Information on EC 2.7.2.8 - acetylglutamate kinase

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The expected taxonomic range for this enzyme is: Eukaryota, Bacteria, Archaea

EC NUMBER
COMMENTARY
2.7.2.8
-
RECOMMENDED NAME
GeneOntology No.
acetylglutamate kinase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
ATP + N-acetyl-L-glutamate = ADP + N-acetyl-L-glutamyl 5-phosphate
show the reaction diagram
-
-
-
-
ATP + N-acetyl-L-glutamate = ADP + N-acetyl-L-glutamyl 5-phosphate
show the reaction diagram
enzyme has to interact stoichiometrically with acetylglutamate synthase in order to be active
-
ATP + N-acetyl-L-glutamate = ADP + N-acetyl-L-glutamyl 5-phosphate
show the reaction diagram
random bi-bi mechanism
-
ATP + N-acetyl-L-glutamate = ADP + N-acetyl-L-glutamyl 5-phosphate
show the reaction diagram
mechanism
-
ATP + N-acetyl-L-glutamate = ADP + N-acetyl-L-glutamyl 5-phosphate
show the reaction diagram
using an elastic network model representation a normal mode analysis shows that the conformational mechanisms for substrate binding by NAGK strongly correlate with the intrinsic dynamics of the enzyme in the unbound form. The conformational change observed between the open and closed forms of EcNAGK are essentially accomplished by movements along a small subset of modes
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Acyl group transfer
Q8P8J7
-
phospho group transfer
-
-
-
-
phospho group transfer
Q8P8J7
-
phospho group transfer
-
-
phospho group transfer
-
-
phospho group transfer
Q9X2A4, -
-
PATHWAY
KEGG Link
MetaCyc Link
Arginine and proline metabolism
-
arginine biosynthesis II (acetyl cycle)
-
arginine biosynthesis III (via N-acetyl-L-citrulline)
-
arginine biosynthesis IV (archaebacteria)
-
Biosynthesis of secondary metabolites
-
Metabolic pathways
-
ornithine biosynthesis
-
SYSTEMATIC NAME
IUBMB Comments
ATP:N-acetyl-L-glutamate 5-phosphotransferase
-
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
acetylglutamate kinase
Q8LA25
-
acetylglutamate kinase
Q9SCL7
-
acetylglutamate kinase
Q9HTN2
-
acetylglutamate kinase
Q9X2A4
-
acetylglutamate kinase
Q8P8J7
-
acetylglutamate phosphokinase
-
-
-
-
amino-acid acetyltransferase
Q8P8J7
-
argB
-
gene name
EcNAGK
C6EE50
-
kinase, acetylglutamate (phosphorylating)
-
-
-
-
N-acetyl-glutamate 5-phosphotransferase
-
-
N-acetyl-L-glutamate 5-phosphotransferase
Q9HTN2
-
N-acetyl-L-glutamate 5-phosphotransferase
Q9X2A4
-
N-acetyl-L-glutamate kinase
-
-
N-acetyl-L-glutamate kinase
-
-
N-acetyl-L-glutamate kinase
-
-
N-acetylglutamate 5-phosphotransferase
-
-
-
-
N-acetylglutamate kinase
-
-
-
-
N-acetylglutamate kinase
Q8LA25
-
N-acetylglutamate kinase
Q9SCL7
-
N-acetylglutamate kinase
-
-
N-acetylglutamate kinase
Q9HTN2
-
N-acetylglutamate kinase
-
-
N-acetylglutamate kinase
Q9X2A4
-
N-acetylglutamate kinase
Q8P8J7
-
N-acetylglutamate phosphokinase
-
-
-
-
N-acetylglutamate-5-phosphotransferase
-
-
-
-
N-acetylglutamic 5-phosphotransferase
-
-
-
-
NagK
Q9X2A4
-
NAGK1
-
-
NAGS-K
Q8LA25
-
NAGS-K
Q9SCL7
-
NAGS-K
-
enzyme has both N-acetylglutamate synthase and kinases activity
NAGS-K
Q9HTN2
-
NAGS-K
Q9X2A4
-
CAS REGISTRY NUMBER
COMMENTARY
9027-58-1
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
-
Q8LA25
SwissProt
Manually annotated by BRENDA team
ATCC 25542
-
-
Manually annotated by BRENDA team
strain W2D, ATCC 25542, derepressed mutant
-
-
Manually annotated by BRENDA team
strain Wc2
-
-
Manually annotated by BRENDA team
Escherichia coli W2D
strain W2D, ATCC 25542, derepressed mutant
-
-
Manually annotated by BRENDA team
strain Wc2
-
-
Manually annotated by BRENDA team
recombinant enzyme
-
-
Manually annotated by BRENDA team
Sulfolobus solfataricus P1
-
-
-
Manually annotated by BRENDA team
strain PCC 7942, recombinant protein with His6-tag
-
-
Manually annotated by BRENDA team
recombinant enzyme
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
malfunction
-
argB gene coding the N-acetyl-L-glutamate kinase is first overexpressed in the strain SYPA5-5, whereas the L-arginine production is narrowly increased by 15.4%
metabolism
-
Sulfolobus solfataricus lacks ornithine acetyltransferase and thus forms N-acetylglutamate exclusively via the energetically less favourable reaction catalysed by N-acetylglutamate synthase, investing 1 mol of acetyl CoA per mol of N-acetyl intermediate synthesized
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
acetyl-CoA + L-glutamate
coenzyme A + N-acetyl-L-glutamate
show the reaction diagram
Q8P8J7
-
ancestral bifunctional N-acetylglutamate synthase and kinase
-
?
acetyl-CoA + L-glutamate
coenzyme A + N-acetyl-L-glutamate
show the reaction diagram
-
-
ancestral bifunctional N-acetylglutamate synthase and kinase
-
?
ATP + N-acetyl-L-glutamate
ADP + N-acetyl-L-glutamate 5-phosphate
show the reaction diagram
-
-
-
-
ATP + N-acetyl-L-glutamate
ADP + N-acetyl-L-glutamate 5-phosphate
show the reaction diagram
-
-
-
-
-
ATP + N-acetyl-L-glutamate
ADP + N-acetyl-L-glutamate 5-phosphate
show the reaction diagram
-
-
-
-
?
ATP + N-acetyl-L-glutamate
ADP + N-acetyl-L-glutamate 5-phosphate
show the reaction diagram
-
-
-
?
ATP + N-acetyl-L-glutamate
ADP + N-acetyl-L-glutamate 5-phosphate
show the reaction diagram
-
-
-
-
?
ATP + N-acetyl-L-glutamate
ADP + N-acetyl-L-glutamate 5-phosphate
show the reaction diagram
-
-
-
-
?
ATP + N-acetyl-L-glutamate
ADP + N-acetyl-L-glutamate 5-phosphate
show the reaction diagram
-
-
-
-
-
ATP + N-acetyl-L-glutamate
ADP + N-acetyl-L-glutamate 5-phosphate
show the reaction diagram
-
-
-
-
?
ATP + N-acetyl-L-glutamate
ADP + N-acetyl-L-glutamate 5-phosphate
show the reaction diagram
-
-
-
-
?
ATP + N-acetyl-L-glutamate
ADP + N-acetyl-L-glutamate 5-phosphate
show the reaction diagram
-
-
-
-
-
ATP + N-acetyl-L-glutamate
ADP + N-acetyl-L-glutamate 5-phosphate
show the reaction diagram
-
-
-
-
-
ATP + N-acetyl-L-glutamate
ADP + N-acetyl-L-glutamate 5-phosphate
show the reaction diagram
-
-
-
-
?
ATP + N-acetyl-L-glutamate
ADP + N-acetyl-L-glutamate 5-phosphate
show the reaction diagram
Q8LA25
-
-
-
-
ATP + N-acetyl-L-glutamate
ADP + N-acetyl-L-glutamate 5-phosphate
show the reaction diagram
-
-
-
-
-
ATP + N-acetyl-L-glutamate
ADP + N-acetyl-L-glutamate 5-phosphate
show the reaction diagram
Q8P8J7
-
-
-
?
ATP + N-acetyl-L-glutamate
ADP + N-acetyl-L-glutamate 5-phosphate
show the reaction diagram
-
-
-
-
?
ATP + N-acetyl-L-glutamate
ADP + N-acetyl-L-glutamate 5-phosphate
show the reaction diagram
Q9SCL7
-
-
-
?
ATP + N-acetyl-L-glutamate
ADP + N-acetyl-L-glutamate 5-phosphate
show the reaction diagram
Q9X2A4, -
-
-
-
?
ATP + N-acetyl-L-glutamate
ADP + N-acetyl-L-glutamate 5-phosphate
show the reaction diagram
-, Q9HTN2
-
-
-
?
ATP + N-acetyl-L-glutamate
ADP + N-acetyl-L-glutamate 5-phosphate
show the reaction diagram
-
-
-
-
?
ATP + N-acetyl-L-glutamate
ADP + N-acetyl-L-glutamate 5-phosphate
show the reaction diagram
-
-
-
-
?
ATP + N-acetyl-L-glutamate
ADP + N-acetyl-L-glutamate 5-phosphate
show the reaction diagram
Q01217, -
-
-
-
?
ATP + N-acetyl-L-glutamate
ADP + N-acetyl-L-glutamate 5-phosphate
show the reaction diagram
C6EE50, -
-
-
-
?
ATP + N-acetyl-L-glutamate
ADP + N-acetyl-L-glutamate 5-phosphate
show the reaction diagram
-
-
in presence of hydroxylamine formation of N-acetyl-L-glutamate 5-hydroxamate + ADP + phosphate
?
ATP + N-acetyl-L-glutamate
ADP + N-acetyl-L-glutamate 5-phosphate
show the reaction diagram
-
-
in presence of hydroxylamine formation of N-acetyl-L-glutamate 5-hydroxamate + ADP + phosphate
?
ATP + N-acetyl-L-glutamate
ADP + N-acetyl-L-glutamate 5-phosphate
show the reaction diagram
-
highly specific for ATP and N-acetyl-L-glutamate
in presence of hydroxylamine formation of N-acetyl-L-glutamate 5-hydroxamate + ADP + phosphate
?
ATP + N-acetyl-L-glutamate
ADP + N-acetyl-L-glutamate 5-phosphate
show the reaction diagram
-
key enzyme in regulation of arginine biosynthesis
-
-
ATP + N-acetyl-L-glutamate
ADP + N-acetyl-L-glutamate 5-phosphate
show the reaction diagram
-
key enzyme in regulation of arginine biosynthesis
-
-
ATP + N-acetyl-L-glutamate
ADP + N-acetyl-L-glutamate 5-phosphate
show the reaction diagram
-
enzyme synthesis not repressed by exogenous L-arginine or its precursors, second enzyme of arginine biosynthesis
-
-
-
ATP + N-acetyl-L-glutamate
ADP + N-acetyl-L-glutamate 5-phosphate
show the reaction diagram
Escherichia coli W2D
-
-
-
-, ?
ATP + N-acetyl-L-glutamate
ADP + N-acetyl-L-glutamate 5-phosphate
show the reaction diagram
-
-
-
-
?
ATP + N-acetyl-L-glutamate
ADP + N-acetyl-L-glutamyl 5-phosphate
show the reaction diagram
-
-
-
-
?
ATP + N-acetyl-L-glutamate
ADP + N-acetyl-L-glutamyl 5-phosphate
show the reaction diagram
-
-
-
-
?
ATP + N-acetyl-L-glutamate
ADP + N-acetyl-L-glutamyl 5-phosphate
show the reaction diagram
-
NAGK catalyzes the second step of arginine biosynthesis. In Pseudomonas aeruginosa, this step is rate limiting, and feedback regulated and sigmoidally inhibited by arginine
-
-
?
ATP + N-carbamoyl-L-glutamate
ADP + N-carbamoyl-L-glutamate 5-phosphate
show the reaction diagram
-
at 33% of the activity with N-acetyl-L-glutamate
-
-
-
ATP + N-formyl-L-glutamate
ADP + N-formyl-L-glutamate 5-phosphate
show the reaction diagram
-
at 20% of the activity with N-acetyl-L-glutamate
-
-
-
dATP + N-acetyl-L-glutamate
dADP + N-acetyl-L-glutamate 5-phosphate
show the reaction diagram
-
as effective as ATP
-
-
-
additional information
?
-
-
not: ITP
-
-
-
additional information
?
-
-
not: GTP
-
-
-
additional information
?
-
-
not: GTP
-
-
-
additional information
?
-
-
not: GTP
-
-
-
additional information
?
-
-
not: N-benzoyl-L-glutamate, L-glutamate, D-glutamate
-
-
-
additional information
?
-
-
not: N-benzoyl-L-glutamate, L-glutamate, D-glutamate
-
-
-
additional information
?
-
-
N-propionyl-L-glutamate almost inactive
-
-
-
additional information
?
-
-
NAGK strongly interacts with PII protein only in the presence of Mg-ATP, this process is reversed by 2-oxoglutarate
-
-
-
additional information
?
-
-
NAGK interacts with PII protein
-
-
-
additional information
?
-
Escherichia coli W2D
-
not: GTP, not: N-benzoyl-L-glutamate, L-glutamate, D-glutamate
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
ATP + N-acetyl-L-glutamate
ADP + N-acetyl-L-glutamate 5-phosphate
show the reaction diagram
-
-
-
-
ATP + N-acetyl-L-glutamate
ADP + N-acetyl-L-glutamate 5-phosphate
show the reaction diagram
-
-
-
-
?
ATP + N-acetyl-L-glutamate
ADP + N-acetyl-L-glutamate 5-phosphate
show the reaction diagram
-
-
-
-
?
ATP + N-acetyl-L-glutamate
ADP + N-acetyl-L-glutamate 5-phosphate
show the reaction diagram
-
-
-
-
?
ATP + N-acetyl-L-glutamate
ADP + N-acetyl-L-glutamate 5-phosphate
show the reaction diagram
-
key enzyme in regulation of arginine biosynthesis
-
-
ATP + N-acetyl-L-glutamate
ADP + N-acetyl-L-glutamate 5-phosphate
show the reaction diagram
-
key enzyme in regulation of arginine biosynthesis
-
-
ATP + N-acetyl-L-glutamate
ADP + N-acetyl-L-glutamate 5-phosphate
show the reaction diagram
-
enzyme synthesis not repressed by exogenous L-arginine or its precursors, second enzyme of arginine biosynthesis
-
-
-
ATP + N-acetyl-L-glutamate
ADP + N-acetyl-L-glutamyl 5-phosphate
show the reaction diagram
-
NAGK catalyzes the second step of arginine biosynthesis. In Pseudomonas aeruginosa, this step is rate limiting, and feedback regulated and sigmoidally inhibited by arginine
-
-
?
ATP + N-acetyl-L-glutamate
ADP + N-acetyl-L-glutamate 5-phosphate
show the reaction diagram
Escherichia coli W2D
-
-
-
-
additional information
?
-
-
NAGK strongly interacts with PII protein only in the presence of Mg-ATP, this process is reversed by 2-oxoglutarate
-
-
-
additional information
?
-
-
NAGK interacts with PII protein
-
-
-
COFACTOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Ca2+
-
Mn2+, Zn2+, Co2+ and Ca2+ in this order can partially replace Mg2+
Co2+
-
Mg2+ or Co2+ required
Co2+
-
Mn2+, Zn2+, Co2+ and Ca2+ in this order can partially replace Mg2+
Mg2+
-
employed in assay mixture
Mg2+
-
Mg2+ or Co2+ required
Mg2+
-
Mg2+ or Co2+ required; required
Mg2+
-
required
Mg2+
-
30 mM, slight inhibition; required
Mg2+
-
required, maximum activity above 20 mM
Mg2+
-
required
Mn2+
-
less effective than Mg2+ and Co2+ in activation
Mn2+
-
Mn2+, Zn2+, Co2+ and Ca2+ in decreasing order can partially replace Mg2+
Zn2+
-
Mn2+, Zn2+, Co2+ and Ca2+ in this order can partially replace Mg2+
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
2-oxoglutarate
-
complete inhibition at 0.5 mM
ADP
-
inhibitor of complex formation with PII protein
arginine
-
feedback regulation of enzyme and N-acetylglutamate synthase is mutually interdependent, enzymes form a complex
arginine
-
pH-optimum for inhibition is 7.5
arginine
-
allosteric enzyme which is inhibited by arginine
arginine
-
enzyme has no allosteric properties and its activity is influenced neither by arginine nor by any of the intermediates of the arginine biosynthetic pathway
arginine
-
feedback inhibition is markedly dependent on pH, above pH 9 no inhibition
arginine
-
feedback inhibition is markedly dependent on pH, above pH 9 no inhibition
arginine
-
1 mM, 96% inhibition
arginine
-
feedback inhibition
arginine
Q9X2A4, -
-
L-arginine
-
pH and temperature dependent, sigmoidal dependence on concentration of arginine, Hill coefficient of 4, 1 mM, 37C, 95% inhibition
L-arginine
-
only for N-acetylglutamate synthase activity
L-arginine
-
less inhibitory in the presence of the PII protein
L-arginine
-
sigmoidal arginine inhibition kinetics, feedback inhibition, indentification of the N-terminal arginine site, mutational analysis, the mobile alphaH-beta16 loop of the arginine site is the modulatory signal receiver, overview
L-arginine
-
complete inhibition at 3-5 mM, PII-mediated relief from L-arginine inhibition is antagonized by 2-oxoglutarate
L-arginine
-
complete inhibition at 0.1-1.0 mM, PII-mediated relief from L-arginine inhibition is antagonized by 2-oxoglutarate
L-arginine
Q01217, -
-
L-arginine methyl ester
-
1 mM, 80% inhibition
MgCl2
-
inhibition above 20 mM
MgCl2
-
30 mM, slight inhibition
additional information
-
not inhibitory: D-arginine, agmatine, citrulline, L-canavanine, L-lysine, L-ornithine, guanidinium ions, urea at 1 mM, 37C
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
2-Ketoglutarate
-
only if bound to the PII protein ATP complex
ATP
-
maximum activity at 10 mM
PII protein
-
prevents inhibition by arginine
-
PII protein
-
-
-
PII protein
-
-
-
L-arginine
-
slight activation is observed at 1 mM L-arginine
additional information
-
2-ketoglutarate shows no additive effect to the activation of PII protein
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
1.3
-
AcCoA
-
only for N-acetylglutamate synthase activity
0.29
-
ATP
-
wild-type, pH 7.0, 25C
0.3
-
ATP
-
in the absence of protein PII, in 50 mM imidazole, pH 7.5
0.4
-
ATP
-
in the absence of protein PII, in 50 mM imidazole, pH 7.5
0.46
-
ATP
-
mutant D162E, pH 7.0, 25C
0.6
-
ATP
-
in the absence of protein PII, in 50 mM imidazole, pH 7.5
1.1
-
ATP
-
in the absence of protein PII, in 50 mM imidazole, pH 7.5
1.1
-
ATP
C6EE50, -
pH 7.5, 37C, wild-type, Vmax: 72.6
1.6
-
ATP
-
pH 5.5, 37C
1.7
-
ATP
-
pH 7.5, 30C
1.74
-
ATP
-
without PII protein
2.03
-
ATP
-
in complex with PII protein
2.47
-
ATP
-
in complex with PII protein and 2-ketoglutarate
3
-
ATP
-
pH 8.0, 37C, allosteric
3
-
ATP
-
pH 7.2, 37C
3.1
-
ATP
-
wild-type enzyme
3.3
-
ATP
-
mutant K8R, pH 7.0, 25C
5.2
-
ATP
-
mutant R66K, pH 7.0, 25C
7.9
-
ATP
-
mutant N158Q, pH 7.0, 25C
13.4
-
ATP
C6EE50, -
pH 7.5, 37C, mutant G11A, Vmax: 11.1
15.3
-
ATP
Q01217, -
wild-type, pH not specified, 37C
20.1
-
ATP
Q01217, -
DELTA357-513 (truncated mutant lacking C-terminal 150 amino acids), pH not specified, 37C
0.2
-
N-acetyl-L-glutamate
-
wild-type, pH 7.0, 25C
0.37
-
N-acetyl-L-glutamate
-
mutant D162E, pH 7.0, 25C
0.85
-
N-acetyl-L-glutamate
-
in the absence of protein PII, in 50 mM imidazole, pH 7.5
0.87
-
N-acetyl-L-glutamate
-
in the absence of protein PII, in 50 mM imidazole, pH 7.5
1.3
-
N-acetyl-L-glutamate
C6EE50, -
pH 7.5, 37C, wild-type, Vmax: 80
1.9
-
N-acetyl-L-glutamate
-
two Km-values: 1.9 and 6.2, pH 7.5, 30C
2
-
N-acetyl-L-glutamate
-
pH 7.2, 37C
2.3
-
N-acetyl-L-glutamate
-
mutant K8R, pH 7.0, 25C
2.7
-
N-acetyl-L-glutamate
-
pH 7.5, complex of enzyme and PII protein
3.1
-
N-acetyl-L-glutamate
-
wild-type enzyme
3.1
-
N-acetyl-L-glutamate
-
in the absence of protein PII, in 50 mM imidazole, pH 7.5
3.14
-
N-acetyl-L-glutamate
-
mutant H26A, pH and temperature not specified in the publication
3.25
-
N-acetyl-L-glutamate
-
mutant H268N, pH and temperature not specified in the publication
3.29
-
N-acetyl-L-glutamate
-
mutant H26E, pH 8.0, 37C
3.32
-
N-acetyl-L-glutamate
-
mutant G287D, pH and temperature not specified in the publication
3.33
-
N-acetyl-L-glutamate
-
mutant E19R, pH 8.0, 37C
3.36
-
N-acetyl-L-glutamate
-
mutant H268N/H26E/E19R, pH 8.0, 37C
3.42
-
N-acetyl-L-glutamate
-
mutant H268N/H26E, pH 8.0, 37C; wild-type, pH 8.0, 37C
3.43
-
N-acetyl-L-glutamate
-
mutant E19R, pH and temperature not specified in the publication
3.45
-
N-acetyl-L-glutamate
-
mutant H268N, pH 8.0, 37C
3.52
-
N-acetyl-L-glutamate
-
wild-type, pH and temperature not specified in the publication
3.61
-
N-acetyl-L-glutamate
-
mutant E19A, pH and temperature not specified in the publication
3.72
-
N-acetyl-L-glutamate
-
mutant R209A, pH and temperature not specified in the publication
3.76
-
N-acetyl-L-glutamate
-
mutant H268A, pH and temperature not specified in the publication
3.86
-
N-acetyl-L-glutamate
-
mutant G287A, pH and temperature not specified in the publication
3.89
-
N-acetyl-L-glutamate
-
mutant W23A, pH and temperature not specified in the publication
3.96
-
N-acetyl-L-glutamate
-
mutant R209K, pH and temperature not specified in the publication
4
-
N-acetyl-L-glutamate
-
pH 8.0, 37C, allosteric
5.1
-
N-acetyl-L-glutamate
C6EE50, -
pH 7.5, 37C, mutant G11A, Vmax: 9.7
6
-
N-acetyl-L-glutamate
-
pH 5.5, 37C
6.2
-
N-acetyl-L-glutamate
-
2 Km-values: 1.9 and 6.2, pH 7.5, 30C
6.98
-
N-acetyl-L-glutamate
-
mutant DELTA8, pH and temperature not specified in the publication
7.08
-
N-acetyl-L-glutamate
-
without PII protein
7.4
-
N-acetyl-L-glutamate
-
in the absence of protein PII, in 50 mM imidazole, pH 7.5
7.55
-
N-acetyl-L-glutamate
-
in complex with PII protein
7.84
-
N-acetyl-L-glutamate
-
mutant DELTA2-15, pH and temperature not specified in the publication
8.23
-
N-acetyl-L-glutamate
-
mutant DELTA2-19, pH and temperature not specified in the publication
8.45
-
N-acetyl-L-glutamate
-
mutant DELTA2-29, pH and temperature not specified in the publication
9.45
-
N-acetyl-L-glutamate
-
in complex with PII protein and 2-ketoglutarate
10
-
N-acetyl-L-glutamate
Q01217, -
wild-type, pH not specified, 37C
13.3
-
N-acetyl-L-glutamate
Q01217, -
DELTA357-513 (truncated mutant lacking C-terminal 150 amino acids), pH not specified, 37C
13.68
-
N-acetyl-L-glutamate
-
mutant E281A, pH and temperature not specified in the publication
15
-
N-acetyl-L-glutamate
-
pH 5.5, 37C
15
-
N-acetyl-L-glutamate
-
-
25.74
-
N-acetyl-L-glutamate
-
mutant DELTA25, pH and temperature not specified in the publication
27.4
-
N-acetyl-L-glutamate
-
pH 7.5, free enzyme
85
-
N-acetyl-L-glutamate
-
in the presence of 1 mM L-arginine, in 50 mM imidazole, pH 7.5
600
-
N-acetyl-L-glutamate
-
mutant N158Q, pH 7.0, 25C
898
-
N-acetyl-L-glutamate
-
mutant R66K, pH 7.0, 25C
2.8
-
L-glutamate
-
only for N-acetylglutamate synthase activity
additional information
-
additional information
-
formation of complex between enzyme and signal transduction protein PII decreases Km-value by a factor of 10 and increases Vmax 4fold
-
additional information
-
additional information
-
kinetics of enzyme mutants, overview
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
7.7
-
ATP
Q01217, -
wild-type, pH not specified, 37C
13
-
ATP
Q01217, -
DELTA357-513 (truncated mutant lacking C-terminal 150 amino acids), pH not specified, 37C
2
-
N-acetyl-L-glutamate
-
mutant DELTA25, pH and temperature not specified in the publication, with 1 mM L-arginine
3.2
-
N-acetyl-L-glutamate
-
mutant DELTA25, pH and temperature not specified in the publication
4.2
-
N-acetyl-L-glutamate
-
mutant DELTA8, pH and temperature not specified in the publication, with 1 mM L-arginine
4.9
-
N-acetyl-L-glutamate
Q01217, -
wild-type, pH not specified, 37C
6.4
-
N-acetyl-L-glutamate
-
mutant DELTA8, pH and temperature not specified in the publication
7.2
-
N-acetyl-L-glutamate
-
mutant DELTA2-29, pH and temperature not specified in the publication, with 1 mM L-arginine
7.9
-
N-acetyl-L-glutamate
-
mutant DELTA2-29, pH and temperature not specified in the publication
8.4
-
N-acetyl-L-glutamate
Q01217, -
DELTA357-513 (truncated mutant lacking C-terminal 150 amino acids), pH not specified, 37C
11.5
-
N-acetyl-L-glutamate
-
mutant DELTA2-19, pH and temperature not specified in the publication, with 1 mM L-arginine
12.4
-
N-acetyl-L-glutamate
-
mutant DELTA2-19, pH and temperature not specified in the publication
13.1
-
N-acetyl-L-glutamate
-
in the absence of protein PII, in 50 mM imidazole, pH 7.5
13.8
-
N-acetyl-L-glutamate
-
mutant DELTA2-15, pH and temperature not specified in the publication, with 1 mM L-arginine
15.2
-
N-acetyl-L-glutamate
-
mutant DELTA2-15, pH and temperature not specified in the publication
31.4
-
N-acetyl-L-glutamate
-
wild-type, pH 8.0, 37C, with 1 mM L-arginine
31.4
-
N-acetyl-L-glutamate
-
wild-type, pH and temperature not specified in the publication, with 1 mM L-arginine
32.8
-
N-acetyl-L-glutamate
-
mutant W23A, pH and temperature not specified in the publication, with 1 mM L-arginine
34.9
-
N-acetyl-L-glutamate
-
mutant E281A, pH and temperature not specified in the publication, with 1 mM L-arginine
36.1
-
N-acetyl-L-glutamate
-
mutant G287A, pH and temperature not specified in the publication, with 1 mM L-arginine
36.7
-
N-acetyl-L-glutamate
-
mutant G287A, pH and temperature not specified in the publication
37.4
-
N-acetyl-L-glutamate
-
mutant R209K, pH and temperature not specified in the publication, with 1 mM L-arginine
38.3
-
N-acetyl-L-glutamate
-
mutant G287D, pH and temperature not specified in the publication, with 1 mM L-arginine
38.5
-
N-acetyl-L-glutamate
-
mutant E19A, pH and temperature not specified in the publication, with 1 mM L-arginine
39.7
-
N-acetyl-L-glutamate
-
mutant R209A, pH and temperature not specified in the publication, with 1 mM L-arginine; mutant W23A, pH and temperature not specified in the publication
40.4
-
N-acetyl-L-glutamate
-
in the presence of protein PII, in 50 mM imidazole, pH 7.5
41
-
N-acetyl-L-glutamate
-
mutant H26E, pH 8.0, 37C, with 1 mM L-arginine
41
-
N-acetyl-L-glutamate
-
mutant E281A, pH and temperature not specified in the publication
41.1
-
N-acetyl-L-glutamate
-
mutant H268N, pH 8.0, 37C, with 1 mM L-arginine
41.1
-
N-acetyl-L-glutamate
-
mutant H268A, pH and temperature not specified in the publication, with 1 mM L-arginine
41.3
-
N-acetyl-L-glutamate
-
mutant G287D, pH and temperature not specified in the publication
41.9
-
N-acetyl-L-glutamate
-
mutant R209A, pH and temperature not specified in the publication
42
-
N-acetyl-L-glutamate
-
mutant E19R, pH 8.0, 37C, with 1 mM L-arginine
42.1
-
N-acetyl-L-glutamate
-
mutant E19R, pH and temperature not specified in the publication, with 1 mM L-arginine; mutant R209K, pH and temperature not specified in the publication
43
-
N-acetyl-L-glutamate
-
mutant H26A, pH and temperature not specified in the publication, with 1 mM L-arginine
43.3
-
N-acetyl-L-glutamate
-
mutant H268N, pH and temperature not specified in the publication
43.4
-
N-acetyl-L-glutamate
-
mutant H26A, pH and temperature not specified in the publication
43.8
-
N-acetyl-L-glutamate
-
mutant H268N, pH and temperature not specified in the publication, with 1 mM L-arginine
44.1
-
N-acetyl-L-glutamate
-
mutant E19A, pH and temperature not specified in the publication
44.4
-
N-acetyl-L-glutamate
-
mutant H26E, pH 8.0, 37C
44.5
-
N-acetyl-L-glutamate
-
mutant H268A, pH and temperature not specified in the publication
44.8
-
N-acetyl-L-glutamate
-
mutant E19R, pH and temperature not specified in the publication
45.1
-
N-acetyl-L-glutamate
-
wild-type, pH 8.0, 37C
45.1
-
N-acetyl-L-glutamate
-
wild-type, pH and temperature not specified in the publication
45.3
-
N-acetyl-L-glutamate
-
mutant H268N, pH 8.0, 37C
45.6
-
N-acetyl-L-glutamate
-
mutant H268N/H26E, pH 8.0, 37C, with 1 mM L-arginine
45.8
-
N-acetyl-L-glutamate
-
mutant E19R, pH 8.0, 37C
46
-
N-acetyl-L-glutamate
-
mutant H268N/H26E/E19R, pH 8.0, 37C, with 1 mM L-arginine
46.1
-
N-acetyl-L-glutamate
-
mutant H268N/H26E, pH 8.0, 37C
46.5
-
N-acetyl-L-glutamate
-
mutant H268N/H26E/E19R, pH 8.0, 37C
115
-
N-acetyl-L-glutamate
-
in the presence of 1 mM L-arginine, in 50 mM imidazole, pH 7.5
126.1
-
N-acetyl-L-glutamate
-
in the absence of protein PII, in 50 mM imidazole, pH 7.5
187.5
-
N-acetyl-L-glutamate
-
in the presence of protein PII, in 50 mM imidazole, pH 7.5
kcat/KM VALUE [1/mMs-1]
kcat/KM VALUE [1/mMs-1] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
1.3
-
N-acetyl-L-glutamate
-
in the presence of 1 mM L-arginine, in 50 mM imidazole, pH 7.5
13560
1.7
-
N-acetyl-L-glutamate
-
in the absence of protein PII, in 50 mM imidazole, pH 7.5
13560
13
-
N-acetyl-L-glutamate
-
in the presence of protein PII, in 50 mM imidazole, pH 7.5
13560
145
-
N-acetyl-L-glutamate
-
in the absence of protein PII, in 50 mM imidazole, pH 7.5
13560
220
-
N-acetyl-L-glutamate
-
in the presence of protein PII, in 50 mM imidazole, pH 7.5
13560
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
30.4
-
ATP
-
wild-type, pH 7.0, 25C
41.5
-
ATP
-
mutant N158Q, pH 7.0, 25C
400
-
L-arginine
Q01217, -
DELTA357-513 (truncated mutant lacking C-terminal 150 amino acids), pH not specified, 37C
IC50 VALUE [mM]
IC50 VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.02
-
L-arginine
-
in the absence of protein PII, in 50 mM imidazole, pH 7.5
0.19
-
L-arginine
-
in the presence of protein PII, in 50 mM imidazole, pH 7.5
0.4
-
L-arginine
-
wild-type, pH 8.0, 37C
0.4
-
L-arginine
-
wild-type, pH and temperature not specified in the publication
0.44
-
L-arginine
Q01217, -
DELTA357-513 (truncated mutant lacking C-terminal 150 amino acids), pH not specified, 37C
0.97
-
L-arginine
Q01217, -
wild-type, pH not specified, 37C
1
-
L-arginine
-
in the absence of protein PII, in 50 mM imidazole, pH 7.5
1.8
-
L-arginine
-
mutant W23A, pH and temperature not specified in the publication
5.9
-
L-arginine
-
in the presence of protein PII, in 50 mM imidazole, pH 7.5
6.4
-
L-arginine
-
mutant DELTA8, pH and temperature not specified in the publication
6.8
-
L-arginine
-
mutant DELTA25, pH and temperature not specified in the publication
8.6
-
L-arginine
-
mutant E281A, pH and temperature not specified in the publication
14.8
-
L-arginine
-
mutant R209K, pH and temperature not specified in the publication
14.9
-
L-arginine
-
mutant G287A, pH and temperature not specified in the publication
15.7
-
L-arginine
-
mutant H268A, pH and temperature not specified in the publication
16.1
-
L-arginine
-
mutant E19A, pH and temperature not specified in the publication
16.2
-
L-arginine
-
mutant G287D, pH and temperature not specified in the publication
18.5
-
L-arginine
-
mutant R209A, pH and temperature not specified in the publication
21
-
L-arginine
-
mutant H26E, pH 8.0, 37C
21.7
-
L-arginine
-
mutant H26A, pH and temperature not specified in the publication
22
-
L-arginine
-
mutant H268N, pH 8.0, 37C
22.3
-
L-arginine
-
mutant H268N, pH and temperature not specified in the publication
24
-
L-arginine
-
mutant E19R, pH 8.0, 37C
24.4
-
L-arginine
-
mutant E19R, pH and temperature not specified in the publication
28.4
-
L-arginine
-
mutant DELTA2-15, pH and temperature not specified in the publication
59.2
-
L-arginine
-
mutant DELTA2-19, pH and temperature not specified in the publication
65.2
-
L-arginine
-
mutant DELTA2-29, pH and temperature not specified in the publication
350
-
L-arginine
-
mutant H268N/H26E, pH 8.0, 37C
784
-
L-arginine
-
mutant H268N/H26E/E19R, pH 8.0, 37C
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0.0008
-
-
pH 8.0, 70C, activity in cell culture in growth medium with 0.2% yeast extract
0.003
-
-
pH 8.0, 70C, activity in cell culture in minimal growth medium with 20 mM glucose and 5 mM NH4+
0.54
-
-
pH 7.4, 37C
3.3
-
-
pH 7.5, free enzyme
12
-
-
without PII protein
13.3
-
-
pH 7.5, complex of enzyme and PII protein
20.1
-
-
pH 7.4, 37C
24
-
-
with PII protein
45
-
-
37C, pH 7.5
130
-
-
37C, pH 7.5
677
-
-
80C, pH 7.5
additional information
-
-
-
additional information
-
-
-
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7.4
-
-
assay at
7.5
-
-
both free enzyme and complex with protein PII
7.5
-
-
assay at
7.5
-
C6EE50, -
assay at
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4.6
7
-
pH 4.6: about 75% of maximum activity, pH 7: about 60% of maximum activity
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
25
-
-
assay at
30
-
-
assay at
37
-
-
assay at
37
-
-
assay at
37
-
C6EE50, -
assay at
37
-
Q01217, -
assay at
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
-
sheath and blade, coordinate expression of enzyme and PII-like protein GlnB during the life span
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
Escherichia coli (strain B / BL21-DE3)
Escherichia coli (strain B / BL21-DE3)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Pseudomonas aeruginosa (strain ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Streptococcus mutans serotype c (strain ATCC 700610 / UA159)
Synechococcus elongatus (strain PCC 7942)
Synechococcus elongatus (strain PCC 7942)
Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099)
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
30000
-
-
recombinant protein, SDS-PAGE
33000
-
-
mass spectrometry
36000
-
-
recombinant protein
36000
-
-
SDS-PAGE
93000
-
-
gel filtration
180000
-
-
sedimentation anaylsis
190000
-
-
gel filtration in presence of N-acetylglutamate
198000
-
-
calculated from amino acid sequence
230000
-
-
gel filtration
400000
-
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 52000, SDS-PAGE
?
-
x * 29000, SDS-PAGE
?
-
dimer or tetramer, 1 or 2 * 43000 + 1 or 2 * 53000, SDS-PAGE
dimer
-
2 * 30000, SDS-PAGE
dimer
-
crystalline structure
hexamer
-
6 * 30341, MALDI-TOF MS, 6 * 30344, calculated
hexamer
-
6 * 32000, calculated
hexamer
-
three homodimers are linked to form a ring-like hexamer
hexamer
Q9X2A4, -
three homodimers are linked to form a ring-like hexamer
hexamer
-
with an extra N-terminal-linked helix interlinking three dimers, the hexameric architecture is not essential for arginine inhibition but is functionally essential for physiologically relevant arginine control of NAGK
homodimer
C6EE50, -
-
tetramer
-
crystal structure
tetramer
Q01217, -
-
additional information
-
due to the capacity for self-association the enzyme can exist in different states of aggregation depending on the nature of the ligands and the concentration of phosphate buffer
additional information
-
enzyme hexamer forms a complex with PII signaling protein trimer; in vivo complex of enzyme with signal transduction protein PII
additional information
-
possible interaction of enzyme with PII signal proteins in vivo
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
side-chain modification
-
methylation of surface lysine residues
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
as complex with PII protein, NAGK binds Mg2+, ADP, arginine and N-acetylglutamate; hanging drop vapor diffusion method at room temperature, crystal structure of a complex formed between two homotrimers of PII and a single hexamer of enzyme bound to the metabolites N-acetylglutamate, ADP, ATP, and arginine
-
crystal structures of EcNAGK free from substrates or complexed with the product N-acetyl-L-glutamyl-5-phosphate (NAGP) and with sulfate are determined at 2 A resolution. Structures reveal a novel, very open NAGK conformation to which substrates associate and from which roducts dissociate. In this conformation, the C-domain, which hosts most of the nucleotide site, rotates 24-28 away from the N-domain, which hosts the acetylglutamate site, whereas the empty ATP site also exhibits some changes
C6EE50, -
in complex with MgADP-, N-acetyl-glutamte, AlF4-, with MgADP-, N-acetyl-glutamte, with ADp and SO42-
-
crystal structure of Maricaulis maris NAGS/K (mmNAGS/K) at 2.7 A resolution shows that it is a tetramer
-
without arginine
-
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
Q01217, -
hanging-drop vapor diffusion method
-
crystal structure of the complex between acetylglutamate kinase and PII of Synechococcus elongatus, at 2.75 A resolution
Q6V1L5, -
in complex with arginine
Q9X2A4, -
of the recombinant wild type protein, the selenomethionine substituted enzyme, the mutants and the methylated enzyme, cocrystallization with ATP, ADP, acetyl-CoA, CoA, N-acetyl-L-glutamate, adenylylimidodiphosphate, arginine
-
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
64
-
-
10 min, 75% loss of activity without stabilizer, about 40% loss of activity with 75 mM N-acetyl-L-glutamate, 10 min, 10% loss of activity with 1 mM L-arginine
70
-
-
30 min, complete inactivation
70
-
-
1 h, 100% residual activity
80
-
-
1 h, 80% residual activity
85
-
-
1 h, 60% residual activity
additional information
-
-
-
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
L-arginine protects against inactivating effects of high concentrations of urea
-
L-arginine protects against the inactivating effects of heat
-
urea, 4.0 M, complete loss of activity after 120 min
-
loss of activity on repeated freezing and thawing
-
loss of activity on repeated freezing and thawing
-
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-20C, stable over extended periods
-
4C, 0.1 M phosphate buffer, stable
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
affinity chromatography and gel filtration by binding to the PII protein, Ni-nitrilotriacetic acid agarose to purify the recombinant protein
-
His-Select nickel affinity gel column chromatography
-
of the recombinant protein
-
using Ni-NTA chromatography
-
using Ni-NTA chromatography
-
using Ni-NTA chromatography
Q01217, -
recombinant enzyme
-
of the recombinant protein
Q8P8J7
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expressed in Escherichia coli
-
expressed in Escherichia coli BL21(DE3) cells
-
expression in Escherichia coli
-
expressed in Escherichia coli
-
expressed in Escherichia coli as a His-tagged fusion protein
-
expressed in Escherichia coli as a His-tagged fusion protein
-
expressed in Escherichia coli as a His-tagged fusion protein
Q01217, -
expression in Escherichia coli
-
expressed in Escherichia coli BL21(DE3) cells
-
expression in Escherichia coli
-
expression in Escherichia coli, selenomethionine substituted enzyme, site directed mutagenesis
-
EXPRESSION
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
threefold repression of enzyme formation by arginine
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
E19R
-
Km and kcat (N-acetyl-L-glutamate) similar to wild-type, IC50 (L-arginine) approximately 55fold increased compared to wild-type
H268N
-
Km and kcat (N-acetyl-L-glutamate) similar to wild-type, IC50 (L-arginine) approximately 55fold increased compared to wild-type
H268N/H26E
-
Km and kcat (N-acetyl-L-glutamate) similar to wild-type, IC50 (L-arginine) 875fold increased compared to wild-type
H268N/H26E/E19R
-
Km and kcat (N-acetyl-L-glutamate) similar to wild-type, IC50 (L-arginine) 1960fold increased compared to wild-type
H26E
-
Km and kcat (N-acetyl-L-glutamate) similar to wild-type, IC50 (L-arginine) approximately 55fold increased compared to wild-type
DELTA2-15
-
Km (N-acetyl-L-glutamate) increased compared to wild-type, kcat decreased compared to wild-type, IC50 (L-arginine) 71fold increased compared to wild-type
DELTA2-19
-
Km (N-acetyl-L-glutamate) increased compared to wild-type, kcat decreased compared to wild-type, IC50 (L-arginine) 148fold increased compared to wild-type
DELTA2-29
-
Km (N-acetyl-L-glutamate) increased compared to wild-type, kcat decreased compared to wild-type, IC50 (L-arginine) 163fold increased compared to wild-type
DELTA25
-
Km (N-acetyl-L-glutamate) highly increased compared to wild-type, kcat highly decreased compared to wild-type, IC50 (L-arginine) 17fold increased compared to wild-type
DELTA8
-
Km (N-acetyl-L-glutamate) increased compared to wild-type, kcat highly decreased compared to wild-type, IC50 (L-arginine) 16fold increased compared to wild-type
E19A
-
Km (N-acetyl-L-glutamate) similar to wild-type, kcat similar to wild-type, IC50 (L-arginine) 40fold increased compared to wild-type
E19R
-
Km (N-acetyl-L-glutamate) similar to wild-type, kcat similar to wild-type, IC50 (L-arginine) 61fold increased compared to wild-type
E281A
-
Km (N-acetyl-L-glutamate) highly increased compared to wild-type, kcat similar to wild-type, IC50 (L-arginine) 21fold increased compared to wild-type
G287A
-
Km (N-acetyl-L-glutamate) similar to wild-type, kcat decreased compared to wild-type, IC50 (L-arginine) 37fold increased compared to wild-type
G287D
-
Km (N-acetyl-L-glutamate) similar to wild-type, kcat similar to wild-type, IC50 (L-arginine) 40fold increased compared to wild-type
H268A
-
Km (N-acetyl-L-glutamate) similar to wild-type, kcat similar to wild-type, IC50 (L-arginine) 39fold increased compared to wild-type
H268N
-
Km (N-acetyl-L-glutamate) similar to wild-type, kcat similar to wild-type, IC50 (L-arginine) 58fold increased compared to wild-type
H26A
-
Km (N-acetyl-L-glutamate) similar to wild-type, kcat similar to wild-type, IC50 (L-arginine) 54fold increased compared to wild-type
R209A
-
Km (N-acetyl-L-glutamate) similar to wild-type, kcat similar to wild-type, IC50 (L-arginine) 46fold increased compared to wild-type
R209K
-
Km (N-acetyl-L-glutamate) similar to wild-type, kcat similar to wild-type, IC50 (L-arginine) 37fold increased compared to wild-type
W23A
-
Km (N-acetyl-L-glutamate) similar to wild-type, kcat similar to wild-type, IC50 (L-arginine) 4.5fold increased compared to wild-type
D162E
-
about 0.1% of wild-type activity
G11A
C6EE50, -
G11A does not hamper recombinant enzyme expression or purification. Mutant shows a 10fold decrease in Vmax values and it selectively increases 8fold the Km for ATP, affecting much less (3fold increase) the apparent Km for N-acetyl-L-glutamate
K8R
-
substantial although diminished activity
N158K
-
substantial although diminished activity
R66K
-
substantial although diminished activity
I106M
-
to increase phasing power, three additional amino acids codons are mutated to methionine (I106M, I294M and L367M). Crystals from this mutant protein are diffracted to 2.7 A
I294M
-
to increase phasing power, three additional amino acids codons are mutated to methionine (I106M, I294M and L367M). Crystals from this mutant protein are diffracted to 2.7 A
L367M
-
to increase phasing power, three additional amino acids codons are mutated to methionine (I106M, I294M and L367M). Crystals from this mutant protein are diffracted to 2.7 A
E17A
-
site-directed mutagenesis, the mutant shows reduced Vmax, the mutation results in decreased affinity of NAGK for arginine
E17D
-
site-directed mutagenesis, the mutant shows reduced Vmax, the mutation results in decreased affinity of NAGK for arginine
E17Q
-
site-directed mutagenesis, the mutant shows reduced Vmax, the mutation results in decreased affinity of NAGK for arginine
E284D
-
site-directed mutagenesis, the mutant shows reduced Vmax and altered Km for the substrates
G290A
-
site-directed mutagenesis, the mutant shows reduced Vmax and altered Km for the substrates
H271N
-
site-directed mutagenesis, the mutant shows reduced Vmax and altered Km for the substrates
Q10A
-
site-directed mutagenesis, the mutant shows reduced Vmax and altered Km for the substrates
R24E
-
site-directed mutagenesis, the mutant shows reduced Vmax , the mutation results in increased affinity of NAGK for arginine
Y21A
-
site-directed mutagenesis, the mutant shows reduced Vmax and altered Km for the substrates
DELTA357-513
Q01217, -
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
R233A
-
mutant does not interact in vitro with PII protein of wild-type sequence in yeast two-hybrid analysis. Mutant interacts with PII variants containing I86N or I86T mutation
E186A/E387A
-
to improve resolution in crystallization
E416A/K417A
-
to improve resolution in crystallization
E94A/K95A
-
to improve resolution in crystallization
K26A/E27A
-
to improve resolution in crystallization
K279A/E280A
-
to improve resolution in crystallization
K419A
-
to improve resolution in crystallization
K213
-
site-directed mutagenesis, the mutant shows reduced Vmax and altered Km for the substrates
additional information
-
N-helix N-terminal deletions spanning 16 residues dissociate NAGK to active dimers, those of 20 residues decrease the apparent affinity for arginine, and complete N-helix deletion of 26 residues abolishes arginine inhibition, overview