Activating Compound | Comment | Organism | Structure |
---|---|---|---|
L-arginine | allosteric activator, the binding site of the activator is located in the amino acid kinase domain of the enzyme | Homo sapiens |
Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) cells | Homo sapiens |
recombinant expression of His-tagged wild-type and mutant enzymes, and isolated catalytic N-acetyltransferase domain in Escherichia coli strain BL21(DE3) | Homo sapiens |
Crystallization (Comment) | Organism |
---|---|
catalytic N-acetyltransferase domain complexed with N-acetyl-L-glutamate, hanging drop vapor diffusion method, using 100 mM Bis-Tris, pH 6.5, 35% (w/v) PEG3350 | Homo sapiens |
purified recombinant catalytic N-acetyltransferase domain complexed with N-acetyl-L-glutamate, sitting drop vapour diffusion method, mixing of 0.002 ml of 20 mg/ml protein in 50 mM Tris-HCl, pH 7.4, 50 mM NaCl, 10% glycerol, 5 mM bmercaptoethanol, and 1 mM EDTA, 10 m CoA, and 10 mM N-acetyl-L-glutamate, with 0.002 ml of reservoir solution containing 100 mM Bis-Tris, pH 6.5, 35% PEG 3350, 18°C, X-ray diffraction structure determination and analysis at 2.1 A resolution, molecular replacement | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
additional information | the isolated catalytic N-acetyltransferase domain retains catalytic activity in the absence of the amino acid kinase domain | Homo sapiens |
N479A | the mutant shows reduced activity compared to the wild type enzyme | Homo sapiens |
N479A | site-directed mutagenesis, an active site mutant with reduced activity compared to the wild-type enzyme | Homo sapiens |
Y441F | the mutant shows reduced activity compared to the wild type enzyme | Homo sapiens |
Y441F | site-directed mutagenesis, an active site mutant with reduced activity compared to the wild-type enzyme | Homo sapiens |
Y485F | the mutant shows reduced activity compared to the wild type enzyme | Homo sapiens |
Y485F | site-directed mutagenesis, an active site mutant with reduced activity compared to the wild-type enzyme | Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten kinetics | Homo sapiens |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
mitochondrion | - |
Homo sapiens | 5739 | - |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
202400 | - |
calculated from amino acid sequence | Homo sapiens |
220100 | - |
gel filtration | Homo sapiens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
acetyl-CoA + L-glutamate | Homo sapiens | - |
CoA + N-acetyl-L-glutamate | - |
? | |
additional information | Homo sapiens | the N-acetyl-L-glutamate synthase contains an N-terminal catalytic N-acetyltransferase domain and a C-terminal amino acid kinase domain | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | - |
- |
- |
Homo sapiens | Q8N159 | - |
- |
Purification (Comment) | Organism |
---|---|
nickel affinity and Histrap column chromatography | Homo sapiens |
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and cation exchange chromatography, the His-tag is ceaved by thrombin | Homo sapiens |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.078 | - |
mutant enzyme Y485F, at pH 8.5 and 30°C | Homo sapiens |
0.154 | - |
mutant enzyme N479A, at pH 8.5 and 30°C | Homo sapiens |
0.857 | - |
mutant enzyme Y441F, at pH 8.5 and 30°C | Homo sapiens |
1.05 | - |
wild type enzyme, at pH 8.5 and 30°C | Homo sapiens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
acetyl-CoA + L-glutamate | - |
Homo sapiens | CoA + N-acetyl-L-glutamate | - |
? | |
acetyl-CoA + L-glutamate | - |
Homo sapiens | CoA + N-acetyl-L-glutamate | CoA and N-acetyl-L-glutamate binding sites structure analysis from crystal structure, detailed overview | ? | |
additional information | the N-acetyl-L-glutamate synthase contains an N-terminal catalytic N-acetyltransferase domain and a C-terminal amino acid kinase domain | Homo sapiens | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | catalytic N-acetyltransferase domain | Homo sapiens |
homotetramer | - |
Homo sapiens |
Synonyms | Comment | Organism |
---|---|---|
N-acetyl-L-glutamate synthase | - |
Homo sapiens |
NAGS | - |
Homo sapiens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.5 | - |
assay at | Homo sapiens |
General Information | Comment | Organism |
---|---|---|
malfunction | enzyme deficiency results in elevated levels of plasma ammonia which is neurotoxic | Homo sapiens |
physiological function | N-acetylglutamate synthase catalyzes the synthesis of N-acetyl-L-glutamate, an obligate cofactor for carbamyl phosphate synthetase I in the urea cycle. An N-terminal proline-rich motif of the enzyme is likely to function in signal transduction to carbamyl phosphate synthetase I, CPS1 | Homo sapiens |