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Literature summary for 2.7.2.8 extracted from

  • de Cima, S.; Gil-Ortiz, F.; Crabeel, M.; Fita, I.; Rubio, V.
    Insight on an arginine synthesis metabolon from the tetrameric structure of yeast acetylglutamate kinase (2012), PLoS ONE, 7, e34734.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli as a His-tagged fusion protein Saccharomyces cerevisiae

Crystallization (Commentary)

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

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

Inhibitors Comment Organism Structure
L-arginine
-
Saccharomyces cerevisiae

KM Value [mM]

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

Organism UniProt Comment Textmining
Saccharomyces cerevisiae Q01217
-
-

Purification (Commentary)

Purification (Comment) Organism
using Ni-NTA chromatography Saccharomyces cerevisiae

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + N-acetyl-L-glutamate
-
Saccharomyces cerevisiae ADP + N-acetyl-L-glutamate 5-phosphate
-
?

Subunits

Subunits Comment Organism
tetramer
-
Saccharomyces cerevisiae

Synonyms

Synonyms Comment Organism
yNAGK
-
Saccharomyces cerevisiae

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Saccharomyces cerevisiae

Turnover Number [1/s]

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 [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 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