Crystallization (Comment) | Organism |
---|---|
a 2.1 A X-ray diffraction structure of Arg166Ser alkaline phosphatase is presented, which shows little difference in enzyme structure compared to the wild-type enzyme but shows a significant reorientation of the bound phosphate | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
additional information | the results show that the role of the arginine side chain extends beyond its positive charge, as the Arg166Lys mutant is as compromised in activity as Arg166Ser. Through measurement of individual reaction steps, a free energy profile for the hydrolysis of the enzyme-phosphate intermediate is constructed. This analysis indicates that the arginine side chain strengthens binding by approximately 3 kcal/mol and provides an additional 1-2 kcal/mol stabilization of the chemical transition state | Escherichia coli |
R166A | kcat (1/sec) (substrate: 4-nitrophenyl phosphate): 0.3 | Escherichia coli |
R166K | kcat (1/sec) (substrate: 4-nitrophenyl phosphate): 0.65 | Escherichia coli |
R166S | kcat (1/sec) (substrate: 4-nitrophenyl phosphate): 0.5 | Escherichia coli |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
inorganic phosphate | - |
Escherichia coli |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | - |
Escherichia coli | |
Zn2+ | - |
Escherichia coli |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P00634 | - |
- |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
a phosphate monoester + H2O = an alcohol + phosphate | Arg166 contributes to catalysis through binding interactions and through additional transition state stabilization that may arise from complementarity of the guanidinum group to the geometry of the trigonal bipyramidal transition state | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-nitrophenyl phosphate + H2O | - |
Escherichia coli | 4-nitrophenol + phosphate | - |
? | |
ethyl phosphate + H2O | - |
Escherichia coli | ethanol + phosphate | - |
? |
Synonyms | Comment | Organism |
---|---|---|
alkaline phosphatase | - |
Escherichia coli |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kcat/KM (1/Msec) wild-type: 140000, mutant R166K: 20, mutant R166S: 24 | Escherichia coli | |
0.3 | - |
4-nitrophenyl phosphate | pH 8.0, 25°C, mutant R166A | Escherichia coli | |
0.5 | - |
4-nitrophenyl phosphate | pH 8.0, 25°C, mutant R166S | Escherichia coli | |
0.65 | - |
4-nitrophenyl phosphate | pH 8.0, 25°C, mutant R166K | Escherichia coli | |
12 | - |
4-nitrophenyl phosphate | pH 8.0, 25°C, wild-type | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Escherichia coli |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0011 | - |
inorganic phosphate | pH 8.0, 25°C, wild-type | Escherichia coli | |
0.075 | - |
inorganic phosphate | pH 8.0, 25°C, mutant R166K | Escherichia coli | |
0.46 | - |
inorganic phosphate | pH 8.0, 25°C, mutant R166S | Escherichia coli | |
0.65 | - |
inorganic phosphate | pH 8.0, 25°C, mutant R166A | Escherichia coli |