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Literature summary for 2.3.1.1 extracted from

  • Zhao, G.; Jin, Z.; Allewell, N.M.; Tuchman, M.; Shi, D.
    Crystal structure of the N-acetyltransferase domain of human N-acetyl-L-glutamate synthase in complex with N-acetyl-L-glutamate provides insights into its catalytic and regulatory mechanisms (2013), PLoS ONE, 8, e70369.
    View publication on PubMedView publication on EuropePMC

Activating Compound

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(Commentary)

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 (Commentary)

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

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 [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information Michaelis-Menten kinetics Homo sapiens

Localization

Localization Comment Organism GeneOntology No. Textmining
mitochondrion
-
Homo sapiens 5739
-

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
202400
-
calculated from amino acid sequence Homo sapiens
220100
-
gel filtration Homo sapiens

Natural Substrates/ Products (Substrates)

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

Organism UniProt Comment Textmining
Homo sapiens
-
-
-
Homo sapiens Q8N159
-
-

Purification (Commentary)

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 [micromol/min/mg]

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 and Products (Substrate)

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

Subunits Comment Organism
dimer catalytic N-acetyltransferase domain Homo sapiens
homotetramer
-
Homo sapiens

Synonyms

Synonyms Comment Organism
N-acetyl-L-glutamate synthase
-
Homo sapiens
NAGS
-
Homo sapiens

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
assay at Homo sapiens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.5
-
assay at Homo sapiens

General Information

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