Cloned (Comment) | Organism |
---|---|
sequence comparisons, recombinant expression of wild-type and mutant enzymes | Corynebacterium pekinense |
Crystallization (Comment) | Organism |
---|---|
crystal structures analysis, overview | Corynebacterium pekinense |
Protein Variants | Comment | Organism |
---|---|---|
R169A | site-directed mutagenesis, the Km for aspartate is decreased compared to the wild-type enzyme | Corynebacterium pekinense |
R169D | site-directed mutagenesis, the mutant shows 2.57fold higher catalytic activity with aspartate than the wild-type enzyme | Corynebacterium pekinense |
R169H | site-directed mutagenesis, the mutant shows 2.13fold higher catalytic activity with aspartate than the wild-type enzyme | Corynebacterium pekinense |
R169P | site-directed mutagenesis, the mutant shows 2.25fold higher catalytic activity with aspartate than the wild-type enzyme | Corynebacterium pekinense |
R169Y | site-directed mutagenesis, the mutant shows 4.7fold higher catalytic activity with aspartate than the wild-type enzyme. The three-dimensional structure of R169Y is more stable than that of the wild-type | Corynebacterium pekinense |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
L-lysine | each dimer contains two lysine binding sites, in which one site is exclusively found in the dimer with A and B chains located at the interface between alpha and beta subunits | Corynebacterium pekinense | |
L-methionine | - |
Corynebacterium pekinense | |
L-threonine | - |
Corynebacterium pekinense | |
additional information | aspartate kinase is an allosteric enzyme, and its activity is inhibited by allosteric inhibitors, such as Lys, Met, and Thr | Corynebacterium pekinense |
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, overview | Corynebacterium pekinense |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Corynebacterium pekinense |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
18000 | - |
- |
Corynebacterium pekinense |
47000 | - |
- |
Corynebacterium pekinense |
130000 | - |
native PAGE | Corynebacterium pekinense |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-aspartate | Corynebacterium pekinense | - |
ADP + 4-phospho-L-aspartate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Corynebacterium pekinense | E0AE72 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-aspartate | - |
Corynebacterium pekinense | ADP + 4-phospho-L-aspartate | - |
? |
Subunits | Comment | Organism |
---|---|---|
heterotetramer | dimer of dimers, alpha2beta2-type structure, 2 * 47000, alpha-subunit + 2 * 18000, beta-subunit, SDS-PAGE, each dimer contains two lysine binding sites, in which one site is exclusively found in the dimer with A and B chains located at the interface between alpha and beta subunits | Corynebacterium pekinense |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
26 | - |
wild-type enzyme | Corynebacterium pekinense |
35 | - |
mutant R169Y | Corynebacterium pekinense |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
26 | - |
purified recombinant proteins, pH 8.0, half-life of the wild-type enzyme is 4.5 h, and the half-life of the mutant R169Y increases to 5.5 h | Corynebacterium pekinense |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
wild-type enzyme | Corynebacterium pekinense |
9 | - |
mutant R169Y | Corynebacterium pekinense |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
8 | - |
purified recombinant proteins, 26°C, half-life of the wild-type enzyme is 4.5 h, and the half-life of the mutant R169Y increases to 5.5 h | Corynebacterium pekinense |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Corynebacterium pekinense |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme from Corynebacterium pekinense belongs to the class II of aspartate kinases | Corynebacterium pekinense |
metabolism | aspartate kinase is the key enzyme in the biosynthesis of aspartate derived amino acids | Corynebacterium pekinense |
additional information | catalytic residue Arg169 is an important residue in substrate binding, the catalytic domain, and in inhibitor binding. Arg169 forms a hydrogen bond with Glu92, which functions as the entrance gate. R169, S172, and G171 are key substrate binding residues. Three-dimensional structure analysis and structure homology modelling, overview | Corynebacterium pekinense |
physiological function | aspartate kinase is a multimer and an allosteric enzyme that catalyzes the phosphorylation of aspartate to form aspartyl phosphate. Allosteric regulation (feedback inhibition) of proteins controls the synthesis pathway of this enzyme | Corynebacterium pekinense |