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Literature summary for 3.5.2.10 extracted from
- Substitution of Glu122 by glutamine revealed the function of the second water molecule as a proton donor in the binuclear metal enzyme creatininase (2010), J. Mol. Biol., 396, 1081-1096.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
- |
Pseudomonas putida |
Crystallization (Commentary)
| Crystallization (Comment) | Organism |
|---|---|
| crystal structure of the enzyme complexed with a 1-methylguanidine. All subunits in the complex exist as the closed form. X-ray crystallographic study and the atomic absorption spectrometry analysis of the E122Q mutant-substrate complex reveal that the drastic decrease of the activity of the E122Q is caused by the loss of one Zn ion at the Metal1 site and a critical function of Glu122 | Pseudomonas putida |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| E122Q | almost complete loss of activity | Pseudomonas putida |
| E183Q | complete loss of activity | Pseudomonas putida |
| H178A | complete loss of activity | Pseudomonas putida |
| W154A | complete loss of activity | Pseudomonas putida |
| W154F | almost complete loss of activity | Pseudomonas putida |
| W174A | almost complete loss of activity | Pseudomonas putida |
| W174F | about 50% of wild-type activity | Pseudomonas putida |
| Y121A | about 10% of wild-type catalytic efficiency | Pseudomonas putida |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| EDTA | wild-type enzyme, purified in the presence of EDTA, shows only half the activity of the wild-type enzyme, the activity is recovered by the addition of ZnCl2 or MnCl2 | Pseudomonas putida |  |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 44 | - |
creatinine | wild-type, presence of Zn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida | |
| 56 | - |
creatinine | mutant E122Q, presence of Mn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida | |
| 77 | - |
creatinine | wild-type, presence of Mn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida | |
| 90 | - |
creatinine | mutant W174F, presence of Mn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida | |
| 150 | - |
creatinine | wild-type, presence of EDTA, semi-apo-enzyme, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida | |
| 160 | - |
creatinine | mutant Y121A, presence of Mn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida | |
| 220 | - |
creatinine | mutant E122Q, presence of Zn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida | |
| 350 | - |
creatinine | mutant W154F, presence of Mn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Zinc | removal of the zinc ions by dialyzing against buffers containing o-phenanthroline results in a partial decrease of 10-30% in activity and zinc content. Wild-type enzyme, purified in the presence of EDTA, shows only half the activity of the wild-type enzyme, the activity is recovered by the addition of ZnCl2 or MnCl2 | Pseudomonas putida | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Pseudomonas putida | P83772 | - |
- |
Reaction
| Reaction | Comment | Organism | Reaction ID |
|---|---|---|---|
| creatinine + H2O = creatine | catalytic mechanism: a first water molecule is a hydroxide ion that is bound as a bridge between the two metal ions and attacks the carbonyl carbon of the substrate. A second water molecule is bound to the carboxyl group of Glu122 and functions as a proton donor in catalysis | Pseudomonas putida |
Renatured (Commentary)
| Renatured (Comment) | Organism |
|---|---|
| wild-type enzyme, purified in the presence of EDTA, shows only half the activity of the wild-type enzyme, the activity is recovered by the addition of ZnCl2 or MnCl2 | Pseudomonas putida |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 0.13 | - |
mutant E122Q, presence of Zn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida |
| 4.9 | - |
mutant W154F, presence of Mn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida |
| 13.9 | - |
mutant E122Q, presence of Mn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida |
| 140 | - |
mutant Y121A, presence of Mn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida |
| 178 | - |
wild-type, presence of EDTA, semi-apo-enzyme, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida |
| 370 | - |
wild-type, presence of Zn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida |
| 740 | - |
mutant W174F, presence of Mn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida |
| 1560 | - |
wild-type, presence of Mn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| creatinine + H2O | - |
Pseudomonas putida | creatine | - |
? | |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 1.5 | - |
creatinine | mutant E122Q, presence of Zn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida | |
| 11 | - |
creatinine | mutant E122Q, presence of Mn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida | |
| 18 | - |
creatinine | mutant W154F, presence of Mn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida | |
| 86 | - |
creatinine | mutant Y121A, presence of Mn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida | |
| 222 | - |
creatinine | wild-type, presence of EDTA, semi-apo-enzyme, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida | |
| 252 | - |
creatinine | wild-type, presence of Zn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida | |
| 688 | - |
creatinine | mutant W174F, presence of Mn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida | |
| 1340 | - |
creatinine | wild-type, presence of Mn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida | |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.008 | - |
creatinine | mutant E122Q, presence of Zn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida | |
| 0.051 | - |
creatinine | mutant W154F, presence of Mn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida | |
| 0.2 | - |
creatinine | mutant E122Q, presence of Mn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida | |
| 0.54 | - |
creatinine | mutant Y121A, presence of Mn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida | |
| 1.48 | - |
creatinine | wild-type, presence of EDTA, semi-apo-enzyme, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida | |
| 5.73 | - |
creatinine | wild-type, presence of Zn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida | |
| 7.64 | - |
creatinine | mutant W174F, presence of Mn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida | |
| 17.4 | - |
creatinine | wild-type, presence of Mn2+, pH not specified in the publication, temperature not specified in the publication | Pseudomonas putida | |
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Release 2023.1 (January 2023)
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