Information on EC 2.3.1.1 - amino-acid N-acetyltransferase

Word Map on EC 2.3.1.1
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)


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

EC NUMBER
COMMENTARY hide
2.3.1.1
-
RECOMMENDED NAME
GeneOntology No.
amino-acid N-acetyltransferase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
acetyl-CoA + L-glutamate = CoA + N-acetyl-L-glutamate
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Acyl group transfer
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
L-arginine biosynthesis II (acetyl cycle)
-
-
L-arginine biosynthesis III (via N-acetyl-L-citrulline)
-
-
L-ornithine biosynthesis I
-
-
arginine metabolism
-
-
Arginine biosynthesis
-
-
Metabolic pathways
-
-
Biosynthesis of secondary metabolites
-
-
Biosynthesis of antibiotics
-
-
SYSTEMATIC NAME
IUBMB Comments
acetyl-CoA:L-glutamate N-acetyltransferase
Also acts with L-aspartate and, more slowly, with some other amino acids.
CAS REGISTRY NUMBER
COMMENTARY hide
9029-88-3
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
anamorph, Fusarium graminearum
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
gene Rv2747 or argA
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
suckling piglet
UniProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
-
enzyme deficiency results in elevated levels of plasma ammonia which is neurotoxic; enzyme deficiency results in elevated levels of plasma ammonia which is neurotoxic
metabolism
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
acetyl-CoA + DL-2-aminopimelate
CoA + 2-acetylaminoheptanedioate
show the reaction diagram
acetyl-CoA + glycine
CoA + acetylaminoacetate
show the reaction diagram
acetyl-CoA + L-2-aminoadipate
CoA + 2-acetylaminohexanedioate
show the reaction diagram
acetyl-CoA + L-aspartate
CoA + N-acetyl-L-aspartate
show the reaction diagram
-
3.0% of activity with L-glutamate
-
?
acetyl-CoA + L-glutamate
CoA + N-acetyl-L-glutamate
show the reaction diagram
acetyl-CoA + L-glutamate
N-acetyl-L-glutamate + CoA
show the reaction diagram
-
assay at pH 8.5, 30C, 5 min
-
-
?
acetyl-CoA + L-glutamate-gamma-hydroxamate
CoA + N-acetyl-L-glutamate-gamma-hydroxamate
show the reaction diagram
-
15.5% of activity with glutamate
-
?
acetyl-CoA + L-glutamine
?
show the reaction diagram
acetyl-CoA + L-glutamine
CoA + N-acetyl-L-glutamine
show the reaction diagram
acetyl-CoA + L-methionine
CoA + N-acetyl-L-methionine
show the reaction diagram
pitax is an acetyltransferase and weakly catalyses the acetylation of l-methionine and l-methionine sulfoxide. Pitax belongs to the GCN5-related N-acetyltransferase family and contains all four sequence motifs conserved among family members. The beta4-strand structure in one of these motifs is disrupted, which is believed to affect binding of the substrate that accepts the acetyl group from acetyl-CoA
-
-
-
acetyl-CoA + L-methionine sulfoxide
CoA + N-acetyl-L-methionine sulfoxide
show the reaction diagram
pitax is an acetyltransferase and weakly catalyses the acetylation of l-methionine and l-methionine sulfoxide. Pitax belongs to the GCN5-related N-acetyltransferase family and contains all four sequence motifs conserved among family members. The beta4-strand structure in one of these motifs is disrupted, which is believed to affect binding of the substrate that accepts the acetyl group from acetyl-CoA
-
-
-
benzoyl-CoA + L-glutamate
CoA + N-benzoyl-L-glutamate
show the reaction diagram
-
very low activity
-
?
butyryl-CoA + L-glutamate
CoA + N-butyryl-L-glutamate
show the reaction diagram
n-butyryl-CoA + L-glutamate
CoA + N-butyryl-L-glutamate
show the reaction diagram
-
-
1.5% of the activity with acetyl-CoA
-
?
n-propionyl-CoA + L-glutamate
CoA + N-propionyl-L-glutamate
show the reaction diagram
-
-
23% of the activity with acetyl-CoA
-
?
propionyl-CoA + L-glutamate
CoA + N-propionyl-L-glutamate
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
acetyl-CoA + L-glutamate
CoA + N-acetyl-L-glutamate
show the reaction diagram
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
acetyl-CoA
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1,3-Diaminopropane
-
10 mM, 88% inhibition in the presence of 0.5 mM N-acetyl-L-glutamate
3-methylcrotonyl-CoA
-
about 45% residual activity at 2.5 mM
5,5'-dithiobis(2-nitrobenzoate)
-
strong inhibition
acetyl-CoA
AgNO3
-
0.1 mM, 70-90% inhibition
arginine
BaCl2
-
1 mM, 30-50% inhibition
Butyryl-CoA
-
about 30% residual activity at 2.5 mM
CaCl2
-
1 mM, 30-50% inhibition
cadaverine
-
10 mM, 80% inhibition in the presence of 0.5 mM N-acetyl-L-glutamate
Cd2+
-
0.1 mM, 94% inhibition
CoA
-
50% activity reduction at 2.5 mM
CoCl2
-
0.1 mM, 30-50% inhibition
coenzyme A
EDTA
-
weak inhibition
FeCl3
-
0.1 mM, 30-50% inhibition
FeSO4
-
0.1 mM, 30-50% inhibition
glutaryl-CoA
-
about 50% residual activity at 2.5 mM
High ionic strength
-
isobutylmethylxanthine
-
and other oxypurines containing a 2,6-dione group interfere with the binding of glutamate to the active site of N-acetylglutamate synthetase, thereby decreasing synthesis of N-acetylglutamate, resulting in reduction of citrulline and urea synthesis. Isobutylmethylxanthine significantly increases the apparent Km for glutamate and decreases velocity of N-acetylglutamate synthetase, with little effect on carbamoylphosphate synthase-1. Inhibition is reversed by supplementation with N-carbamylglutamate
isobutyryl-CoA
-
about 45% residual activity at 2.5 mM
isovaleryl-CoA
-
about 50% residual activity at 2.5 mM
L-alpha-Acetoxylglutamate
-
2 mM, 17% inhibition
L-arginine
L-citrulline
-
10 mM, 75% inhibition
L-glutamate
-
-
L-glutamine
-
substrate inhibition
L-Indospicine
-
0.2 mM; 50% inhibition
methylmalonyl-CoA
-
about 40% residual activity at 2.5 mM
MgCl2
-
1 mM, 30-50% inhibition
MnCl2
-
0.1 mM, 30-50% inhibition
N-acetyl-D-glutamate
-
2 mM, 30% inhibition
N-acetyl-DL-alpha-aminoadipate
-
2 mM, 78% inhibition
N-acetyl-L-aspartate
-
2 mM, 25% inhibition
N-acetyl-L-glutamate
N-acetyl-L-glutamine
-
2 mM, 46% inhibition
N-acetylglutamate
N-benzoyl-L-glutamate
-
2 mM, 29% inhibition
N-Butyryl-L-glutamate
-
2 mM, 19% inhibition
N-carbamoyl-L-glutamate
-
2 mM, 31% inhibition
N-ethylmaleimide
-
-
N-propionyl-L-glutamate
-
2 mM, 63% inhibition
NaCl
-
200 mM NaCl inhibits the activity by about 22%
Ni(NO3)2
-
0.1 mM, 30-50% inhibition
O-(L-Norvalyl-5)-isourea
-
0.02 mM, 50% inhibition
oxaloacetate
-
1 mM, 68% inhibition
p-chloromercuribenzoate
-
-
p-hydroxymercuribenzoate
Pb(NO3)2
-
0.1 mM, 70-90% inhibition
Polyamines
-
-
-
potassium phosphate
-
-
propionyl-CoA
putrescine
-
10 mM, 74% inhibition in the presence of 0.5 mM N-acetyl-L-glutamate
Sodium acetate
-
100 mM NaCl inhibits the activity by about 22%
spermidine
-
1 mM, 78% inhibition in the presence of 0.5 mM N-acetyl-L-glutamate
spermine
-
1 mM, 88% inhibition in the presence of 0.5 mM N-acetyl-L-glutamate
succinate
-
2 mM, 21% inhibition
succinyl-CoA
-
about 60% residual activity at 2.5 mM
uric acid
-
significantly increases the apparent Km for glutamate and decreases velocity of N-acetylglutamate synthetase, with little effect on carbamoylphosphate synthase-1. Inhibition is reversed by supplementation with N-carbamylglutamate
xanthine
-
significantly increases the apparent Km for glutamate and decreases velocity of N-acetylglutamate synthetase, with little effect on carbamoylphosphate synthase-1. Inhibition is reversed by supplementation with N-carbamylglutamate
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
arginine
Cationic polypeptides
-
4fold activation
-
culpein
-
-
-
L-arginine
L-Argininic acid
-
activates to a lesser extent than L-arginine
Polyarginine
-
-
polylysine
-
-
salmin
-
-
-
Triton X-100
-
0.1%, 4fold activation
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.05 - 8.1
acetyl-CoA
1.6 - 8.1
glutamate
1 - 557
L-glutamate
280
L-glutamine
-
-
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.22
L-glutamate
Mycobacterium tuberculosis
-
-
0.78
L-glutamine
Mycobacterium tuberculosis
-
-
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.015 - 15.1
acetyl-CoA
0.07
acetyl-glutamate
-
-
1.3
Butyryl-CoA
-
at pH 8.5 and 30C
1.9
isovaleryl-CoA
-
at pH 8.5 and 30C
60 - 152
L-glutamate
30
L-glutamine
-
-
0.49
N-acetyl-L-glutamate
-
-
0.71
propionyl-CoA
-
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2.5
CoA
Escherichia coli
-
pH and temperature not specified in the publication
0.36
isobutylmethylxanthine
Rattus norvegicus
-
pH 7.4, 37C
0.19 - 2
L-arginine
25
N-acetyl-L-glutamate
Escherichia coli
-
pH and temperature not specified in the publication
0.24
uric acid
Rattus norvegicus
-
pH 7.4, 37C
0.17
xanthine
Rattus norvegicus
-
pH 7.4, 37C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.0000135
-
activity in liver of fetus, activity increases strongly after eating external diets until week 10 after birth
0.0000136
-
activity in intestine of fetus
0.000014
-
-
0.0000203
-
activity in intestine at day 3 after birth
0.0000455
-
activity in intestine of adult
0.0000643
-
activity in intestine at day 1 after birth
0.00013
0.000268
0.002
-
pH 8.0, 70C, activity in cell culture in growth medium with 0.2% yeast extract; pH 8.0, 70C, activity in cell culture in minimal growth medium with 20 mM glucose and 5 mM NH4+
0.00286
-
-
0.053
-
purified recombinant His-tagged enzyme, in absence of L-arginine
0.078
-
mutant enzyme Y485F, at pH 8.5 and 30C
0.081
-
purified recombinant His-tagged enzyme, in presence of L-arginine
0.098
-
wild-type
0.136
-
wild-type + 1 mM arginine
0.139
-
purified recombinant His-tagged enzyme in 10fold dilution, in absence of L-arginine
0.154
-
mutant enzyme N479A, at pH 8.5 and 30C
0.163
-
purified recombinant His-tagged enzyme in 10fold dilution, in presence of L-arginine
0.325
-
purified recombinant His-tagged enzyme in 50fold dilution, in absence of L-arginine
0.346
mature recombinant enzyme in the absence of arginine
0.359
-
purified recombinant His-tagged enzyme in 50fold dilution, in presence of L-arginine
0.438
-
wild-type
0.528
-
wild-type + 1 mM arginine
0.572
-
wild-type + 1mM arginine
0.803
mature recombinant enzyme in the presence of 1 mM arginine
0.857
-
mutant enzyme Y441F, at pH 8.5 and 30C
1.05
-
wild type enzyme, at pH 8.5 and 30C
1.84
-
wild-type + 1mM arginine
3.19
-
mutant G362S + 1 mM arginine
3.22
-
mutant G362S
3.39
-
mutant G286P + 1 mM arginine; mutant G286P + 1mM arginine
3.51
-
mutant G286P
4.31
-
mutant F35C + 1 mM arginine; mutant F35C + 1mM arginine
4.4
residual activity at inhibition by L-arginine
4.97
mutant enzyme S387A, at pH 8.5 and 30C
5.475
-
wild-type + 1 mM arginine
5.85
-
wild-type
6.81
wild type enzyme, at pH 8.5 and 30C
9.11
-
wild-type
9.66
-
mutant F35C
10.64
-
wild-type
12.22
-
mutant F121C
12.27
-
wild-type
12.57
-
mutant G360P + 1 mM arginine; mutant G360P + 1mM arginine
13
-
mutant G360P
13.08
-
mutant F121C + 1 mM arginine
15.28
-
mutant E354A + 1 mM arginine
15.47
-
mutant E354A
15.89
-
wild-type + 1 mM arginine
20.88
-
wild-type
24
-
mutant delE283delQ284
25.56
-
wild-type + 1 mM arginine
26.68
-
mutant E280A + 1 mM arginine; mutant E280A + 1mM arginine
26.93
-
mutant E280A
42.55
-
mutant G288S + 1 mM arginine; mutant G288S + 1mM arginine
43.6
-
mutant ins284Ains285A
46.04
-
mutant G288S
48.8
-
mutant ins284A
62
-
wild-type
79.4
-
wild-type enzyme
92.4
-
in the presence of 1 mM L-arginine
111.7
-
wild-type
114.9
-
mutant delQ284
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
8 - 9.5
-
38% of maximal activity at pH 8.0., 25% of maximal activity at pH 9.5
8.5 - 10
-
pH 8.5: about 65% of maximal activity, pH 10.0: about 80% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
-
assay at
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
mucosa
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Bacillus halodurans (strain ATCC BAA-125 / DSM 18197 / FERM 7344 / JCM 9153 / C-125)
Enterococcus faecalis (strain ATCC 700802 / V583)
Maricaulis maris (strain MCS10)
Maricaulis maris (strain MCS10)
Maricaulis maris (strain MCS10)
Maricaulis maris (strain MCS10)
Neisseria gonorrhoeae (strain ATCC 700825 / FA 1090)
Neisseria gonorrhoeae (strain ATCC 700825 / FA 1090)
Neisseria gonorrhoeae (strain ATCC 700825 / FA 1090)
Neisseria gonorrhoeae (strain ATCC 700825 / FA 1090)
Neisseria gonorrhoeae (strain ATCC 700825 / FA 1090)
Streptococcus pneumoniae serotype 4 (strain ATCC BAA-334 / TIGR4)
Vibrio parahaemolyticus serotype O3:K6 (strain RIMD 2210633)
Xanthomonas campestris pv. campestris (strain 8004)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30000 - 300000
-
gel filtration, mixture of multiple forms
37400
catalytic N-acetyltransferase domain, gel filtration; recombinant dimeric N-acetyltransferase domain, gel filtration
38800
catalytic N-acetyltransferase domain, calculated from amino acid sequence
49070
-
SDS-PAGE
160000
-
gel filtration, sucrose density gradient centrifugation
190000
-
gel filtration
202400
-
calculated from amino acid sequence
220100
-
gel filtration
330000
gel filtration
additional information
-
-
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hexamer
homodimer
x-ray crystallography
homotetramer
tetramer
trimer
-
3 * 57000, SDS-PAGE
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
proteolytic modification
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
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; 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, 18C, X-ray diffraction structure determination and analysis at 2.1 A resolution, molecular replacement
-
development of a structural model of human enzyme that is fully consistent with the functional effects of the 14 missense mutations that have been identified in N-acetylglutamate synthase-deficient patients
-
N-acetyl-L-glutamate synthase/kinase in complex with L-arginine, sitting drop vapor diffusion method, using 100 mM sodium cacodylate trihydrate, pH 6.2, 25% (w/v) polypropylene glycol P400 and 200 mM magnesium chloride
to 2.7 A resolution. Enzyme is a tetramer. Each subunit has an amino acid kinase domain, which is likely responsible for N-acetylglutamate kinase activity and has a putative arginine binding site, and an N-acetyltransferase domain that contains the putative N-acetylglutamate synthase active site. The angle of rotation between amino acid kinase and N-acetyltransferase domains varies among crystal forms and subunits within the tetramer. A rotation of 26 is sufficient to close the predicted AcCoA binding site, thus reducing enzymatic activity
hanging drop method
-
purified recombinant enzyme, hanging drop vapour diffusion method, mixing of 0.001 ml protein solution containing 13 mg/ml protein with 0.001 ml of reservoir solution containing 0.1 M Tris-HCl, pH 6.5, 1.6 M ammonium sulfate, and 0.1% w/v sodium azide, 18C, 1-2 days, cryoprotection in 0.1 M Tris-HCl, pH 6.5, 1.6 M ammonium sulfate, 30% v/v glycerol and 0.1% w/v sodium azide, X-ray diffraction structure determination and analysis at 2.25 A resolution
sitting drop and hanging-drop vapor diffusion method, crystals belong to the hexagonal space group P6(2)22, with unit-cell parameters a = b = 134.60, c = 192.11 A, and diffract to about 3.0 A resolution
-
enzyme bound to N-acetyl-L-glutamate, sitting drop vapor diffusion method, using 0.2 M Li2SO4, 0.1 M Tris pH 6.5, 25% (w/v) PEG 3350; purified recombinant N-acetyltransferase domain of N-acetyl-L-glutamate synthase/kinase, with and without a His-tag, complexed with N-acetyl-L-glutamate, sitting-drop vapor-diffusion method, 0.002 ml of protein in 50 mM Tris-HCl, pH 7.4, 50 mM NaCl, 10% glycerol, 5 mM 2-mercaptoethanol, 1 mM EDTA, 10 mM CoA, and 10 mM N-acetyl-L-glutamate, is mixed with 0.2 M Li2SO4, 0.1 M Tris, pH 6.5, 25% PEG 3350 for the His-tagged enzyme, and with 0.2 M Li2SO4, 0.1 M Tris, pH 8.5, 25% PEG 3350 for the detagged enzyme, X-ray diffraction structure determination and analysis at 1.7 A and 1.4 A resolution, respectively
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7
-
highest stability
486061, 486063
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
38
-
strong decrease in activity after 5 min in the absence of arginine, presence of 1 mM arginine stabilizes
60
-
5 min, complete loss of activity
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
glycerol stabilizes
-
L-arginine or N-acetyl-L-glutamate stabilizes the high molecular weight form of 300000 Da
-
Triton X-100 stabilizes
-
Triton X-100, 0.1%, stabilizes liver enzyme
-
use of silicone-treated glassware or plastic tubes, e.g. polyethylene, polycarbonate or polypropylene stabilizes the enzyme
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, stable for 2 months
-
-80C, 100 mM phosphate buffer, pH 7.5, 6 months, 50% loss of activity
-
4C, 100 mM potassium phosphate buffer, pH 7.5, 24 h, 10% loss of activity
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ammonium sulfate, DEAE Biogel, Sephacryl S-200, aminoacetyl biogel A, affi-gel blue, sucrose gradient, isoelectric focusing
-
ammonium sulfate, hydroxyapatite
-
ammonium sulfate, hydroxyapatite, 10fold purification
-
ammonium sulfate, hydroxyapatite, DEAE-cellulose, Sephacryl-300
-
ammonium sulfate, hydroxyapatite, DEAE-cellulose, Sephadex G-100
-
DEAE-cellulose, hydroxylapatite
-
gel filtration
-
HisTrap nickel affinity column chromatography
-
HiTrap column chromatography; recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and cation exchange chromatography, the His-tag is cleaved by thrombin
nickel affinity and DEAE columns
-
nickel affinity and Histrap column chromatography; 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
-
nickel affinity column chromatography and DEAE column chromatography
recombinant enzyme, Ni2+-affinity column
recombinant His-tagged enzyme from Escherichia coli by nickel affinity chromatography, ammonium sulfate fractionation, gel filtration, and ultrafiltration
-
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
recombinant His-tagged wild-type enzyme by nickel affinity chromatography
-
using Ni-NTA chromatography
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
cloned in Escherichia coli as a His-tagged fusion protein
cloning of N-acetylglutamate synthase gene argA and a mutant gene fbr-argA responsible for arginine feedback resistance
coexpression of N-acetylglutamate synthase gene ARG2 together with ARG5 and ARG6 coding for acetylglutamate kinase and acetylglutamyl-phosphate reductase in Saccharomyces cerevisiae
-
expressed in an Escherichia coli JM109 DELTAargA mutant; gene cg3035, genomic organisation, gene cg3035 has an operon structure together with gene cg3034, expression of gene cg3035 complements an argA-deficient Escherichia coli mutant JM109 strain, recombinant overexpression in Corynebacterium glutamicum strain ATTC 13032
CAF20762, Q8NM40
expressed in Escherichia coli BL21 AI cells
-
expressed in Escherichia coli BL21(DE3) cells
expressed in Escherichia coli BL21(DE3) cells; recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)
expressed in Escherichia coli BL21(DE3) cells; recombinant expression of His-tagged wild-type and mutant enzymes, and isolated catalytic N-acetyltransferase domain in Escherichia coli strain BL21(DE3)
-
expressed in Escherichia coli C41(DE3) cells
-
expression in Escherichia coli
expression in Escherichia coli BL21
gene PA1377, cloning in Escherichia coli strain JM109, expression in Escherichia coli strain BL21(DE3)
gene Rv2747 or argA, expression of the His-tagged enzyme in Escherichia coli
-
gene structure, His-tagged wild-type and mutant enzymes in enzyme-deficient Escherichia coli strain NK 5992; mutent enzymes from patients with NAGS missense mutations are overexpressed in Escherichia coli NK5992. All mutated proteins show severe decrease in enzyme activity providing evidence for the disease-causing nature of the mutations
-
overexpression of His-tagged enzyme in Escherichia coli strain BL21(DE3), functional complementation of an enzyme-deficient Escherichia coli strain argA-
-
overexpression of His-tagged enzyme in Escherichia coli strain BL21(DE3), functional complementation of an enzyme-deficient Escherichia coli strain argA-, overexpression of the preenzyme in insect cells via baculovirus infection system, 2 processed forms with different N-terminal truncations are produced
-
phylogenetic analysis
-
transfection to Arabidopsis thaliana
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
3 h after exposure to low oxygen concentration decrease in transcript levels, 3 h after exposure to air rapid decrease of transcript levels
high expression at red fruit stage, first 3 h after exposure to low oxygen concentration higher transcript levels, 3 h after exposure to ethylene higher transcript levels
intestinal mRNA levels are lower in 7- to 28-day-old than in 1-day-old pigs
no repression by arginine
the enzyme is expressed between 24 and 48 h post fertilization
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
G287S
arginine feedback resistant mutant enzyme
H15Y
arginine feedback resistant mutant enzyme
Q432R
arginine feedback resistant mutant enzyme
R58H
arginine feedback resistant mutant enzyme
S54N
arginine feedback resistant mutant enzyme
Y19C
arginine feedback resistant mutant enzyme
A279T
-
mutations that causes NAGS deficiency, late onset of disease
E433D
-
mutations that causes NAGS deficiency, late onset of disease
L312P
-
mutations that causes NAGS deficiency, late onset of disease
L442V
-
mutations that causes NAGS deficiency, late onset of disease
N479A
-
site-directed mutagenesis, an active site mutant with reduced activity compared to the wild-type enzyme; the mutant shows reduced activity compared to the wild type enzyme
R509Q
-
mutations that causes NAGS deficiency, late onset of disease
T431I
-
mutations that causes NAGS deficiency, late onset of disease
V173E
-
mutations that causes NAGS deficiency, late onset of disease
V350I
-
mutations that causes NAGS deficiency, late onset of disease
W324X
-
mutations that causes NAGS deficiency, neonatal onset of disease
Y441F
-
site-directed mutagenesis, an active site mutant with reduced activity compared to the wild-type enzyme; the mutant shows reduced activity compared to the wild type enzyme
Y485F
-
site-directed mutagenesis, an active site mutant with reduced activity compared to the wild-type enzyme; the mutant shows reduced activity compared to the wild type enzyme
K356H
inactive
N391Q
inactive
R386K
inactive
S387A
the mutant shows reduced activity compared to the wild type enzyme
Y397F
inactive
E354A
-
specific activity (micromol/min/mg): 15.47, 15.28 (+ 1mM arginine), arginine activation is abolished; specific activity (micromol/min/mg): 15.47, 15.28 (+1mM arginine), arginine activation is abolished
F121C
-
specific activity (micromol/min/mg): 12.22, 13.08 (+ 1mM arginine), arginine activation is abolished; specific activity (micromol/min/mg): 12.22, 13.08 (+1mM arginine), arginine activation is abolished
G360P
-
specific activity (micromol/min/mg): 13.00, 12.57 (+ 1mM arginine), arginine activation is abolished; specific activity (micromol/min/mg): 13.00, 12.57 (+1mM arginine), arginine activation is abolished
G362S
-
specific activity (micromol/min/mg): 3.22, 3.19 (+ 1mM arginine), arginine activation is abolished; specific activity (micromol/min/mg): 3.22, 3.19 (+1mM arginine), arginine activation is abolished
D336N
-
point mutation
P427S
-
point mutation
V312I
-
point mutation
delE283delQ284
-
decrease of enzyme activity, induced substrate inhibition by acetyl-CoA
delQ284
-
increase of enzyme activity, triggered acetyl-CoA inhibition
E269A
-
mutation affects the putative arginine site
E352A
-
mutation affects the GCN5-related N-acetyltransferase domain
E352D
-
mutation affects the GCN5-related N-acetyltransferase domain
G146C
-
corresponding to clinical mutation
G275A
-
mutation affects the putative arginine site
G339A
-
mutation affects the GCN5-related N-acetyltransferase domain
G368A
-
mutation affects the GCN5-related N-acetyltransferase domain
ins284A
-
lower enzyme activity, decreased inhibition by arginine
ins284Ains285A
-
lower enzyme acitivity, decreased inhibition by arginine
K199A
-
mutation affects the putative arginine site
L353V
-
corresponding to clinical mutation
V358A
-
mutation affects the GCN5-related N-acetyltransferase domain
Y14A
-
mutation affects the putative arginine site
E280A
-
specific activity (micromol/min/mg): 26.93, 26.68 (+ 1mM arginine), arginine inhibition is abolished; specific activity (micromol/min/mg): 26.93, 26.68 (+1mM arginine), arginine inhibition is abolished
F35C
-
specific activity (micromol/min/mg): 9.66, 4.31 (+ 1mM arginine), only partial inhibition by arginine; specific activity (micromol/min/mg): 9.66, 4.31 (+1mM arginine), only partial inhibition by arginine
G286P
-
specific activity (micromol/min/mg): 3.51, 3.39 (+ 1mM arginine), no inhibition by arginine; specific activity (micromol/min/mg): 3.51, 3.39 (+1mM arginine), no inhibition by arginine
G288S
-
specific activity (micromol/min/mg): 46.04, 42.55 (+ 1mM arginine), no inhibition by arginine; specific activity (micromol/min/mg): 46.04, 42.55 (+1mM arginine), no inhibition by arginine
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
-
due to the close phylogenetic relationship and similar biochemical properties of xanthomonad NAGS-K and mammalian NAGS the enzyme from Xanthomonas campestris could become a good model for mammalian NAGS in structural, biochemical and biophysical studies
medicine
additional information
-
N-acetylglutamate synthase is a molecular marker for evolution of tetrapods
Show AA Sequence (3785 entries)
Please use the Sequence Search for a certain query.