Information on EC 2.3.1.17 - aspartate N-acetyltransferase and Organism(s) Homo sapiens

for references in articles please use BRENDA:EC2.3.1.17
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Homo sapiens


The taxonomic range for the selected organisms is: Homo sapiens

The enzyme appears in selected viruses and cellular organisms

EC NUMBER
COMMENTARY hide
2.3.1.17
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RECOMMENDED NAME
GeneOntology No.
aspartate N-acetyltransferase
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Acyl group transfer
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-
-
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Alanine, aspartate and glutamate metabolism
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Metabolic pathways
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SYSTEMATIC NAME
IUBMB Comments
acetyl-CoA:L-aspartate N-acetyltransferase
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CAS REGISTRY NUMBER
COMMENTARY hide
9029-99-6
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
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Canavan disease is a fatal, neurological disease that is caused by an interruption in the metabolism of a critical amino acid, N-acetyl-L-aspartic acid. Defects at multiple locations in the aspA gene that codes for aspartoacylase, EC 3.5.1.15, lead to mutant forms of this enzyme that are either not expressed or rapidly degraded, or have significantly impaired catalytic activity, resulting in N-acetyl-L-aspartic acid accumulation. A second gene knock-out in the Nat8l gene which codes for aspartate N-acetyltransferase, the enzyme that synthesizes N-acetyl-L-aspartic acid, reverses these adverse effects, leading to normal myelination and a decrease in Canavan disease symptoms
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 + 2,3-diaminosuccinate
CoA + ?
show the reaction diagram
-
-
-
-
?
acetyl-CoA + 3-methyl-L-aspartate
CoA + N-acetyl-3-methyl-L-aspartate
show the reaction diagram
-
-
-
-
?
acetyl-CoA + L-aspartate
CoA + N-acetyl-L-aspartate
show the reaction diagram
-
-
-
-
?
acetyl-CoA + L-aspartate
CoA + N-acetyl-L-aspartic acid
show the reaction diagram
acetyl-CoA + L-glutamate
CoA + N-acetyl-L-glutamate
show the reaction diagram
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reaction of EC 2.3.1.1
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-
?
acetyl-CoA + L-glutamate
CoA + N-acetyl-L-glutamic acid
show the reaction diagram
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less than 1% of the activity with L-aspartate
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-
?
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-aspartate
CoA + N-acetyl-L-aspartate
show the reaction diagram
-
-
-
-
?
acetyl-CoA + L-aspartate
CoA + N-acetyl-L-aspartic acid
show the reaction diagram
-
-
-
?
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
cholamido propane sulfonate
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cyclohexylmaltoside
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cymal5, high inhibition at CMC concentration
decylmaltoside
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DMSO
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20% inhibition at 40% v/v
dodecyl octaglycol
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dodecylmaltoside
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lauryldimethylamine-N-oxide
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complete inhibition at CMC concentration
n-decyl-N,N-dimethylamine-N-oxide
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high inhibition at CMC concentration
N-methyl-N-nonanoyl-beta-D-glucosylamine
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Mega-9, high inhibition at CMC concentration
n-nonyl-beta-D-glucopyranoside
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high inhibition at CMC concentration
octyl pentaglycol
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high inhibition at CMC concentration
octyl tetraglycol
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high inhibition at CMC concentration
octylglucoside
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high inhibition at CMC concentration
polymaleic anhydride C10
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polymaleic anhydride C12
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-
polymaleic anhydride C16
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high inhibition at CMC concentration
polymaleic anhydride C4
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high inhibition at CMC concentration
polymaleic anhydride C6
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complete inhibition at CMC concentration
polymaleic anhydride C8
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SDS
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complete inhibition at CMC concentration
sodium dodecanoyl sarcosine
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Triton X-100
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Tween 20
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additional information
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effect of different detergents on the enzyme activity: non-ionic detergents such as Triton X-100 are less disruptive to protein structures than ionic detergents such as SDS, detergents such as C12E8, Tween 20 and several maltosides caused minimal disruption of the enzyme, with greater than 50% residual activity after incubation with CMC levels of each of these detergents. In contrast, significant loss of activity is observed upon incubation with C8 detergents, cymal5, octylglucoside and some shorter chain polymaleic anhydride (pmal) detergents. Ionic detergent SDS and a zwitterionic detergent lauryldimethylamine-N-oxide cause nearly complete loss of catalytic activity
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Methamphetamine
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maximum increase in activity in SH-SY5Y cells is found at 1 microM methamphetamine at 24 h, increase in activity is about 2fold
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.92
2,3-diaminosuccinate
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pH 7.4, temperature not specified in the publication
0.36
3-methyl-L-aspartate
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pH 7.4, temperature not specified in the publication
0.001 - 0.4
acetyl-CoA
0.09 - 3.37
L-aspartate
8.6
L-glutamate
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pH 7.4, temperature not specified in the publication
additional information
additional information
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Michaelis-Menten kinetics
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.035
2,3-diaminosuccinate
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pH 7.4, temperature not specified in the publication
0.0018
3-methyl-L-aspartate
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pH 7.4, temperature not specified in the publication
0.0071
acetyl-CoA
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pH 7.4, temperature not specified in the publication
0.0071
L-aspartate
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pH 7.4, temperature not specified in the publication
0.023
L-glutamate
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pH 7.4, temperature not specified in the publication
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
70
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recombinant enzyme, pH 7.4, temperature not specified in the publication
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.4
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assay at
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 9.5
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the enzymatic activity decreases at pH values below pH 7.5. A fit of the Vmax/Km data to a model which assumes that the protonation of a single group leads to loss of activity results in a pK value of 6.8 for a group that must be ionized for the enzyme to remain catalytically active. By contrast, the Vmax profile does not show substantial changes across the pH range
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
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enzyme is present in both cytoplasm and mitochondria
Manually annotated by BRENDA team
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exclusively associated with the endoplasmic reticulum. the membrane region comprises alpha-helices and the catalytic site is in the cytosol. The membrane region, i.e. region 4, is necessary and sufficient to target isoform NAT8L to the endoplasmic reticulum
Manually annotated by BRENDA team
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membrane-associated, the enzyme contains a membrane anchor region
Manually annotated by BRENDA team
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enzyme is present in both cytoplasm and mitochondria
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
33900
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x * 56000-60000, recombinant MBP-His-tagged enzyme without membrane anchor, SDS-PAGE, x * 33900, recombinant His-tagged enzyme without membrane anchor, SDS-PAGE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
molecular modeling of the active site shows that only the amino acid aspartate, but not glutamate, can fit into the active site pocket
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purified recombinant His-tagged truncated enzyme, X-ray diffraction structure determination and analysis
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
recombinant functional and soluble dual His- and MBP-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and amylose affinity chromatography, MBP tag cleavage by 3C protease, method evaluation
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression HEK-293 cell
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expression in HEK-293T cell
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gene nat8l, sequence comparisons, subcloning of the genes for thioredoxin (TRX), glutathione S-transferase (GST) or maltose binding protein (MBP), followed by a linker region (21-68-amino acids) containing various cleavage site sequences, and then connected to the N-terminal of the nat8l gene, without the membrane anchor, recombinant functional and soluble expression of the dual affinity tagged enzyme as MBP-fusion protein in Escherichia coli strain BL21(DE3), method evaluation
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overexpressed both as native and as fusion protein with a His6 tag at the C- or the N-terminus in human embryonic kidney-293 cells expressing the large T-antigen of simian virus 40 (HEK-293T cells)
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C128A
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mutation in region 4, 94% of wild-type expression, 88% of wild-type activity
C139A
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mutation in region 4, 70% of wild-type expression, 126% of wild-type activity
D168A
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mutation in region 5, 46% of wild-type expression, no residual activity
D168E
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mutation in region 5, 63% of wild-type expression, 7% of wild-type activity
E101A
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mutation in region 3, 97% of wild-type expression, 42% of wild-type activity
E101D
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mutation in region 3, 61% of wild-type expression, 99% of wild-type activity
P142A
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mutation in region 4, 113% of wild-type expression, 116% of wild-type activity
R133A
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mutation in region 4, 78% of wild-type expression, 64% of wild-type activity
R133K
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mutation in region 4, 69% of wild-type expression, 89% of wild-type activity
R220A
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mutation in region 5, 101% of wild-type expression, 14% of wild-type activity
R220K
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mutation in region 5, 104% of wild-type expression, 25% of wild-type activity
R81A
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mutation in region 3, 57% of wild-type expression, 37% of wild-type activity
R81K
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mutation in region 3, 90% of wild-type expression, 82% of wild-type activity
S132F/R133F
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mutation in region 4, 92% of wild-type expression, 41% of wild-type activity
additional information
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transfection of truncated forms of isoform NAT8L into HEK-293T cells indicates that the 68 N-terminal residues, i.e. regions 1 and 2, have no importance for the catalytic activity and the subcellular localization of the enzyme. The membrane region, i.e. region 4, is necessary and sufficient to target isoform NAT8L to the endoplasmic reticulum