Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli, pJG001 plasmid for site-directed mutagenesis, mutant Y107W expressed in dapA-negative strain AT997r- | Escherichia coli |
Crystallization (Comment) | Organism |
---|---|
mutant Y107W, hanging-drop vapor diffusion method, diffraction to beyond 2.0 A resolution, data collection and refinement statistics, solid-state structure of the mutant enzyme largely unchanged | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
Y107F | mutant, site-directed mutagenesis | Escherichia coli |
Y107W | mutant, site-directed mutagenesis | Escherichia coli |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
(S)-lysine | partial mixed inhibition with respect to (S)-aspartate 4-semialdehyde, partial non-ncompetitive inhibition with respect to pyruvate in wild-type and in Y107W mutant, Y107W mutant still retains over 25% of uninhibited activity, even at high inhibitor concentrations compared to the wild-type enzyme retaining less than 10% of normal activity | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.11 | - |
(S)-aspartate 4-semialdehyde | wild-type | Escherichia coli | |
0.16 | - |
pyruvate | Y107F mutant | Escherichia coli | |
0.26 | - |
pyruvate | wild-type | Escherichia coli | |
0.39 | - |
(S)-aspartate 4-semialdehyde | Y107W mutant | Escherichia coli | |
0.58 | - |
(S)-aspartate 4-semialdehyde | Y107F mutant | Escherichia coli | |
1.5 | - |
pyruvate | Y107W mutant | Escherichia coli |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
31290 | - |
monomer size of mutant Y107W, deduced from sequence | Escherichia coli |
31310 | - |
monomer size of wild-type, mass spectrometry | Escherichia coli |
31320 | - |
monomer size of mutant Y107W, mass spectrometry | Escherichia coli |
64000 | - |
dimer size of mutant Y107W, analytical ultracentrifugation | Escherichia coli |
123000 | - |
tetramer size of mutant Y107W, analytical ultracentrifugation | Escherichia coli |
125300 | - |
tetramer size of wild-type protein, mass spectrometry, mutant form Y107W reveals a mixture of primarily monomer and tetramer in solution | Escherichia coli |
125400 | - |
tetramer size of mutant Y107W, mass spectrometry | Escherichia coli |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P0A6L2 | - |
- |
Purification (Comment) | Organism |
---|---|
gel filtration | Escherichia coli |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
role of Tyr107 residue in determining the quaternary structure, structural, biophysical, and kinetic studies of the Y107W mutant, catalytic ability and apparent melting temperature reduced by the mutation, tetrameric quaternary structure critical to control specificity, heat stability, and intrinsic activity | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(S)-aspartate 4-semialdehyde + pyruvate | role of Tyr107 residue, located at the tight-dimer interface between two monomers, participates in a catalytic triad of residues during catalysis | Escherichia coli | dihydrodipicolinate + H2O | - |
? |
Subunits | Comment | Organism |
---|---|---|
homotetramer | wild-type, mixture of primarily monomer and tetramer in solution determined for mutant form Y107W, hydrodynamic properties of Y107W oligomers calculated by sedimentation velocity analyses | Escherichia coli |
Synonyms | Comment | Organism |
---|---|---|
DHDPS | - |
Escherichia coli |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
6.5 | - |
(S)-aspartate 4-semialdehyde | Y107W mutant | Escherichia coli | |
10.8 | - |
(S)-aspartate 4-semialdehyde | Y107F mutant | Escherichia coli | |
124 | - |
(S)-aspartate 4-semialdehyde | wild-type | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Escherichia coli |