Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli as a His-tagged fusion protein | Saccharomyces cerevisiae |
Crystallization (Comment) | Organism |
---|---|
crystal structures of both the DUF619 domain-lacking yNAGK, ligand-free as well as complexed with acetylglutamate or acetylglutamate and arginine, and of complete mature yNAGK are determined. yNAGK has as central structure a flat tetramer formed by two dimers of amino acid kinase domains | Saccharomyces cerevisiae |
Protein Variants | Comment | Organism |
---|---|---|
DELTA357-513 | truncated mutant lacking C-terminal 150 amino acids (spanning residues 38-356), belonging to the DUF619 domain family, shows that is it stabilizes yNAGK, slows catalysis and modulates feed-back inhibition by arginine. Truncated yNAGK shows doubled kcat compared to wild-type, Km is almost not affected. IC50 (L-arginine) is lowered compared to wild-type. Truncated mutand shows lower thermal stability compared to wild-type | Saccharomyces cerevisiae |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
L-arginine | - |
Saccharomyces cerevisiae |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
10 | - |
N-acetyl-L-glutamate | wild-type, pH not specified, 37°C | Saccharomyces cerevisiae | |
13.3 | - |
N-acetyl-L-glutamate | DELTA357-513 (truncated mutant lacking C-terminal 150 amino acids), pH not specified, 37°C | Saccharomyces cerevisiae | |
15.3 | - |
ATP | wild-type, pH not specified, 37°C | Saccharomyces cerevisiae | |
20.1 | - |
ATP | DELTA357-513 (truncated mutant lacking C-terminal 150 amino acids), pH not specified, 37°C | Saccharomyces cerevisiae |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Saccharomyces cerevisiae | Q01217 | - |
- |
Purification (Comment) | Organism |
---|---|
using Ni-NTA chromatography | Saccharomyces cerevisiae |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + N-acetyl-L-glutamate | - |
Saccharomyces cerevisiae | ADP + N-acetyl-L-glutamate 5-phosphate | - |
? |
Subunits | Comment | Organism |
---|---|---|
tetramer | - |
Saccharomyces cerevisiae |
Synonyms | Comment | Organism |
---|---|---|
yNAGK | - |
Saccharomyces cerevisiae |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Saccharomyces cerevisiae |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
4.9 | - |
N-acetyl-L-glutamate | wild-type, pH not specified, 37°C | Saccharomyces cerevisiae | |
7.7 | - |
ATP | wild-type, pH not specified, 37°C | Saccharomyces cerevisiae | |
8.4 | - |
N-acetyl-L-glutamate | DELTA357-513 (truncated mutant lacking C-terminal 150 amino acids), pH not specified, 37°C | Saccharomyces cerevisiae | |
13 | - |
ATP | DELTA357-513 (truncated mutant lacking C-terminal 150 amino acids), pH not specified, 37°C | Saccharomyces cerevisiae |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
400 | - |
L-arginine | DELTA357-513 (truncated mutant lacking C-terminal 150 amino acids), pH not specified, 37°C | Saccharomyces cerevisiae |
IC50 Value | IC50 Value Maximum | Comment | Organism | Inhibitor | Structure |
---|---|---|---|---|---|
0.44 | - |
DELTA357-513 (truncated mutant lacking C-terminal 150 amino acids), pH not specified, 37°C | Saccharomyces cerevisiae | L-arginine | |
0.97 | - |
wild-type, pH not specified, 37°C | Saccharomyces cerevisiae | L-arginine |