Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes in Escherichia coli strain EK1104 containing the plasmid pEK695 | Escherichia coli |
Crystallization (Comment) | Organism |
---|---|
purified recombinant mutant enzyme K164E/E239K, mixing of 10 mg/ml enzyme in 50 mM Tris-acetate, pH 8.3, with 0.002 ml of crystallization buffer containing 16% w/v PEG 4000, 0.04 M Na2MoO4-2H2O, 0.04 M N-cyclohexyl-3-aminopropanesulfonic acid, and 30 mM Tris-acetate, pH 8.75, and equilibration over a reservoir of crystallization buffer of 1.0 ml, 20°C, 2 weeks, X-ray diffraction structure determination and analysis | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
D236A | site-directed mutagenesis, the wild-type enzyme shows o change in structure by SAXS through the temperature range of 4°C to 55°C, whereas the D236A ATCase exhibits a large shift toward the T state between 4°C and 30°C, with a minor shift back toward the R state between 30°C and 45°C | Escherichia coli |
K164E/E239K | site-directed mutagenesis, a mutant aspartate transcarbamoylase exists in an intermediate quaternary structure between the canonical T and R structures, crystal structure and quaternary conformation analysis, detailed overview. pH-Dependent structural alteration consistent with either a pH-induced conformational change or a pH-induced alteration in the T to R equilibrium | Escherichia coli |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-aspartate + carbamoyl phosphate | Escherichia coli | - |
phosphate + N-carbamoyl-L-aspartate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P0A786 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Escherichia coli strain EK1104 by isoelectric precipitation, anion exchange, and hydrophobic interaction chromatography, and gel filtration | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-aspartate + carbamoyl phosphate | - |
Escherichia coli | phosphate + N-carbamoyl-L-aspartate | - |
? |
Synonyms | Comment | Organism |
---|---|---|
aspartate transcarbamoylase | - |
Escherichia coli |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Escherichia coli |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
additional information | - |
the wild-type enzyme shows o change in structure by SAXS through the temperature range of 4°C to 55°C, whereas the D236A ATCase exhibits a large shift toward the T state between 4°C and 30°C, with a minor shift back toward the R state between 30°C and 45°C | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Escherichia coli |
General Information | Comment | Organism |
---|---|---|
additional information | allosteric transition between the T and R enzyme states | Escherichia coli |