Information on EC 6.3.5.4 - asparagine synthase (glutamine-hydrolysing) and Organism(s) Homo sapiens and UniProt Accession P08243

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Homo sapiens
UNIPROT: P08243


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


The taxonomic range for the selected organisms is: Homo sapiens

EC NUMBER
COMMENTARY hide
6.3.5.4
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RECOMMENDED NAME
GeneOntology No.
asparagine synthase (glutamine-hydrolysing)
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
L-asparagine biosynthesis I
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aspartate and asparagine metabolism
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Alanine, aspartate and glutamate metabolism
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Metabolic pathways
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Biosynthesis of secondary metabolites
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SYSTEMATIC NAME
IUBMB Comments
L-aspartate:L-glutamine amido-ligase (AMP-forming)
The enzyme from Escherichia coli has two active sites [4] that are connected by an intramolecular ammonia tunnel [5,6]. The enzyme catalyses three distinct chemical reactions: glutamine hydrolysis to yield ammonia takes place in the N-terminal domain. The C-terminal active site mediates both the synthesis of a beta-aspartyl-AMP intermediate and its subsequent reaction with ammonia. The ammonia released is channeled to the other active site to yield asparagine [6].
CAS REGISTRY NUMBER
COMMENTARY hide
37318-72-2
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
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elevated expression of ASNS protein is associated with resistance to asparaginase therapy in childhood acute lymphoblastic leukemia and may be a predictive factor in drug sensitivity for certain solid tumors as well. Activation of the GCN2-eIF2-ATF4 signaling pathway, leading to increased ASNS expression, appears to be a component of solid tumor adaptation to nutrient deprivation and/or hypoxia, roles of the enzyme in fetal development, tissue differentiation, and tumor growth, overview. Possible correlation between ASNase sensitivity and the DNA methylation status of the ASNS gene
additional information
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human enzyme activity is highly regulated in response to cell stress, primarily by increased transcription from a single gene located on chromosome 7, ASNS transcription control by C/EBP-ATF response element within the promoter. Protein limitation or an imbalanced dietary amino acid composition activate the ASNS gene through the amino acid response, a process that is replicated in cell culture through limitation for any single essential amino acid
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
ATP + L-Asp + L-Gln
AMP + diphosphate + Asn + Glu
show the reaction diagram
ATP + L-Asp + NH3
AMP + diphosphate + Asn
show the reaction diagram
ATP + L-aspartate + L-glutamine
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
ATP + L-aspartate + L-glutamine + H2O
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
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?
additional information
?
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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
ATP + L-Asp + L-Gln
AMP + diphosphate + Asn + Glu
show the reaction diagram
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the basic region leucine zipper protein ATF5, a transcriptional activator, stimulates asparagine promoter/reporter gene transcription via the nutrient-sensing response unit
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?
ATP + L-aspartate + L-glutamine
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
ATP + L-aspartate + L-glutamine + H2O
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
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?
additional information
?
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upregulation of asparagine synthetase fails to avert cell cycle arrest induced by L-asparaginase in TEL/AML1-positive leukaemic cells
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
6-diazo-5-oxo-L-norleucine
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loss of Gln-dependent reactions, but no effect on ATP binding as measured during amminoa-dependent Asn synthesis
8-N3ATP
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loss of NH4+-dependent Asn synthesis, but no effect on the glutaminase activity
beta-asparaginyladenylate
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Gln
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0.4-2.0 mM, inhibits the ammonia-dependent production of Asn
mupirocin
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phosmidosine
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sulfoximine adenylate
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most potent inhibitor
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Phytohemagglutinin
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.38
aspartic acid
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pH 8, reaction with glutamine, C-terminally tagged recombinant enzyme
0.08 - 0.11
ATP
1.3
L-aspartic acid
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pH 8, reaction with NH3, C-terminally tagged recombinant enzyme
1.9
L-glutamine
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pH 8, C-terminally tagged recombinant enzyme
1.7
NH3
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pH 8, C-terminally tagged recombinant enzyme
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.6 - 1.7
ATP
1.7
glutamine
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pH 8, C-terminally tagged recombinant enzyme
1.3 - 1.7
L-aspartic acid
1.8
NH3
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pH 8, C-terminally tagged recombinant enzyme
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.000285
sulfoximine adenylate
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SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
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Manually annotated by BRENDA team
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Manually annotated by BRENDA team
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high expression in T-lineage and low expression in B-lineage
Manually annotated by BRENDA team
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higher expression than in lymphoblastic leukemia cells
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
64000
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SDS-PAGE
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
ATP protects from inactivation by UV irradiation in the presence of 8-N3ATP
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-80°C, recombinant C-terminally tagged enzyme is stable on prolonged storage
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
one-step immunoaffinity purification
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
C-terminally tagged enzyme, baculovirus-based expression system, the recombinant enzyme is correctly processed, exhibits high activity and is stable on prolonged storage at -80°C
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cloned into a 2 mü plasmid, pBS24.1GAS, suitable for replication in a Saccharomyces cerevisiae ciro strain AB116
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expression of AS-GFP fusion protein in MOLT-4 cells
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expression of several mutant enymes in Saccharomyces cerevisiae
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mutant enzyme in which the N-terminal Cys is replaced by Ala is expressed in Saccharomyces
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
human enzyme activity is highly regulated in response to cell stress, primarily by increased transcription from a single gene located on chromosome 7. Protein limitation or an imbalanced dietary amino acid composition activate the ASNS gene through the amino acid response, a process that is replicated in cell culture through limitation for any single essential amino acid. Endoplasmic reticulum stress also increases ASNS transcription through the PERK-eIF2-ATF4 arm of the unfolded protein response. Both the amino acid response and unfolded protein response lead to increased synthesis of ATF4, which binds to the C/EBP-ATF response element and induces ASNS transcription
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C1A
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altering Cys-1 to either Ala or Ser eliminated the Gln-dependent activity, while only minimally affecting the kinetic properties of the NH4+-dependent reaction
C1S
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altering Cys-1 to either Ala or Ser eliminated the Gln-dependent activity, while only minimally affecting the kinetic properties of the NH4+-dependent reaction
additional information
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the replacement of the N-terminal Cys by Ala results in the loss of the Gln-dependent Asn synthetase activity, while the NH4+-dependent activity remains unaffected
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
mass spectrometry-based procedure for the direct quantification of asparagine synthetase protein concentration in complex sample mixtures. Assay is able to distinguish samples from transformed cell lines that express the enzyme over a wide dynamic range of concentration. The method directly detects asparagine synthetase protein, use in blast samples from patients with acute lymphoblastic leukemia
medicine