.6, 25°C | Canis lupus familiaris | |
| 2390 | - |
Urate | mutant H245L/E252A/M253I/R291K/A296V/A301S/K303R, pH 8.6, 25°C | Canis lupus familiaris | |
| 2510 | - |
Urate | mutant A296V, pH 8.6, 25°C | Canis lupus familiaris | |
| 2620 | - |
Urate | mutant R291K/A296V/A301S/K303R, pH 8.6, 25°C | Canis lupus familiaris | |
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