Information on EC 6.3.5.4 - Asparagine synthase (glutamine-hydrolysing)

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

EC NUMBER
COMMENTARY
6.3.5.4
-
RECOMMENDED NAME
GeneOntology No.
Asparagine synthase (glutamine-hydrolysing)
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ATP + L-aspartate + L-glutamine + H2O = AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
mechanism
-
ATP + L-aspartate + L-glutamine + H2O = AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
uni-uni-bi-ter ping-pong mechanism without abortive complexes. Gln binds first, followed by Glu release, and Asp and ATP bind in order followed by ordered release of diphosphate, AMP and Asn. In the presence of 0.5-2.0 mM excess Mg2+ over ATP the binding of substrates after the release of Glu is in a rapid equilibrium system
-
ATP + L-aspartate + L-glutamine + H2O = AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
overall ping-pong mechanism
-
-
-
ATP + L-aspartate + L-glutamine + H2O = AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
NH4+ is bound to the enzyme followed by MgATP causing Asn release
-
ATP + L-aspartate + L-glutamine + H2O = AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
Arg325 is involved in stabilization of a pentacovalent intermediate leading to the formation of beta-aspartyl-AMP
-
ATP + L-aspartate + L-glutamine + H2O = AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
a model for the role of the catalytic triad in transferring nitrogen from Gln to Asp. An alternative catalytic mechanism is proposed, which obviates the participation of a histidine residue in the reaction
-
ATP + L-aspartate + L-glutamine + H2O = AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
hybrid uni uni bi ter ping pong Theorell-Chance mechanism where the glutaminase reaction occurs first and Asp binds to the enzyme before ATP in the sequential segment. Diphosphate is the first product released in the Theorell-Chance reaction, which is followed by the ordered release of AMP and Asn
-
ATP + L-aspartate + L-glutamine + H2O = AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
ping-pong reaction mechanism. Glutamine is the first substrate to bind to the enzyme and Asn is the last product released
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Acid amide hydrolysis
-
-
carboxylic
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Alanine, aspartate and glutamate metabolism
-
-
aspartate and asparagine metabolism
-
-
Biosynthesis of secondary metabolites
-
-
L-asparagine biosynthesis I
-
-
Metabolic pathways
-
-
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].
SYNONYMS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
AS
Q84LA5
-
AS-B
-
-
-
-
AS1
Q6HA26
-
ASNase
-
-
AsnB
-
-
-
-
ASNS
Q2L9S5
-
ASNS
P08243
-
ASNS
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
-
AsnS1
B4FFJ0
-
AsnS2
B5U8J7
-
AsnS3
B5U8J8
-
AsnS4
B5U8J9
-
asparagine amidotransferase
A9XS73
-
Asparagine synthetase
Q2L9S5
-
Asparagine synthetase
-
-
Asparagine synthetase
-
-
Asparagine synthetase
P08243
-
Asparagine synthetase
-
-
Asparagine synthetase
-
-
Asparagine synthetase
A9XS73
-
Asparagine synthetase
D9IXC8, E7EAQ2
-
Asparagine synthetase
Q6HA26
-
Asparagine synthetase
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
-
Asparagine synthetase (glutamine hydrolyzing)
-
-
-
-
Asparagine synthetase (glutamine)
-
-
-
-
Asparagine synthetase (glutamine-hydrolysing)
-
-
-
-
asparagine synthetase 1
-
-
asparagine synthetase 2
-
-
Asparagine synthetase B
-
-
-
-
Asparagine synthetase B
-
-
asparagine synthetase, glutamine-dependent
-
-
glutamine-dependent amidotransferase
-
-
Glutamine-dependent asparagine synthetase
-
-
-
-
HvAS1 protein
Q93XP9
Hordeum vulgare
HvAS2 protein
Q84LA5
Hordeum vulgare
L-asparaginase
-
-
L-Asparagine synthetase
-
-
-
-
PVAS1 protein
Q9SM55
Phaseolus vulgaris
Ste10
Q8KN11
-
Synthetase, Asn (glutamine)
-
-
-
-
TS11 cell cycle control protein
-
-
-
-
type -II asparagine synthetase
-
-
type I asparagine synthetase
Q9SM55
-
type II asparagine synthetase
-
-
CAS REGISTRY NUMBER
COMMENTARY
37318-72-2
-
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain 168, three glutamine-dependent AsnB-type asparagine snythetase genes: asnB, asnH and asnO, formerly yisO
-
-
Manually annotated by BRENDA team
Bacillus subtilis 168
strain 168, three glutamine-dependent AsnB-type asparagine snythetase genes: asnB, asnH and asnO, formerly yisO
-
-
Manually annotated by BRENDA team
ox
-
-
Manually annotated by BRENDA team
; gene CaAS1
-
-
Manually annotated by BRENDA team
the organism contains both enzymes EC 6.3.1.1. and EC 6.3.4.5. The gene asnA codes for NH4+-dependent Asn synthetases, EC 6.3.1.1, and the gene asnB codes for Gln-dependent Asn synthetase, EC 6.3.5.4
-
-
Manually annotated by BRENDA team
Asn synthetase B
-
-
Manually annotated by BRENDA team
Asn synthetase B; wild-type enzyme and mutant N74S
-
-
Manually annotated by BRENDA team
wild-type and mutant enzymes C1A, C1S, H29A , H80A, D33N, D33E, and A104H
-
-
Manually annotated by BRENDA team
wild-type and mutant enzymes C99A, C168A, C386A, C423A, C436A, C514A, C523A
-
-
Manually annotated by BRENDA team
wild-type and mutant enzymes R30A, R30K, N74A, N74Q, N79A
-
-
Manually annotated by BRENDA team
wild-type and mutant enzymes, E317A, E317Q, T318A, Y319A, Y319F, D320A, V321A, T322A, T322S, T322V, T322Y, T323A, T323I, T323L, T323S, T323V, R325A, R325K, T328S
-
-
Manually annotated by BRENDA team
wild-type and mutant N74A
-
-
Manually annotated by BRENDA team
cv. Corsoy
-
-
Manually annotated by BRENDA team
isoform SAS3
UniProt
Manually annotated by BRENDA team
isoforms SAS1 and SAS2
-
-
Manually annotated by BRENDA team
isoforms SAS1, SAS2
-
-
Manually annotated by BRENDA team
Merr. v. DP3588
-
-
Manually annotated by BRENDA team
cultivar HA-89
-
-
Manually annotated by BRENDA team
HAS1.1
SwissProt
Manually annotated by BRENDA team
patients with acute lymphoblastic leukemia
SwissProt
Manually annotated by BRENDA team
cv. Alexis, HvAS1, two AS genes: HvAS1 and HvAS2
SwissProt
Manually annotated by BRENDA team
cv. Alexis, HvAS2, two AS genes: HvAS1 and HvAS2
SwissProt
Manually annotated by BRENDA team
v. Victoria
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
cv. Sasanishiki
-
-
Manually annotated by BRENDA team
cv. Great Northern
SwissProt
Manually annotated by BRENDA team
cv. Negro Jamapa, gene NAS2
UniProt
Manually annotated by BRENDA team
AS1; gene PpAS1
UniProt
Manually annotated by BRENDA team
AS2; gene PpAS2
UniProt
Manually annotated by BRENDA team
v. Little Marvel, dark-grown
-
-
Manually annotated by BRENDA team
2 distinct Gln-dependent Asn synthetases
-
-
Manually annotated by BRENDA team
wild-type and auxotroph mutants
-
-
Manually annotated by BRENDA team
isoforms StAs1 and StAs2
-
-
Manually annotated by BRENDA team
strain 139
UniProt
Manually annotated by BRENDA team
enzyme form TaSN1; enzyme form TaSN2
-
-
Manually annotated by BRENDA team
isoform AsnS1; isozyme AsnS1; isozyme AsnS1
-
Manually annotated by BRENDA team
isoform AsnS2; isozyme AsnS2; isozyme AsnS2
-
Manually annotated by BRENDA team
isoform AsnS3; isozyme AsnS3; isozyme AsnS3
-
Manually annotated by BRENDA team
isoform AsnS4; isozyme AsnS4; isozyme AsnS4
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
evolution
-
CaAS1 and its related plant proteins all contain an AsnB domain and an ASN synthetase domain
evolution
D9IXC8, E7EAQ2
genes PpAS1 and PpAS2 encode both class II asparagine synthetases suggesting an ancient origin of the genes in plants. Amino acid residues essential for aspartate, AMP, and glutamine binding are conserved
evolution
D9IXC8, E7EAQ2
genes PpAS1 and PpAS2 encode both class II asparagine synthetases suggesting an ancient origin of the genes in plants. Amino acid residues essential for aspartate, AMP, and glutamine binding are conserved, except valine268, included in the AMP binding site, which has been replaced by an isoleucine in the AS2 sequence
malfunction
-
salt tolerance of Arabidopsis knockout mutant with T-DNA insertion in ASN2 gene encoding asparagines synthetase is investigated. Results indicate that the knockout mutant is impaired in nitrogen assimilation and translocation under salt treatment
malfunction
-
silencing of CaAS1 gene results in enhanced susceptibility to Xanthomonas campestris pv. vesicatoria infection, phenotypes, overview
malfunction
Q2L9S5
silencing of the BmASNS gene enhances the sensitivity of silkworm cells to amino acid starvation. Ectopic overexpression of BmASNS gene effectively inhibits cell growth in silkworm cells, whereas its overexpression can rescue cell growth upon amino acid deprivation treatment. Silkworm cells lacking BmASNS under the condition of amino acid deprivation show severely impaired proliferation
physiological function
-
Arabidopsis knockout mutant with T-DNA insertion in ASN2 gene, subject to 100 mM NaCl stress for 6 to 24 h. The salt treatment decreases chlorophyll and soluble protein contents, and increases ammonium level in the asn2-1 leaves. The salinity induces ASN1 mRNA level in the wild-type and asn2-1 leaves. The salt treatment inhibits the transcript and protein levels of chloroplastic glutamine synthetase 2, EC 6.3.1.2 in the wild-type and asn2-1 leaves
physiological function
-
silencing of the gene encoding AS1 results in enhanced susceptibility to Xanthomonas campestris pv. vesicatoria infection. Transgenic Arabidopsis thaliana plants that overexpress CaAS1 exhibit enhanced resistance to Pseudomonas syringae pv. tomato DC3000 and Hyaloperonospora arabidopsidis. Increased CaAS1 expression influences early defense responses in diseased leaves, including increased electrolyte leakage, reactive oxygen species and nitric oxide burst. In CaAS1-silenced pepper and/or CaAS1-overexpressing Arabidopsis, CaAS1-dependent changes in asparagine levels correlate with increased susceptibility or defense responses to microbial pathogens, respectively, asparagine synthetase 1 is essential for plant defense to microbial pathogens. Increased CaAS1 expression influences early defense responses in diseased leaves, including increased electrolyte leakage, reactive oxygen species and nitric oxide bursts. In plants, increased conversion of aspartate to asparagine appears to be associated with enhanced resistance to bacterial and oomycete pathogens, phenotypes, overview
physiological function
Q2L9S5
BmAsns protein negatively regulates silkworm cell proliferation. The recovery of cell growth by overexpressed BmAsns protein is due to the rapid turnover of autophagic vacuoles in the cells
physiological function
-
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
physiological function
D9IXC8, E7EAQ2
the expression of PpAS1 is regulated by developmental and environmental factors
metabolism
-
Asn is a major amino acid in maize and since AsnS is the primary means of Asn synthesis in plants it plays a very important role in nitrogen metabolism
additional information
-
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
additional information
-
transgenic Arabidopsis thaliana plants that overexpress CaAS1 exhibit enhanced resistance to Pseudomonas syringae pv. tomato DC3000 and Hyaloperonospora arabidopsidis
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
ATP + cysteine sulfinic acid
AMP + diphosphate + cysteine sulfinic acid
show the reaction diagram
-
-
-
?
ATP + L-Asp + L-Gln
AMP + diphosphate + Asn + Glu
show the reaction diagram
-
-
-
-
-
ATP + L-Asp + L-Gln
AMP + diphosphate + Asn + Glu
show the reaction diagram
-
-
-
-
-
ATP + L-Asp + L-Gln
AMP + diphosphate + Asn + Glu
show the reaction diagram
-
-
-
-
-
ATP + L-Asp + L-Gln
AMP + diphosphate + Asn + Glu
show the reaction diagram
-
-
-
-
-
ATP + L-Asp + L-Gln
AMP + diphosphate + Asn + Glu
show the reaction diagram
-
-
-
-
-
ATP + L-Asp + L-Gln
AMP + diphosphate + Asn + Glu
show the reaction diagram
-
-
-
-
-
ATP + L-Asp + L-Gln
AMP + diphosphate + Asn + Glu
show the reaction diagram
-
-
-
-
?
ATP + L-Asp + L-Gln
AMP + diphosphate + Asn + Glu
show the reaction diagram
-
-
-
-
-
ATP + L-Asp + L-Gln
AMP + diphosphate + Asn + Glu
show the reaction diagram
-
-
-
-
ATP + L-Asp + L-Gln
AMP + diphosphate + Asn + Glu
show the reaction diagram
-
-
-
-
-
ATP + L-Asp + L-Gln
AMP + diphosphate + Asn + Glu
show the reaction diagram
-
-
-
-
-
ATP + L-Asp + L-Gln
AMP + diphosphate + Asn + Glu
show the reaction diagram
-
-
-
-
-
ATP + L-Asp + L-Gln
AMP + diphosphate + Asn + Glu
show the reaction diagram
-
-
-
-
-
ATP + L-Asp + L-Gln
AMP + diphosphate + Asn + Glu
show the reaction diagram
-
-
-
-
-
ATP + L-Asp + L-Gln
AMP + diphosphate + Asn + Glu
show the reaction diagram
-
-
-
-
-
ATP + L-Asp + L-Gln
AMP + diphosphate + Asn + Glu
show the reaction diagram
-
-
-
-
-
ATP + L-Asp + L-Gln
AMP + diphosphate + Asn + Glu
show the reaction diagram
-
-
-
-
-
ATP + L-Asp + L-Gln
AMP + diphosphate + Asn + Glu
show the reaction diagram
-
-
-
-
-
ATP + L-Asp + L-Gln
AMP + diphosphate + Asn + Glu
show the reaction diagram
-
-
-
-
-
ATP + L-Asp + L-Gln
AMP + diphosphate + Asn + Glu
show the reaction diagram
-
-
-
-
-
ATP + L-Asp + L-Gln
AMP + diphosphate + Asn + Glu
show the reaction diagram
-
-
-
-
-
ATP + L-Asp + L-Gln
AMP + diphosphate + Asn + Glu
show the reaction diagram
-
-
-
-
-
ATP + L-Asp + L-Gln
AMP + diphosphate + Asn + Glu
show the reaction diagram
-
-
-
-
-
ATP + L-Asp + L-Gln
AMP + diphosphate + Asn + Glu
show the reaction diagram
-
-
-
-
-
ATP + L-Asp + L-Gln
AMP + diphosphate + Asn + Glu
show the reaction diagram
-
-
-
-
-
ATP + L-Asp + L-Gln
AMP + diphosphate + Asn + Glu
show the reaction diagram
-
-
-
-
-
ATP + L-Asp + L-Gln
AMP + diphosphate + Asn + Glu
show the reaction diagram
Q8KN11
-
-
-
?
ATP + L-Asp + L-Gln
AMP + diphosphate + Asn + Glu
show the reaction diagram
-
ir
-
-
-
ATP + L-Asp + L-Gln
AMP + diphosphate + Asn + Glu
show the reaction diagram
-
the ratio of Gln- to NH4+-dependent activity is 2.5
-
-
-
ATP + L-Asp + L-Gln
AMP + diphosphate + Asn + Glu
show the reaction diagram
-
TaASN1 is dramatically induced by salinity, osmotic stress and exogenous abscisic acid, TaASN2 transcripts are very low in all detected tissues and conditions and are only slightly induced by abscisic acid in roots
-
-
?
ATP + L-Asp + L-Gln
AMP + diphosphate + Asn + Glu
show the reaction diagram
-
the basic region leucine zipper protein ATF5, a transcriptional activator, stimulates asparagine promoter/reporter gene transcription via the nutrient-sensing response unit
-
-
?
ATP + L-Asp + L-Gln
AMP + diphosphate + Asn + Glu
show the reaction diagram
-
resistance to L-asparaginase and relapse risk are associated with high expression of asparagine synthetase in TEL-AML1-negative but not in TEL-AML1-positive B-lineage acute lymphoblastic leukemia
-
-
?
ATP + L-Asp + L-Gln
AMP + diphosphate + L-Asn + L-Glu
show the reaction diagram
-
-
-
-
?
ATP + L-Asp + L-Gln
AMP + diphosphate + L-Asn + L-Glu
show the reaction diagram
-
-
-
-
?
ATP + L-Asp + L-Gln
AMP + diphosphate + L-Asn + L-Glu
show the reaction diagram
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
-
-
-
?
ATP + L-Asp + L-Gln
AMP + diphosphate + L-Asn + L-Glu
show the reaction diagram
-
light, carbon and nitrogen availability control asparagine synthesis in sunflower by regulating three aspargine synthetase coding genes. HAS2 expression requires light and is positively affected by sucrose. HAS1 and HAS1.1 expression is dependent on nitrogen availability, while HAS2 transcripts are still found in N-starved plants. High ammonium level induces all three asparagine synthetase genes and partially reverts sucrose repression of HAS1 and HAS1.1
-
-
?
ATP + L-Asp + L-Gln + H2O
AMP + diphosphate + L-Asn + L-Glu
show the reaction diagram
-
-
-
-
?
ATP + L-Asp + NH2OH
AMP + diphosphate + beta-aspartylhydroxamate
show the reaction diagram
-
-
-
-
ATP + L-Asp + NH2OH
AMP + diphosphate + beta-aspartylhydroxamate
show the reaction diagram
-
-
-
-
-
ATP + L-Asp + NH2OH
AMP + diphosphate + beta-aspartylhydroxamate
show the reaction diagram
-
-
-
-
ATP + L-Asp + NH2OH
AMP + diphosphate + beta-aspartylhydroxamate
show the reaction diagram
-
-
-
-
ATP + L-Asp + NH2OH
AMP + diphosphate + beta-aspartylhydroxamate
show the reaction diagram
-
-
-
-
-
ATP + L-Asp + NH2OH
AMP + diphosphate + beta-aspartylhydroxamate
show the reaction diagram
-
-
-
-
-
ATP + L-Asp + NH3
AMP + diphosphate + Asn
show the reaction diagram
-
-
-
-
-
ATP + L-Asp + NH3
AMP + diphosphate + Asn
show the reaction diagram
-
-
-
-
-
ATP + L-Asp + NH3
AMP + diphosphate + Asn
show the reaction diagram
-
-
-
-
?
ATP + L-Asp + NH3
AMP + diphosphate + Asn
show the reaction diagram
-
-
-
-
-
ATP + L-Asp + NH3
AMP + diphosphate + Asn
show the reaction diagram
-
-
-
-
-
ATP + L-Asp + NH3
AMP + diphosphate + Asn
show the reaction diagram
-
-
-
-
-
ATP + L-Asp + NH3
AMP + diphosphate + Asn
show the reaction diagram
-
-
-
-
-
ATP + L-Asp + NH3
AMP + diphosphate + Asn
show the reaction diagram
-
the ratio of Gln-dependent to NH4+-dependent activity is 2.5
-
-
-
ATP + L-Asp + NH3
AMP + diphosphate + Asn
show the reaction diagram
-
85% of the activity relative to Gln
-
-
-
ATP + L-Asp + NH3
AMP + diphosphate + Asn
show the reaction diagram
-
30% of the activity relative to Gln
-
-
-
ATP + L-Asp + NH3
AMP + diphosphate + Asn
show the reaction diagram
-
NH4+
-
-
-
ATP + L-Asp + NH3
AMP + diphosphate + Asn
show the reaction diagram
-
NH4+
-
-
-
ATP + L-Asp + NH3
AMP + diphosphate + L-Asn
show the reaction diagram
-
-
-
-
?
ATP + L-Asp + NH3
AMP + diphosphate + L-Asn
show the reaction diagram
-
-
-
-
?
ATP + L-Asp + NH3
AMP + diphosphate + L-Asn
show the reaction diagram
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
-
-
-
?
ATP + L-aspartate + L-glutamine
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
-
-
-
?
ATP + L-aspartate + L-glutamine
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
-
-
-
-
?
ATP + L-aspartate + L-glutamine
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
-
-
-
?
ATP + L-aspartate + L-glutamine
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
-
-
-
?
ATP + L-aspartate + L-glutamine
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
-
-
-
?
ATP + L-aspartate + L-glutamine
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
P22106
-
-
?
ATP + L-aspartate + L-glutamine
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
-
-
-
?
ATP + L-aspartate + L-glutamine
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
-
-
-
?
ATP + L-aspartate + L-glutamine
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
-
-
-
?
ATP + L-aspartate + L-glutamine
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
-
-
-
?
ATP + L-aspartate + L-glutamine
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
-
-
-
?
ATP + L-aspartate + L-glutamine
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
-
-
-
?
ATP + L-aspartate + L-glutamine
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
Q84LA5, Q93XP9
-
-
?
ATP + L-aspartate + L-glutamine
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
Q9SM55
-
-
?
ATP + L-aspartate + L-glutamine
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
-
ATP-dependent, mechanism including an enzyme-ATP-Asp-Gln quarternary complex, AS-B structure, two active sites
-
ir
ATP + L-aspartate + L-glutamine
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
-
ATP-dependent, the amine group of Gln is transferred directly to Asp
-
?
ATP + L-aspartate + L-glutamine
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
-
ATP-dependent, the amine group of Gln is transferred directly to Asp, maximum activity with 1 mM Gln and 3-10 mM ATP in the assay
-
?
ATP + L-aspartate + L-glutamine
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
-
transfers the amide group of Gln to Asp
-
?
ATP + L-aspartate + L-glutamine
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
-
glutamine is the in vivo nitrogen source
-
?
ATP + L-aspartate + L-glutamine
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
-
the enzyme might play a functional role in nitrogen translocation from root to aerial organs in Phaseolus vulgaris
-
-
?
ATP + L-aspartate + L-glutamine
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
-
the reaction sequence begins with the ordered addition of ATP and aspartate. Diphosphate is released, followed by the addition of ammonia and the release of asparagine and AMP. Glutamine is simultaneously hydrolyzed at a second site and the ammonia intermediate diffuses through an interdomain protein tunnel from the site of production to the site of utilization. The data are also consistent with the dead-end binding of asparagine to the glutamine binding site and diphosphate with free enzyme. The rate of hydrolysis of glutamine is largely independent of the activation of aspartate and thus the reaction rates at the two active sites are essentially uncoupled from one another
-
-
?
ATP + L-aspartate + L-glutamine
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
Bacillus subtilis 168
-
-
-
?
ATP + L-aspartate + L-glutamine + H2O
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
-
-
-
-
?
ATP + L-aspartate + L-glutamine + H2O
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
A9XS73
-
-
-
?
ATP + L-aspartate + L-glutamine + H2O
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
-
-
-
?
ATP + L-aspartate + L-glutamine + H2O
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
Q2L9S5
-
-
-
?
ATP + L-aspartate + L-glutamine + H2O
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
D9IXC8, E7EAQ2
-
-
-
?
ATP + L-aspartate + NH3
AMP + diphosphate + L-asparagine
show the reaction diagram
P22106
-
-
?
ATP + L-aspartate + NH3
AMP + diphosphate + L-asparagine
show the reaction diagram
-
asparagine biosynthesis proceeds by initial reaction of aspartate and ATP to yield a beta-aspartyl-AMP intermediate, in the presence of glutamine, ammonia released in the N-terminal active site reacts with beta-aspartyl-AMP to yield asparagine and AMP
-
?
ATP + L-aspartate + NH3
AMP + diphosphate + L-asparagine
show the reaction diagram
-
asparagine biosynthesis proceeds by initial reaction of aspartate and ATP to yield a beta-aspartyl-AMP intermediate, in the presence of glutamine, ammonia released in the N-terminal active site reacts with beta-aspartyl-AMP to yield asparagine and AMP
-
?
ATP + L-aspartate + NH3
AMP + diphosphate + L-asparagine
show the reaction diagram
-
asparagine biosynthesis proceeds by initial reaction of aspartate and ATP to yield a beta-aspartyl-AMP intermediate, in the presence of glutamine, ammonia released in the N-terminal active site reacts with beta-aspartyl-AMP to yield asparagine and AMP
-
?
ATP + L-aspartate + NH3
AMP + diphosphate + L-asparagine
show the reaction diagram
-
asparagine biosynthesis proceeds by initial reaction of aspartate and ATP to yield a beta-aspartyl-AMP intermediate, in the presence of glutamine, ammonia released in the N-terminal active site reacts with beta-aspartyl-AMP to yield asparagine and AMP
-
?
CTP + L-Asp + L-Gln
CMP + diphosphate + Asn + Glu
show the reaction diagram
-
weak activity
-
-
-
dATP + L-Asp + L-Gln
dAMP + diphosphate + Asn + Glu
show the reaction diagram
-
utilized at a similar rate as ATP
-
-
-
dATP + L-Asp + NH3
dAMP + diphosphate + Asn
show the reaction diagram
-
utilized at a similar rate as ATP
-
-
-
GTP + L-Asp + L-Gln
GMP + diphosphate + Asn + Glu
show the reaction diagram
-
5.6% of the activity relative to ATP
-
-
-
GTP + L-Asp + L-Gln
GMP + diphosphate + Asn + Glu
show the reaction diagram
-
15% of the activity relative to ATP
-
-
-
L-Glutamic acid gamma-monohydroxamate + H2O
Hydroxylamine + Glu
show the reaction diagram
-
-
-
-
-
L-glutamine
L-glutamate + NH3
show the reaction diagram
-
-
-
?
L-glutamine
L-glutamate + NH3
show the reaction diagram
-
-
-
?
L-glutamine
L-glutamate + NH3
show the reaction diagram
P22106
-
-
?
L-glutamine
L-glutamate + NH3
show the reaction diagram
-
-
-
?
UTP + L-Asp + L-Gln
UMP + diphosphate + Asn + Glu
show the reaction diagram
-
weak activity
-
-
-
L-glutamine
L-glutamate + NH3
show the reaction diagram
-
-
-
?
additional information
?
-
-
the N74D As-B mutant exhibits very low glutaminase activity
-
-
-
additional information
?
-
-
no activity with ITP or XTP
-
-
-
additional information
?
-
-
enzyme is extremly selective, being able to discriminate between metabolites that have similar structures to Asp
-
-
-
additional information
?
-
-
ATP-diphosphate exchange reaction
-
-
-
additional information
?
-
-
ATP-diphosphate exchange reaction
-
-
-
additional information
?
-
-
the synthesis of Asn in mammalian tissues proceeds through the intermediate beta-aspartyladenylate
-
-
-
additional information
?
-
-
enzyme has inherent glutaminase activity
-
-
-
additional information
?
-
-
enzyme has inherent glutaminase activity
-
-
-
additional information
?
-
-
enzyme has inherent glutaminase activity
-
-
-
additional information
?
-
-
enzyme has inherent glutaminase activity
-
-
-
additional information
?
-
-
possible involvement of the enzyme in the control of metabolic fluxes of carbon and nitrogen through assimilatory pathways
-
-
-
additional information
?
-
-
enzyme is involved in ammonia assimilation
-
-
-
additional information
?
-
-
Asn, the end product of the mass action of symbiotic NH4+ synthesis, is the principal N-transport-compound of many temperate legumes
-
-
-
additional information
?
-
-
the primary site of Asn synthesis is the root and subsequently the leaves receive Asn as the principal N-source for amino acid and protein synthesis
-
-
-
additional information
?
-
-
importance of asparagine synthetase in cell proliferation
-
-
-
additional information
?
-
-
Gln-dependent enzyme is essential for Asn synthesis when the nitrogen source is growth rate limiting
-
-
-
additional information
?
-
-
comprehensive mechanism has been proposed through which either Gln or NH3 can provide nitrogen for Asn production from Asp
-
-
-
additional information
?
-
-
glutamine or glutamine-derived metabolites regulate AS expression in roots
-
?
additional information
?
-
-
major biosynthetic pathway for asparagine, AS gene expression is down-regulated by light
-
?
additional information
?
-
-
nitrogen metabolism, asparagine synthesis
-
?
additional information
?
-
-
physiological roles for asnB in vegetative cells and for asnO in sporulating cells, asnB may be the main gene involved in asparagine biosynthesis
-
?
additional information
?
-
-
primary enzyme responsible for asparagine synthesis
-
?
additional information
?
-
Q84LA5, Q93XP9
under normal growth conditions HvAS1 gene seems to be important in roots where nitrogen is assimilated into asparagine for long-distance transport within the plant
-
?
additional information
?
-
Q84LA5, Q93XP9
under normal growth conditions HvAS2 acts as a housekeeping gene in the leaves
-
?
additional information
?
-
Q9LKQ9, Q9SP19, Q9SP20
Helianthus annuus contains three asparagine synthetase genes: HAS1, HAS1.1 and HAS2. Most of the asparagine newly synthesized for germination and cotyledon expansion is due to HAS2 activity, with little contribution of the other asparagine synthetase genes. All three genes work together to synthesize asparagine for leaf senescence
-
-
-
additional information
?
-
-
upregulation of asparagine synthetase fails to avert cell cycle arrest induced by L-asparaginase in TEL/AML1-positive leukaemic cells
-
-
-
additional information
?
-
A9XS73
role for hexokinase in the sugar-sensing mechanism that regulates PvNAS2 expression in roots, downregulation of the asparagine synthetase enzyme and concomitantly asparagine production. Thereby a favourable environment is created for the efficient transfer of the amido group of glutamine for the synthesis of purines, and then ureide generation.
-
-
-
additional information
?
-
Bacillus subtilis 168
-
physiological roles for asnB in vegetative cells and for asnO in sporulating cells, asnB may be the main gene involved in asparagine biosynthesis
-
?
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
ATP + L-Asp + L-Gln
AMP + diphosphate + Asn + Glu
show the reaction diagram
-
TaASN1 is dramatically induced by salinity, osmotic stress and exogenous abscisic acid, TaASN2 transcripts are very low in all detected tissues and conditions and are only slightly induced by abscisic acid in roots
-
-
?
ATP + L-Asp + L-Gln
AMP + diphosphate + Asn + Glu
show the reaction diagram
-
the basic region leucine zipper protein ATF5, a transcriptional activator, stimulates asparagine promoter/reporter gene transcription via the nutrient-sensing response unit
-
-
?
ATP + L-Asp + L-Gln
AMP + diphosphate + L-Asn + L-Glu
show the reaction diagram
-
light, carbon and nitrogen availability control asparagine synthesis in sunflower by regulating three aspargine synthetase coding genes. HAS2 expression requires light and is positively affected by sucrose. HAS1 and HAS1.1 expression is dependent on nitrogen availability, while HAS2 transcripts are still found in N-starved plants. High ammonium level induces all three asparagine synthetase genes and partially reverts sucrose repression of HAS1 and HAS1.1
-
-
?
ATP + L-aspartate + L-glutamine
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
-
-
-
?
ATP + L-aspartate + L-glutamine
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
-
-
-
?
ATP + L-aspartate + L-glutamine
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
-
-
-
?
ATP + L-aspartate + L-glutamine
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
P22106
-
-
?
ATP + L-aspartate + L-glutamine
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
-
-
-
?
ATP + L-aspartate + L-glutamine
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
-
-
-
?
ATP + L-aspartate + L-glutamine
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
-
-
-
?
ATP + L-aspartate + L-glutamine
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
-
-
-
?
ATP + L-aspartate + L-glutamine
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
-
-
-
?
ATP + L-aspartate + L-glutamine
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
-
glutamine is the in vivo nitrogen source
-
?
ATP + L-aspartate + L-glutamine
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
-
the enzyme might play a functional role in nitrogen translocation from root to aerial organs in Phaseolus vulgaris
-
-
?
ATP + L-aspartate + L-glutamine + H2O
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
-
-
-
-
?
ATP + L-aspartate + L-glutamine + H2O
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
A9XS73
-
-
-
?
ATP + L-aspartate + L-glutamine + H2O
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
-
-
-
?
ATP + L-aspartate + L-glutamine + H2O
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
Q2L9S5
-
-
-
?
ATP + L-aspartate + L-glutamine + H2O
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
D9IXC8, E7EAQ2
-
-
-
?
ATP + L-aspartate + NH3
AMP + diphosphate + L-asparagine
show the reaction diagram
P22106
-
-
?
ATP + L-aspartate + NH3
AMP + diphosphate + L-asparagine
show the reaction diagram
-
asparagine biosynthesis proceeds by initial reaction of aspartate and ATP to yield a beta-aspartyl-AMP intermediate, in the presence of glutamine, ammonia released in the N-terminal active site reacts with beta-aspartyl-AMP to yield asparagine and AMP
-
?
ATP + L-aspartate + NH3
AMP + diphosphate + L-asparagine
show the reaction diagram
-
asparagine biosynthesis proceeds by initial reaction of aspartate and ATP to yield a beta-aspartyl-AMP intermediate, in the presence of glutamine, ammonia released in the N-terminal active site reacts with beta-aspartyl-AMP to yield asparagine and AMP
-
?
ATP + L-aspartate + NH3
AMP + diphosphate + L-asparagine
show the reaction diagram
-
asparagine biosynthesis proceeds by initial reaction of aspartate and ATP to yield a beta-aspartyl-AMP intermediate, in the presence of glutamine, ammonia released in the N-terminal active site reacts with beta-aspartyl-AMP to yield asparagine and AMP
-
?
ATP + L-aspartate + NH3
AMP + diphosphate + L-asparagine
show the reaction diagram
-
asparagine biosynthesis proceeds by initial reaction of aspartate and ATP to yield a beta-aspartyl-AMP intermediate, in the presence of glutamine, ammonia released in the N-terminal active site reacts with beta-aspartyl-AMP to yield asparagine and AMP
-
?
L-glutamine
L-glutamate + NH3
show the reaction diagram
-
-
-
?
L-glutamine
L-glutamate + NH3
show the reaction diagram
-
-
-
?
L-glutamine
L-glutamate + NH3
show the reaction diagram
P22106
-
-
?
L-glutamine
L-glutamate + NH3
show the reaction diagram
-
-
-
?
L-glutamine
L-glutamate + NH3
show the reaction diagram
-
-
-
?
additional information
?
-
-
possible involvement of the enzyme in the control of metabolic fluxes of carbon and nitrogen through assimilatory pathways
-
-
-
additional information
?
-
-
enzyme is involved in ammonia assimilation
-
-
-
additional information
?
-
-
Asn, the end product of the mass action of symbiotic NH4+ synthesis, is the principal N-transport-compound of many temperate legumes
-
-
-
additional information
?
-
-
the primary site of Asn synthesis is the root and subsequently the leaves receive Asn as the principal N-source for amino acid and protein synthesis
-
-
-
additional information
?
-
-
importance of asparagine synthetase in cell proliferation
-
-
-
additional information
?
-
-
Gln-dependent enzyme is essential for Asn synthesis when the nitrogen source is growth rate limiting
-
-
-
additional information
?
-
-
comprehensive mechanism has been proposed through which either Gln or NH3 can provide nitrogen for Asn production from Asp
-
-
-
additional information
?
-
-
glutamine or glutamine-derived metabolites regulate AS expression in roots
-
?
additional information
?
-
-
major biosynthetic pathway for asparagine, AS gene expression is down-regulated by light
-
?
additional information
?
-
-
nitrogen metabolism, asparagine synthesis
-
?
additional information
?
-
-
physiological roles for asnB in vegetative cells and for asnO in sporulating cells, asnB may be the main gene involved in asparagine biosynthesis
-
?
additional information
?
-
-
primary enzyme responsible for asparagine synthesis
-
?
additional information
?
-
Q84LA5, Q93XP9
under normal growth conditions HvAS1 gene seems to be important in roots where nitrogen is assimilated into asparagine for long-distance transport within the plant
-
?
additional information
?
-
Q84LA5, Q93XP9
under normal growth conditions HvAS2 acts as a housekeeping gene in the leaves
-
?
additional information
?
-
Q9LKQ9, Q9SP19, Q9SP20
Helianthus annuus contains three asparagine synthetase genes: HAS1, HAS1.1 and HAS2. Most of the asparagine newly synthesized for germination and cotyledon expansion is due to HAS2 activity, with little contribution of the other asparagine synthetase genes. All three genes work together to synthesize asparagine for leaf senescence
-
-
-
additional information
?
-
-
upregulation of asparagine synthetase fails to avert cell cycle arrest induced by L-asparaginase in TEL/AML1-positive leukaemic cells
-
-
-
additional information
?
-
A9XS73
role for hexokinase in the sugar-sensing mechanism that regulates PvNAS2 expression in roots, downregulation of the asparagine synthetase enzyme and concomitantly asparagine production. Thereby a favourable environment is created for the efficient transfer of the amido group of glutamine for the synthesis of purines, and then ureide generation.
-
-
-
additional information
?
-
Bacillus subtilis 168
-
physiological roles for asnB in vegetative cells and for asnO in sporulating cells, asnB may be the main gene involved in asparagine biosynthesis
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
ATP
A9XS73
-
ATP
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
-
ATP
Q2L9S5
-
ATP
D9IXC8, E7EAQ2
;
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Br-
-
strong activation of Asn synthesis and Gln hydrolysis. The synthetase reactions with NH3 or NH2OH are only slightly stimulated
Br-
-
can partially replace Cl- in activation
Ca2+
-
at 16.7 mM CaCl2 31% of the activation relative to MgCl2
Cl-
-
stimulation of the Gln-dependent reaction and glutaminase reaction. Inhibition of the NH4+-dependent reaction, competitive with respect to NH4+, with negative cooperativity
Cl-
-
strong activation of Asn synthesis and Gln hydrolysis, competitive activator with respect to Gln and a noncompetitive activator with respect to MgATP2- and Asp. Cl- changes the substrate saturation kinetics of Gln from negatively cooperative to normal hyperbolic and causes a 50fold increase in the affinity for Gln. Inherent glutaminase activity is enhanced 30fold. The synthetase reactions with NH4+ or NH2OH are only slightly stimulated
Cl-
-
required, Km: 1.7 mM
Cl-
-
required for glutaminase and glutamine-dependent synthetase activity, not for the NH4+-dependent activity
CN-
-
stimulates Asn synthesis and Gln hydrolysis
Co2+
-
at 16.7 mM CoCl2 31% of the activation relative to MgCl2
Co2+
-
is the only divalent metal ion that can replace Mg2+ with 80% of the Mg2+-dependent AS-B activity activity with Gln and 52% of the Mg2+-dependent AS-B activity with NH4+
Co2+
-
can partially replace Mg2+ in activation
Cu2+
-
0.02 mM, induces expression of the enzyme
F-
-
stimulates Asn synthesis and Gln hydrolysis
FeCl2
-
at 16.7 mM FeCl2 39% of the activation relative to MgCl2
I-
-
stimulates Asn synthesis and Gln hydrolysis
I-
-
can partially replace Cl- in activation
Mg2+
-
forms a complex with ATP and modifies the reaction mechanism
Mg2+
-
16.7 mM MgCl2, activates
Mg2+
-
highest stimulation of divalent metal ions
Mg2+
-
Km: 1.1 mM
Mg2+
-
divalent metal ion required, Mg2+ more effective than Mn2+
Mg2+
-
Mg2+ cannot be replaced by Zn2+, Fe2+, Co2+, Ba2+ or Ca2+
Mg2+
-
-
Mg2+
A9XS73
-
Mg2+
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
required; required; required; required
Mg2+
-
divalent metal ion required, Mg2+ more effective than Mn2+; Km: 23.7
Mn2+
-
at 16.7 mM MnCl2 14% of the activation relative to MgCl2
Mn2+
-
can partially substitute for Mg2+
Mn2+
-
10% of the activity relative to Mg2+
Mn2+
-
3 mM or 10 mM Mn2+ gives 9% and 6% of the activity relative to Mg2+
Mn2+
-
at 2 mM 22.0% of the activity relative to Mg2+
Na+
-
150 mM, induces expression of the enzyme
Ni2+
-
at 16.7 mM NiCl2 17% of the activation relative to MgCl2
Ni2+
-
can partially replace Mg2+ in activation
NO3-
-
stimulates Asn synthesis and Gln hydrolysis
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
1-methyl-4-(1-methylethyl)-7-oxabicyclo[2.2.1]heptane
-
trivial name 1,4-cineole, almost complete inhibition above 1 mM
2,3-dicarboxypyridine
-
-
2,4-Dicarboxypyridine
-
-
2,5-Dicarboxypyridine
-
-
2,6-dicarboxypyridine
-
-
2-Amino-2-carboxy-L-ethanesulfonamide
-
-
2-Amino-4-arsonophenol hydrochloride
-
-
2-carboxypyridine
-
-
2-Hydroxyethyl-L-Gln
-
-
2-oxoglutarate
-
20 mM, 12-15% inhibition
2-oxoglutarate
-
in presence of aminoxyacetate, 50% inhibition at 5 mM
3,4-Dicarboxypyridine
-
-
3,5-Dicarboxypyridine
-
-
4-Carboxypyridine
-
-
5'-O-[p-(fluorosulfonyl)benzoyl]adenosine
-
covalently modifies the enzyme
5'-O-[p-(fluorosulfonyl)benzoyl]adenosine
-
Cys523 is the key residue involved in the formation of the 5'-O-[p-(fluorosulfonyl)benzoyl]adenosine-induced disulfide bond, inactivation can be reversed by addition of dithiothreitol
5-Bromo-4-oxo-L-norvaline
-
-
5-Chloro-4-oxo-L-norvaline
-
-
5-Chloro-4-oxo-L-norvaline
-
-
5-Chloro-4-oxo-L-norvaline
-
-
5-Diazo-4-oxo-L-norvaline
-
-
6-diazo-5-oxo-L-norleucine
-
-
6-diazo-5-oxo-L-norleucine
-
loss of Gln-dependent reactions, but no effect on ATP binding as measured during amminoa-dependent Asn synthesis
6-diazo-5-oxo-L-norleucine
-
-
8-N3ATP
-
loss of NH4+-dependent Asn synthesis, but no effect on the glutaminase activity
Adenosine-5'-methylphosphonate
-
-
adenylated sulfoximine
-
0.005 mM, 65% inhibition, dead-end complex with AS-B
ADP
-
weak
Albizzine
-
-
Albizzine
-
-
Albizzine
-
-
alpha,beta-methylene ATP
-
-
alpha,beta-methylene-ATP
-
-
Aminomalonic acid
-
-
Aminomalonic acid
-
competitive versus L-Asp
ammonium maleamate
-
weak
AMP
-
product inhibition
AMP
-
noncompetitive versus ATP; poor inhibitor
AMP
-
weak
AMP
-
2.5 mM, 50% inhibition
AMP-PNP
-
competitive vs. ATP, noncompetitive vs. aspartate, uncompetitive vs. glutamine
Arg
-
-
Asn
-
product inhibition
Asn
-
-
asparagine
-
noncompetitive vs. ATP and aspartate
asparagine
-
-
asparagine
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
competitive; competitive
aspartic acid analogs
-
-
ATP
-
strong inhibition above 5 mM
azaserine
-
-
azaserine
-
-
azaserine
-
-
azaserine
-
-
beta,gamma-methylene ATP
-
-
beta-asparaginyladenylate
-
-
beta-Methyl-DL-Asn
-
-
beta-methylaspartate
-
weak inhibitor
beta-methylaspartate
-
noncompetitive vs. ATP, competitive vs. aspartate, noncompetitive vs. glutamine
Ca2+
-
-
Ca2+
-
-
Carbamoyl phosphate
-
-
cis-2-hydroxy-1,4-cineole
-
0.00003 mM, 50% inhibition
Cl-
-
inhibition of the ammonia-dependent reaction, competitive with respect to ammonia, with negative cooperativity. Stimulation of the Gln-dependent and glutaminase reaction
CMP
-
weak
Cu2+
-
-
D-Aspartic acid
-
weak
diphosphate
-
competitive versus ATP
diphosphate
-
5 mM, 45% inhibition
diphosphate
-
noncompetitive vs. ATP
diphosphate
-
-
diphosphate
-
-
diphosphate
-
-
diphosphate
-
-
DL-alpha-Aminotricarballylic acid
-
weak
DL-homo-Gln
-
-
erythro-beta-Hydroxy-L-Asn
-
-
erythro-beta-hydroxy-L-Asp
-
-
erythro-beta-Methyl-L-Asp
-
-
gamma-Methylene-L-Gln
-
-
GDP
-
weak
Gln
-
0.4-2.0 mM, inhibits the ammonia-dependent production of Asn
Gln
-
inhibitor of the NH4+-dependent reaction catalyzed by both the C1A and C1S mutants of AS-B
Gln
-
D-Gln, weak
Glu
-
product inhibition
Glu
-
poor inhibitor; product inhibition
Glu
-
150 mM, 50% inhibition
Glu
-
-
glutamate
-
-
glutamate
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
competitive; competitive
Gly-L-Asn
-
weak
GMP
-
weak
IMP
-
weak
iodoacetamide
-
-
L-(alphaS,5S)-alpha-Amino-3-chloro-4,5-dihydroisoxazol-5-ylacetic acid
-
i.e. NSC-163501
L-1-Amino-4-oxo-5-(5'-adenosyl)phosphopentanoic acid
-
noncompetitive with respect to Gln and uncompetitive with respect to both ATP and Asp
-
L-2-Amino-4-oxo-5-chloropentanoic acid
-
-
L-2-Amino-4-oxo-5-hydroxypentanoic acid
-
-
L-2-Amino-4-oxo-5-methylphosphonopentanoic acid
-
-
L-asparagine
-
product inhibition
L-asparagine
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
competitive inhibition
L-beta-Aspartate ethyl ester
-
-
L-cysteinesulfinic acid
-
-
L-glutamate
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
competitive inhibition
L-Glutamate diamide
-
-
L-Glutamate-gamma-ethyl ester
-
-
L-glutamate-gamma-methyl ester
-
-
L-glutamic acid gamma-methyl ester
-
-
L-glutamic acid gamma-methylester
-
uncompetitive vs. ATP, competitive vs. glutamine
L-Homoserine beta-adenylate
-
in the presence of 30 mM MgCl2
L-Malic acid
-
weak
L-methionine sulfoxide
-
-
L-methionine-S-sulfoximine
-
-
Maleimide
-
-
meso-Diaminosuccinamate
-
-
Mucochloric acid
-
-
mupirocin
-
-
N-acetyl-L-Gln
-
-
N-alpha-Methyl-L-Asn
-
-
N-Benzyloxycarbonyl-L-Asn
-
weak
N-Benzyloxycarbonyl-L-aspartate
-
weak
N-Carbobenzoxy-DL-Gln
-
-
N-Ethyl-L-Asn
-
-
N-Methyl-DL-aspartic acid
-
-
N-Methyl-L-Asn
-
-
nucleoside triphosphates
-
except ATP
O-acetylserine
-
-
oxaloacetate
-
20 mM, 40% inhibition
p-chloromercuribenzoate
-
-
phosmidosine
-
-
Phthalic acid
-
weak
pyrrolidine-2,3-dicarboxylic acid
-
weak inhibitor
pyruvate
-
20 mM, 12-15% inhibition
S-Carbamoylcysteine
-
-
S-Carbamoylcysteine
-
-
S-Carbamoylcysteine
-
-
S-methyl-L-cysteine
-
-
S-methyl-L-cysteine-(RS)-sulfoximine
-
weak
Sn2+
-
-
Sr2+
-
-
sulfhydryl reagents
-
-
sulfoximine adenylate
-
most potent inhibitor
threo-beta-Hydroxy-L-Asn
-
-
threo-beta-methyl-L-Asp
-
-
trans-2-hydroxy-1,4-cineole
-
0.01 mM, 50% inhibition
Trp
-
-
UMP
-
weak
Zn2+
-
ZnCl2
Zn2+
-
-
Methyl gamma-Gln
-
-
additional information
-
mouse pancreas contains a proteolytic inhibitor of L-Asn synthetase
-
additional information
-
enzyme from fetal liver extracts is significantly inhibited when combined with adult liver or tumor extracts
-
additional information
-
methionine sulfoximine completely inhibits the NH4+-induced accumulation of AS protein, but not the glutamine-induced accumulation
-
additional information
-
L-asparagine has no significant impact on the ammonia-dependent synthetase activity at 1 mM
-
additional information
A9XS73
PvNAS2 is downregulated when carbon availability is reduced in nodules
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
glycerol
-
required for optimum activity
Phytohemagglutinin
-
-
-
mannitol
-
300 mM, induces expression of the enzyme
additional information
-
1 mM aminooxyacetic acid increases AS activity in crude extract by inhibiting various aminotransferases and preventing the assimilation of aspartate into the TCA cycle
-
additional information
-
AS expression in roots is induced by NH4+ or glutamine, but not by nitrate, glutamate, aspartate and asparagine
-
additional information
A9XS73
addition of sugars to the plants, mainly glucose, induces the enzymes leading to the increased asparagine production
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.53
Asp
-
NH4+-dependent activity, wild-type
0.6
Asp
-
-
0.68
Asp
-
wild-type enzyme
0.85
Asp
-
C436A mutant
0.85
Asp
-
Asp, wild-type enzyme, Gln-dependent activity
0.88
Asp
-
C386A mutant
0.96
Asp
-
C99A mutant
1.24
Asp
-
-
1.3
Asp
-
-
1.68
Asp
-
C168A mutant
1.7
Asp
-
C514A mutant
1.91
Asp
-
C423A mutant
55.7
Asp
-
C523A mutant
0.38
aspartic acid
-
pH 8, reaction with glutamine, C-terminally tagged recombinant enzyme
0.013
ATP
-
mutant E348D, glutamine-dependent activity, pH 8.0, 37C; mutant E348D, synthetase activity, glutamine-dependent activity, 20 mM L-Gln, pH 8.0, 37C
0.03
ATP
-
mutant E348D, ammonia-dependent activity, pH 8.0, 37C; mutant E348D, synthetase activity, ammonia-dependent activity, 100 mM NH4Cl, pH 8.0, 37C
0.08
ATP
-
pH 8, reaction with glutamine, C-terminally tagged recombinant enzyme
0.097
ATP
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
pH 7.6, 22C; pH 7.6, temperature not specified in the publication, recombinant His-tagged isozyme ZmAsn3
0.1 - 1
ATP
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
pH 7.6, temperature not specified in the publication, recombinant His-tagged isozyme ZmAsn1
0.1
ATP
-
wild-type, glutamine-dependent activity, pH 8.0, 37C; wild-type, synthetase activity, glutamine-dependent activity, 20 mM L-Gln, pH 8.0, 37C
0.106
ATP
-
pH 7.6, 22C
0.11
ATP
-
pH 8, reaction with NH3, C-terminally tagged recombinant enzyme
0.11
ATP
-
wild-type, ammonia-dependent activity, pH 8.0, 37C; wild-type, synthetase activity, ammonia-dependent activity, 100 mM NH4Cl, pH 8.0, 37C
0.11
ATP
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
pH 7.6, 22C
0.125
ATP
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
pH 7.6, 22C; pH 7.6, temperature not specified in the publication, recombinant nontagged isozyme soluble ZmAsn2
0.128
ATP
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
pH 7.6, 22C; pH 7.6, temperature not specified in the publication, recombinant His-tagged isozyme ZmAsn2; pH 7.6, temperature not specified in the publication, recombinant His-tagged isozyme ZmAsn4
0.14
ATP
-
-
0.14
ATP
-
MgATP2-
0.15
ATP
-
-
0.18
ATP
-
wild-type
0.18
ATP
-
glutamine dependent asparagine synthetase activity
0.2
ATP
-
ATP, C386A mutant
0.2
ATP
-
-
0.21
ATP
-
C99A mutant
0.24
ATP
-
C523A mutant
0.24
ATP
-
ATP
0.26
ATP
-
-
0.29
ATP
-
C436A mutant and C514A mutant
0.39
ATP
-
C168A mutant
0.44
ATP
-
C423A mutant
0.7
ATP
-
-
0.16
Gln
-
-
0.16
Gln
-
-
0.16
Gln
-
-
0.37
Gln
-
C523A mutant
0.51
Gln
-
C386A mutant
0.66
Gln
-
wild-type enzyme
0.74
Gln
-
C436 mutant
0.77
Gln
-
C423A mutant
0.83
Gln
-
C168A mutant
0.92
Gln
-
C514 mutant
1.4
Gln
-
-
5.24
Gln
-
mutant C99A
11.5
hydroxylamine
-
wild-type enzyme
17.1
hydroxylamine
-
mutant N74A
0.13
L-Asp
-
mutant E348D, glutamine-dependent activity, pH 8.0, 37C; mutant E348D, synthetase activity, glutamine-dependent activity, 20 mM L-Gln, pH 8.0, 37C
0.23
L-Asp
-
mutant E348D, ammonia-dependent activity, pH 8.0, 37C; mutant E348D, synthetase activity, ammonia-dependent activity, 100 mM NH4Cl, pH 8.0, 37C
0.31
L-Asp
-
-
0.58
L-Asp
-
wild-type, glutamine-dependent activity, pH 8.0, 37C; wild-type, synthetase activity, glutamine-dependent activity, 20 mM L-Gln, pH 8.0, 37C
0.58
L-Asp
-
-
0.76
L-Asp
-
-
0.8
L-Asp
-
-
0.85
L-Asp
-
pH 7.6, 22C
0.9
L-Asp
Q8KN11
pH 7.4, 37C
0.9
L-Asp
-
-
0.91
L-Asp
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
pH 7.6, 22C
0.93
L-Asp
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
pH 7.6, 22C
0.98
L-Asp
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
pH 7.6, 22C
1.2
L-Asp
-
wild-type, ammonia-dependent activity, pH 8.0, 37C; wild-type, synthetase activity, ammonia-dependent activity, 100 mM NH4Cl, pH 8.0, 37C
1.2
L-Asp
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
pH 7.6, 22C
0.1 - 2
L-aspartate
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
pH 7.6, temperature not specified in the publication, recombinant His-tagged isozyme ZmAsn3
0.68
L-aspartate
-
glutamine dependent asparagine synthetase activity
0.85
L-aspartate
-
-
0.9 - 1
L-aspartate
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
pH 7.6, temperature not specified in the publication, recombinant nontagged soluble isozyme ZmAsn2
0.93
L-aspartate
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
pH 7.6, temperature not specified in the publication, recombinant His-tagged isozyme ZmAsn4
0.98
L-aspartate
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
pH 7.6, temperature not specified in the publication, recombinant His-tagged isozyme ZmAsn1; pH 7.6, temperature not specified in the publication, recombinant His-tagged isozyme ZmAsn2
1.3
L-aspartic acid
-
pH 8, reaction with NH3, C-terminally tagged recombinant enzyme
0.04
L-Gln
-
mutant N74A
0.04
L-Gln
-
-
0.11
L-Gln
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
pH 7.6, 22C
0.233
L-Gln
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
pH 7.6, 22C
0.254
L-Gln
-
pH 7.6, 22C
0.423
L-Gln
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
pH 7.6, 22C
0.543
L-Gln
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
pH 7.6, 22C
0.69
L-Gln
-
wild-type enzyme
1
L-Gln
-
-
1.1
L-Gln
-
mutant E348D, glutaminase activity, pH 8.0, 37C, 5 mM ATP present
1.1
L-Gln
-
-
1.7
L-Gln
-
wild-type, glutaminase activity, pH 8.0, 37C, 5 mM ATP present
2.7
L-Gln
-
mutant E348D, glutaminase activity, pH 8.0, 37C, ATP absent
3.5
L-Gln
-
mutant E348A, glutaminase activity, pH 8.0, 37C, 5 mM ATP present
3.9
L-Gln
-
mutant E348Q, glutaminase activity, pH 8.0, 37C, 5 mM ATP present
4.3
L-Gln
-
-
5
L-Gln
-
wild-type, glutaminase activity, pH 8.0, 37C, ATP absent
5.8
L-Gln
-
mutant E348A, glutaminase activity, pH 8.0, 37C, ATP absent
9
L-Gln
-
mutant E348Q, glutaminase activity, pH 8.0, 37C, ATP absent
0.09
L-glutamic acid gamma-monohydroxamate
-
mutant N74A
0.26
L-glutamic acid gamma-monohydroxamate
-
wild-type enzyme
0.26
L-glutamic acid gamma-monohydroxamate
-
ATP, wild-type enzyme, Gln-dependent activity
0.09
L-glutamine
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
pH 7.6, temperature not specified in the publication, recombinant His-tagged isozyme ZmAsn2
0.1 - 1
L-glutamine
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
pH 7.6, temperature not specified in the publication, recombinant nontagged soluble isozyme ZmAsn2
0.233
L-glutamine
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
pH 7.6, temperature not specified in the publication, recombinant His-tagged isozyme ZmAsn4
0.423
L-glutamine
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
pH 7.6, temperature not specified in the publication, recombinant His-tagged isozyme ZmAsn3
0.543
L-glutamine
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
pH 7.6, temperature not specified in the publication, recombinant His-tagged isozyme ZmAsn1
0.69
L-glutamine
-
glutamine dependent asparagine synthetase activity
0.69
L-glutamine
-
glutamine-dependent synthetase activity
1.39
L-glutamine
-
glutaminase activity in the presence of ATP
1.67
L-glutamine
-
glutaminase activity
1.9
L-glutamine
-
glutaminase activity in the absence of ATP
1.9
L-glutamine
-
pH 8, C-terminally tagged recombinant enzyme
0.75
NH3
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
pH 7.6, 22C
1.7
NH3
-
pH 8, C-terminally tagged recombinant enzyme
8.4
NH3
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
pH 7.6, 22C
9
NH3
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
pH 7.6, 22C
9.9
NH3
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
pH 7.6, 22C
2.1
NH4+
-
-
2.7
NH4+
-
-
9
NH4+
-
-
10
NH4+
-
-
13
NH4+
-
mutant N74A
15.7
NH4+
-
wild-type enzyme
17
NH4+
-
wild-type enzyme
120
NH4+
-
-
0.076
MgATP2-
-
-
additional information
additional information
-
-
-
additional information
additional information
-
Km-values of mutant enzymes R30A, R30K, N74A, N74Q, and N79A
-
additional information
additional information
-
Km-values of a number of site-specific mutant enzymes
-
additional information
additional information
-
regulation: coregulation by light of the activities of three crucial enzymes of NH4+ assimilation and transport
-
additional information
additional information
-
kinetic mechanism, kinetic model
-
additional information
additional information
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
the AsnS isozymes are kinetically distinct with substantial differences in Km (Gln) and Vmax values, overview. None of the enzymes has cooperative enzyme kinetics; the AsnS isozymes are kinetically distinct with substantial differences in Km (Gln) and Vmax values, overview. None of the enzymes has cooperative enzyme kinetics; the AsnS isozymes are kinetically distinct with substantial differences in Km (Gln) and Vmax values, overview. None of the enzymes has cooperative enzyme kinetics; the AsnS isozymes are kinetically distinct with substantial differences in Km (Gln) and Vmax values, overview. None of the enzymes has cooperative enzyme kinetics
-
additional information
additional information
-
biphasic kinetic, linear portion near 0.5
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.52
Asp
-
C523A mutant
0.57
Asp
-
wild-type
0.77
Asp
-
C386A mutant
0.79
Asp
-
C514A mutant
1.05
Asp
-
wild-type enzyme
1.23
Asp
-
C168A mutant
1.35
Asp
-
C423A mutant
1.37
Asp
-
C99A mutant
0.43
ATP
-
mutant E348D, ammonia-dependent activity, pH 8.0, 37C; mutant E348D, synthetase activity, ammonia-dependent activity, 100 mM NH4Cl, pH 8.0, 37C
0.51
ATP
-
mutant E348D, glutamine-dependent activity, pH 8.0, 37C; mutant E348D, synthetase activity, glutamine-dependent activity, 20 mM L-Gln, pH 8.0, 37C
0.53
ATP
-
C523A mutant
0.78
ATP
-
C514A mutant
0.8
ATP
-
wild-type
0.87
ATP
-
C386A mutant
0.9
ATP
-
wild-type, glutamine-dependent activity, pH 8.0, 37C; wild-type, synthetase activity, glutamine-dependent activity, 20 mM L-Gln, pH 8.0, 37C
0.96
ATP
-
wild-type, ammonia-dependent activity, pH 8.0, 37C; wild-type, synthetase activity, ammonia-dependent activity, 100 mM NH4Cl, pH 8.0, 37C
1.1
ATP
-
wild-type
1.1
ATP
-
glutamine dependent asparagine synthetase activity
1.15
ATP
-
C168A mutant
1.17
ATP
-
C99A mutant
1.46
ATP
-
C423A mutant
1.6
ATP
-
pH 8, reaction with NH3, C-terminally tagged recombinant enzyme
1.7
ATP
-
pH 8, reaction with glutamine, C-terminally tagged recombinant enzyme
2.18
ATP
-
-
2.94
ATP
-
-
5.8
ATP
-
-
0.4
Gln
-
C523A mutant
0.59
Gln
-
wild-type
0.69
Gln
-
C386 mutant
0.73
Gln
-
C436 mutant
0.74
Gln
-
C514A mutant, Asp, C436A mutant
0.86
Gln
-
C168 mutant
1.09
Gln
-
C423A mutant
1.31
Gln
-
C99A mutant
1.7
glutamine
-
pH 8, C-terminally tagged recombinant enzyme
5.8
glutamine
-
-
1.03
hydroxylamine
-
wild-type enzyme
1.17
hydroxylamine
-
N74A mutant
0.3
L-Asp
-
mutant E348D, ammonia-dependent activity, pH 8.0, 37C; mutant E348D, synthetase activity, ammonia-dependent activity, 100 mM NH4Cl, pH 8.0, 37C
0.45
L-Asp
-
mutant E348D, glutamine-dependent activity, pH 8.0, 37C; mutant E348D, synthetase activity, glutamine-dependent activity, 20 mM L-Gln, pH 8.0, 37C
0.67
L-Asp
-
wild-type, glutamine-dependent activity, pH 8.0, 37C; wild-type, synthetase activity, glutamine-dependent activity, 20 mM L-Gln, pH 8.0, 37C
0.75
L-Asp
-
wild-type, ammonia-dependent activity, pH 8.0, 37C; wild-type, synthetase activity, ammonia-dependent activity, 100 mM NH4Cl, pH 8.0, 37C
5.8
L-asparagine
-
-
1.05
L-aspartate
-
glutamine dependent asparagine synthetase activity
1.56
L-aspartate
-
-
2.94
L-aspartate
-
-
1.3
L-aspartic acid
-
pH 8, reaction with glutamine, C-terminally tagged recombinant enzyme
1.7
L-aspartic acid
-
pH 8, reaction with NH3, C-terminally tagged recombinant enzyme
0.05
L-Gln
-
mutant N74A
1.01
L-Gln
-
wild-type enzyme
4
L-Gln
-
mutant E348Q, glutaminase activity, pH 8.0, 37C, ATP absent
4.1
L-Gln
-
mutant E348D, glutaminase activity, pH 8.0, 37C, ATP absent
4.45
L-Gln
-
mutant E348Q, glutaminase activity, pH 8.0, 37C, 5 mM ATP present
5.8
L-Gln
-
mutant E348A, glutaminase activity, pH 8.0, 37C, 5 mM ATP present
6.2
L-Gln
-
wild-type, glutaminase activity, pH 8.0, 37C, ATP absent
6.37
L-Gln
-
mutant E348A, glutaminase activity, pH 8.0, 37C, ATP absent
6.6
L-Gln
-
wild-type, glutaminase activity, pH 8.0, 37C, 5 mM ATP present
10.02
L-Gln
-
mutant E348D, glutaminase activity, pH 8.0, 37C, 5 mM ATP present
0.1
L-glutamic acid gamma-monohydroxamate
-
mutant N74A
0.15
L-glutamic acid gamma-monohydroxamate
-
wild-type
0.8
L-glutamine
-
glutaminase activity in the absence of ATP
1.01
L-glutamine
-
glutamine dependent asparagine synthetase activity
1.38
L-glutamine
-
glutaminase activity in the presence of ATP
2.73
L-glutamine
-
glutamine-dependent synthetase activity
3 - 6
L-glutamine
-
glutaminase activity; glutamine-dependent synthetase activity
3.38
L-glutamine
-
glutaminase activity
1.8
NH3
-
pH 8, C-terminally tagged recombinant enzyme
0.59
NH4+
-
wild-type
0.65
NH4+
-
mutant N74A
6.08
L-glutamine
-
glutaminase activity in the presence of ATP; glutamine dependent asparagine synthetase activity
additional information
additional information
-
turnover-numbers of mutant enzymes R30A, R30K, N74A, N74Q, and N79A
-
additional information
additional information
-
turnover-numbers of a number of site-specific AS-B mutant enzymes
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
8.7
ATP
-
wild-type, ammonia-dependent activity, pH 8.0, 37C; wild-type, synthetase activity, ammonia-dependent activity, 100 mM NH4Cl, pH 8.0, 37C
4
9
ATP
-
wild-type, glutamine-dependent activity, pH 8.0, 37C; wild-type, synthetase activity, glutamine-dependent activity, 20 mM L-Gln, pH 8.0, 37C
4
14.3
ATP
-
mutant E348D, ammonia-dependent activity, pH 8.0, 37C; mutant E348D, synthetase activity, ammonia-dependent activity, 100 mM NH4Cl, pH 8.0, 37C
4
39.2
ATP
-
mutant E348D, glutamine-dependent activity, pH 8.0, 37C; mutant E348D, synthetase activity, glutamine-dependent activity, 20 mM L-Gln, pH 8.0, 37C
4
0.63
L-Asp
-
wild-type, ammonia-dependent activity, pH 8.0, 37C; wild-type, synthetase activity, ammonia-dependent activity, 100 mM NH4Cl, pH 8.0, 37C
294
1.2
L-Asp
-
wild-type, glutamine-dependent activity, pH 8.0, 37C; wild-type, synthetase activity, glutamine-dependent activity, 20 mM L-Gln, pH 8.0, 37C
294
1.3
L-Asp
-
mutant E348D, ammonia-dependent activity, pH 8.0, 37C; mutant E348D, synthetase activity, ammonia-dependent activity, 100 mM NH4Cl, pH 8.0, 37C
294
3.4
L-Asp
-
mutant E348D, glutamine-dependent activity, pH 8.0, 37C
294
3.45
L-Asp
-
mutant E348D, synthetase activity, glutamine-dependent activity, 20 mM L-Gln, pH 8.0, 37C
294
0.45
L-Gln
-
mutant E348Q, glutaminase activity, pH 8.0, 37C, ATP absent
337
1.1
L-Gln
-
mutant E348A, glutaminase activity, pH 8.0, 37C, ATP absent
337
1.14
L-Gln
-
mutant E348Q, glutaminase activity, pH 8.0, 37C, 5 mM ATP present
337
1.19
L-Gln
-
wild-type, glutaminase activity, pH 8.0, 37C, ATP absent
337
1.51
L-Gln
-
mutant E348D, glutaminase activity, pH 8.0, 37C, ATP absent
337
1.66
L-Gln
-
mutant E348A, glutaminase activity, pH 8.0, 37C, 5 mM ATP present
337
3.88
L-Gln
-
wild-type, glutaminase activity, pH 8.0, 37C, 5 mM ATP present
337
9.1
L-Gln
-
mutant E348D, glutaminase activity, pH 8.0, 37C, 5 mM ATP present
337
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
1.1
alpha,beta-methylene-ATP
-
Kis, versus ATP
2.6
alpha,beta-methylene-ATP
-
Kis or Kii, versus Asp
3.2
alpha,beta-methylene-ATP
-
Kii, versus Gln
3.7
alpha,beta-methylene-ATP
-
Kii, versus NH3
2.5
AMP
-
-
0.91
AMP-PNP
-
-
0.023
Asn
-
Kis, versus Gln
0.25
asparagine
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
pH 7.6, 22C
18
beta-methylaspartate
-
-
0.021
diphosphate
-
Kis, versus ATP
0.65
diphosphate
-
Kii, versus Asp
0.85
diphosphate
-
Kii, versus NH3 or versus Gln; Kis, versus Asp
150
Glu
-
-
0.3
glutamate
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
pH 7.6, 22C
0.25
L-asparagine
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
pH 7.6, temperature not specified in the publication, recombinant nontagged soluble isozyme ZmAsn2
1.1
L-cysteine sulfinic acid
-
Kis, versus Asp
2.2
L-cysteine sulfinic acid
-
Kii, versus NH3
2.5
L-cysteine sulfinic acid
-
Kii, versus ATP
6.4
L-cysteine sulfinic acid
-
Kii, versus Gln
7.2
L-cysteine sulfinic acid
-
Kis, versus Gln
0.3
L-glutamate
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
pH 7.6, temperature not specified in the publication, recombinant nontagged soluble isozyme ZmAsn2
6.5
L-glutamic acid gamma-methyl ester
-
Kis, versus NH3
8.9
L-glutamic acid gamma-methyl ester
-
Kis, versus Gln
27
L-glutamic acid gamma-methyl ester
-
Kii, versus Asp
29
L-glutamic acid gamma-methyl ester
-
Kii, versus ATP
6.6
L-glutamic acid gamma-methylester
-
-
0.000285
sulfoximine adenylate
-
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
1.1
asparagine
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
pH 7.6, 22C
1.3
asparagine
-
pH 7.6, 22C
1.3
asparagine
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
pH 7.6, 22C
1.4
asparagine
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
pH 7.6, 22C
1.5
asparagine
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
pH 7.6, 22C
1.2
glutamate
-
pH 7.6, 22C
1.2
glutamate
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
pH 7.6, 22C
1.3
glutamate
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
pH 7.6, 22C; pH 7.6, 22C; pH 7.6, 22C
1.1
L-asparagine
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
pH 7.6, temperature not specified in the publication, recombinant His-tagged isozyme ZmAsn1
1.3
L-asparagine
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
pH 7.6, temperature not specified in the publication, recombinant His-tagged isozyme ZmAs4
1.4
L-asparagine
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
pH 7.6, temperature not specified in the publication, recombinant nontagged soluble isozyme ZmAsn2
1.5
L-asparagine
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
pH 7.6, temperature not specified in the publication, recombinant His-tagged isozyme ZmAsn3
1.2
L-glutamate
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
pH 7.6, temperature not specified in the publication, recombinant His-tagged isozyme ZmAsn4
1.3
L-glutamate
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
pH 7.6, temperature not specified in the publication, recombinant His-tagged isozyme ZmAsn1; pH 7.6, temperature not specified in the publication, recombinant His-tagged isozyme ZmAsn3; pH 7.6, temperature not specified in the publication, recombinant nontagged soluble isozyme ZmAsn2
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0.096
-
-
0.17
-
-
0.193
-
-
0.395
-
-
additional information
-
-
additional information
-
-
additional information
-
-
additional information
-
-
additional information
-
-
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
6 - 8.5
-
Gln-dependent activity
6.5 - 8
-
-
6.6 - 8
-
-
7.4 - 7.6
-
Gln-dependent activity
7.4
Q8KN11
-
7.5 - 8
-
-
7.6 - 8.2
-
-
7.6
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
assay at; assay at; assay at; assay at
7.8 - 8.2
-
-
7.8 - 8.5
-
broad optimum
7.9 - 8.3
-
-
8.2
-
-
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
6.5 - 8.5
-
minor variation of activity in this pH range
6.7 - 8.8
-
minor variation of activity in this pH range
6.9 - 8.3
-
80% of maximal activity at pH 6.9 and 8.3
7 - 9
-
pH 7.0: about 45% of maximal activity, pH 9.0: about 90% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
37
-
assay at
37
Q8KN11
-
37
-
assay at
pI VALUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
5.92
Q84LA5, Q93XP9
HvAS2, theoretical pI
6.14
Q84LA5, Q93XP9
HvAS1, theoretical pI
6.3 - 6.8
Q9SM55
sequence calculation
6.4
-
deduced from amino acid sequence
6.5
B3KYI2
calculated
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
-
TaASN1 is dramatically induced by salinity, osmotic stress and exogenous abscisic acid, TaASN2 transcripts are very low
Manually annotated by BRENDA team
Q2L9S5
the cell line expresses a Caenorhabditis elegans SID-1 (CeSID-1) transmembrane protein with the ability to uptake double-stranded RNA into the cells
Manually annotated by BRENDA team
-
high content of AS in grains in the middle stage of ripening, in vascular tissues
Manually annotated by BRENDA team
-
SB-P, dark-adapted
Manually annotated by BRENDA team
-
of seedlings
Manually annotated by BRENDA team
-
of seedlings
Manually annotated by BRENDA team
-
primary leaves
Manually annotated by BRENDA team
Q84LA5, Q93XP9
expression of HvAS1, higher mRNA levels in younger leaves than in older leaves, induced by dark treatment, induction seems to require a dramatic change in the C/N ratio since no diurnal variation is observed and up-regulation of transcription only occurs after 10 h of darkness
Manually annotated by BRENDA team
Q84LA5, Q93XP9
expression of HvAS2
Manually annotated by BRENDA team
-
first leaves, etiolated
Manually annotated by BRENDA team
-
high content of AS in leaf sheath at the second position from the fully expanded top leaf, the contents gradually decreases in leaf sheaths as a function of increasing age, in vascular tissues
Manually annotated by BRENDA team
-
upregulation in leaves infected by the bacterial pathogen Pseudomonas syringae, high activity in phloem cells of the main vascular bundles and in secondary veins of the leaf blade
Manually annotated by BRENDA team
-
Asn-resistant leukemia cells
Manually annotated by BRENDA team
-
high activity in fetal liver and low activity in adult liver
Manually annotated by BRENDA team
-
high expression in T-lineage and low expression in B-lineage
Manually annotated by BRENDA team
-
higher expression than in lymphoblastic leukemia cells
Manually annotated by BRENDA team
-
adult pancreas is the most active tissue found
Manually annotated by BRENDA team
Q84LA5, Q93XP9
low-level expression of HvAS1, unaffected by light
Manually annotated by BRENDA team
-
much lower expression levels than in shoots of etiolated plants, light treatment decreases expression levels
Manually annotated by BRENDA team
-
of mature plants
Manually annotated by BRENDA team
-
TaASN1 is dramatically induced by salinity, osmotic stress and exogenous abscisic acid, TaASN2 transcripts are very low
Manually annotated by BRENDA team
Q14SM9
expression after cultivation on nitrate. Expression of the gene is reduced to very low levels within days after submitting the plants to a N-free medium. The subsequent return to a complete medium, containing nitrate restores expression of all three genes. High and low expression of genes in the roots are associated with high and low ratios of Asn/Asp transported to the shoot through xylem
Manually annotated by BRENDA team
Q14SM9
expression of isoforms SAS1, SAS2 after cultivation on nitrate. Expression of the genes is reduced to very low levels within days after submitting the plants to a N-free medium. The subsequent return to a complete medium, containing nitrate restores expression of all three genes. High and low expression of genes in the roots are associated with high and low ratios of Asn/Asp transported to the shoot through xylem
Manually annotated by BRENDA team
A9XS73
expression of PvNAS2 in roots is confined to vascular bundles and meristematic tissues, while in root nodules its expression is solely localized to vascular traces and outer cortical cells encompassing the central nitrogen-fixing zone, but never detected in either infected or non-infected cells located in the central region of the nodule, high expression, confined to vascular bundles and meristematic tissues
Manually annotated by BRENDA team
-
high expression, confined to vascular bundles and meristematic tissues
Manually annotated by BRENDA team
-
detection in early stages of nodule development, no detection in nodules that have become competent for nitrogen fixation
Manually annotated by BRENDA team
Q14SM9
expression in absence of nitrate
Manually annotated by BRENDA team
Q14SM9
expression of isoforms SAS1, SAS2 in absence of nitrate
Manually annotated by BRENDA team
A9XS73
expression of PvNAS2 in roots is confined to vascular bundles and meristematic tissues, while in root nodules its expression is solely localized to vascular traces and outer cortical cells encompassing the central nitrogen-fixing zone, but never detected in either infected or non-infected cells located in the central region of the nodule, mainly induced during the early days of nitrogen fixation, confined to vascular traces and outer cortical cells. Down-regulation when carbon availability is reduced, while the addition of sugars to the plants induces expression
Manually annotated by BRENDA team
-
mainly induced during the early days of nitrogen fixation, confined to vascular traces and outer cortical cells. Down-regulation when carbon availability is reduced, while the addition of sugars to the plants induces expression
Manually annotated by BRENDA team
-
TaASN1 is dramatically induced by salinity, osmotic stress and exogenous abscisic acid, TaASN2 transcripts are very low
Manually annotated by BRENDA team
-
the gene for AS is predominantly expressed in shoots as compared to roots of etiolated plants, light treatment decreases expression levels
Manually annotated by BRENDA team
-
TaASN1 is dramatically induced by salinity, osmotic stress and exogenous abscisic acid, TaASN2 transcripts are very low
Manually annotated by BRENDA team
-
TaASN2 transcripts are very low, young, TaASN1 is dramatically induced by salinity, osmotic stress and exogenous abscisic acid
Manually annotated by BRENDA team
-
during the ripening of the spikelets AS contents increases during the first 21 days after flowering, then declines rapidly
Manually annotated by BRENDA team
Q84LA5, Q93XP9
expression of HvAS1
Manually annotated by BRENDA team
Q84LA5, Q93XP9
expression of HvAS2
Manually annotated by BRENDA team
additional information
Q9SM55
expression pattern of PVAS1, not expressed in nodules, not repressed by light
Manually annotated by BRENDA team
additional information
Q84LA5, Q93XP9
expression pattern, dark treatment of plants increases expression of HvAS1
Manually annotated by BRENDA team
additional information
Q84LA5, Q93XP9
expression pattern, no expression of HvAS2 in roots
Manually annotated by BRENDA team
additional information
-
organ and cellular localization
Manually annotated by BRENDA team
additional information
-
the three asparagine synthetase genes asnB, asnH and asnO are differentially expressed during cell growth, expression pattern
Manually annotated by BRENDA team
additional information
-
analysis of mRNA expression in 19 ovarian cancer cell lines
Manually annotated by BRENDA team
additional information
B3KYI2
isoform PvAs3 is ubiquitously expressed and not repressed by light
Manually annotated by BRENDA team
additional information
D9IXC8, E7EAQ2
comparative expression analysis of isozymes during pine development, quantitative real time-PCR expression analysis, overview. PpAS2 expression is essentially constitutive
Manually annotated by BRENDA team
additional information
D9IXC8, E7EAQ2
comparative expressionanalysis of isozymes during pine development, quantitative real time-PCR expression analysis, overview
Manually annotated by BRENDA team
additional information
-
the enzyme is ubiquitous in its organ distribution in mammals, but basal expression is relatively low in tissues other than the exocrine pancreas
Manually annotated by BRENDA team
additional information
Bacillus subtilis 168
-
the three asparagine synthetase genes asnB, asnH and asnO are differentially expressed during cell growth, expression pattern
-
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
Escherichia coli (strain K12)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
64000
-
SDS-PAGE
672914
65500
Q6HA26
SDS-PAGE
676719
70000
-
gel filtration
1681
105000
-
gel filtration
858
110000 - 120000
-
HPLC gel filtration
1674
110000
-
gel filtration
1688
160000
-
substrate-free enzyme, gel filtration
1711
230000
-
gel filtration
861
320000
-
in presence of MgCl2 and ATP association to a dimer, gel filtration
1711
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
?
-
x * 60000, SDS-PAGE
?
-
x * 66000, SDS-PAGE
?
-
x * 62666, deduced from nucleotide sequence of the asnB gene
?
-
x * 65608, SDS2, deduced from nucleotide sequence, x * 65182, SDS1, deduced from nucleotide sequence
?
Q84LA5, Q93XP9
x * 65200, HvAS2, amino acid sequence calculation
?
-
x * 65230, sequence calculation
?
Q9SM55
x * 65265, sequence calculation
?
Q84LA5, Q93XP9
x * 65500, amino acid sequence calculation, x * 65000, Western blot analysis, HvAS1
?
-
x * 65810, deduced from nucleotide sequence
?
B3KYI2
x * 64678, calculated
?
Q8KN11
x * 70000, SDS-PAGE and calculated, His-tagged protein
?
Q2L9S5
x * 62800, about, sequence calculation
?
D9IXC8, E7EAQ2
x * 66500, about, sequence calculation
?
D9IXC8, E7EAQ2
x * 66800, about, sequence calculation
dimer
-
2 * 61000, SDS-PAGE after treatment with dimethyl suberimidate
dimer
-
2 * 52500, possibly the enzyme population is heterogeneous with slight differences in subunit composition, SDS-PAGE after treatment with dimethyl suberimidate
dimer
-
probably 2 * 62000, SDS-PAGE
tetramer
-
4 * 57000, denaturing PAGE
monomer
-
1 * 52500, possibly the enzyme population is heterogeneous with slight differences in subunit composition, SDS-PAGE after treatment with dimethyl suberimidate
additional information
-
dimerization of the 160000 MW enzyme is induced by MgCl2 and ATP
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
Q84LA5, Q93XP9
no posttranslational regulation of HvAS1
additional information
-
not stained for carbohydrate by the periodic acid-Schiff procedure
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
AS-B is dialyzed against 20 mM Bis-Tris, pH 6.5, 150 mM NaCl, 0.5 mM EDTA, and 2 mM dithiothreitol, crystals are grown by hanging drop vapor diffusion at 4C, AS-B solution is adjusted to 6.5 mg/ml and contains 10 mM MgCl2, 5 mM glutamine, and 10 mM AMP, 0.01 ml of protein solution are mixed with 0.01 ml precipitant solution containing 14% polyethylene glycol 8000 and 50 mM HEPES, pH 7.0, these droplets are suspended against wells containing 15% polyethylene glycol 8000 and 50 mM HEPES, pH 7.0, crystals diffract to 2.0 A
-
X-ray crystal structure of AS-B complexed with glutamine and AMP
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
8
-
most stable around pH 8 at 4C
1688
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
37
-
half-life: 90 s
1711
40
-
the expression is dramatically decreased at 40C
676655
40
Q6HA26
the expression is dramatically decreased at 40C
676719
45
-
5 min, unstable
1681
56
-
half-life: 30 s
1711
additional information
-
a combination of L-Asp, ATP, and Mg2+ protects against heat inactivation
1688
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
unstable enzyme, 20-25% v/v glycerol and 65-70 mM 2-mercaptoethanol stabilize during storage at low temperatures
-
ATP protects from inactivation by UV irradiation in the presence of 8-N3ATP
-
glycerol and 2-mercaptoethanol are essential for enzyme stability during purification at 4C
-
unstable enzyme, 20-25% v/v glycerol and 65-70 mM 2-mercaptoethanol stabilize during storage at low temperatures
-
enzyme requires protection by high levels of thiols, glycerol and substrates also stabilize
-
activity is markedly decreased by freezing for 7 days at -87C in the presence of 1 mM dithiothreitol. Protection by 10 mM dithiothreitol
-
unstable enzyme, 20-25% v/v glycerol and 65-70 mM 2-mercaptoethanol stabilize during storage at low temperatures
-
ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SDS
Q6HA26
proteins are always extracted in buffer with 2% SDS, otherwise a high degree of degradation is detected
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-80C, recombinant C-terminally tagged enzyme is stable on prolonged storage
-
-25C, semicrude extract, 9-10 days, 50% loss of activity
-
-80C, semicrude extract, over 3 weeks, 30% loss of activity
-
-25C, 60% loss of activity after 3 months
-
-20C, inactivated after 3 months, reactivation by addition of dithiothreitol
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
immunoaffinity purification
-
partial
-
using Ni-NTA chromatography
-
one-step immunoaffinity purification
-
partial
-
partial
-
recombinant fusion protein consisting of the 42 kDa N-terminal region of AS and a 17 kDa tagged-region from pET32a(+) expression plasmid
-
recombinant
-
recombinant protein
Q8KN11
development of a method to refold recombinant AsnS from inclusion bodies; refolded, solubilized recombinant His-tagged isozyme AsnS1 by nickel affinity chromatography; refolded, solubilized recombinant His-tagged isozyme AsnS2 by nickel affinity chromatography, recombinant nontagged isozyme ZmAsnS2 from Escherichia coli by anion exchange chromatography; refolded, solubilized recombinant His-tagged isozyme AsnS4 by nickel affinity chromatography; refolded, solubilized recombinant His-tagged isozyme by nickel affinity chromatography
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
three asparagine snythetase genes asnB, asnH and asnO, expression in Bacillus subtilis and in Escherichia coli ME6279, sequencing, genetic organization
-
from BmN4 cells, recombinant expression of the GFP-tagged or Venus fluorescent protein-tagged enzyme in BmN4 cells
Q2L9S5
gene CaAS1, enzyme overexpression in transgenic Arabidopsis thaliana plants leads to enhanced resistance of the plants to Pseudomonas syringae pv. tomato DC3000 and Hyaloperonospora arabidopsidis, phenotypes, overview
-
and overexpression
-
cloned into a temperature-sensitive low copy plasmid, pOU71
-
expressed in Escherichia coli as a His-tagged fusion protein
-
overexpression in Escherichia col
-
cDNA, from cell culture SB-P, sequencing
-
expression in Escherichia coli
-
C-terminally tagged enzyme, baculovirus-based expression system, the recombinant enzyme is correctly processed, exhibits high activity and is stable on prolonged storage at -80C
-
cloned into a 2 m plasmid, pBS24.1GAS, suitable for replication in a Saccharomyces cerevisiae ciro strain AB116
-
expression of AS-GFP fusion protein in MOLT-4 cells
-
expression of several mutant enymes in Saccharomyces cerevisiae
-
mutant enzyme in which the N-terminal Cys is replaced by Ala is expressed in Saccharomyces
-
HvAS1, from leaves, located to the long arm of chromosome 5, sequencing; HvAS2, from leaves, located to the short arm of chromosome 3, sequencing
Q84LA5, Q93XP9
two cDNA clones LJAS1 and LJAS2, encoding different asparagine synthetases
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a fusion protein consisting of the 42 kDa N-terminal region of AS and a 17 kDa tagged-region from pET32a(+) expression plasmid, expression in Escherichia coli
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cloning of cDNA
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gene NAS2, DNA and mino acid sequence determination and analysis, promoter determination, tissue expression analysis, overview
A9XS73
PVAS1 gene, sequencing, expression in Escherichia coli
Q9SM55
PVAS2 is cloned into the expression vector pGEXKG and overexpressed as a glutathione S-transferase fusion protein. PVAS2 is cloned into pUC18, expression in Escherichia coli asparagine-auxotroph ER4813
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gene PpAS1, DNA and amino acid sequence analysis, sequence comparisons, phylogenetic analysis, genetic regulation of AS1 transcription, overview; gene PpAS2, DNA and amino acid sequence analysis, sequence comparisons, phylogenetic analysis
D9IXC8, E7EAQ2
expressed in Escherichia coli
Q6HA26
cloning and specificity of the amino acid-dependent contol of its mRNA content
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two allelic genes ASN1 and ASN2
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expression in Escherichia coli
Q8KN11
TaSN1 and TaSN2, expression in Escherichia coli; TaSN1, expression in Escherichia coli
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expression in Escherichia coli
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expression in Escherichia coli; expression in Escherichia coli; expression in Escherichia coli; expression in Escherichia coli; gene asnS2, expression of nontagged isozyme ZmAsnS2 in Escherichia coli, expression of C-terminally His-tagged isozyme AsnS2 in Escherichia coli strains BL21(DE3) and Rosetta(DE3) mainly in the inclusion bodies although small amounts of ZmAsnS2 are recovered in the soluble fraction; gene asnS3, expression of C-terminally His-tagged isozyme AsnS3 in Escherichia coli strains BL21(DE3) and Rosetta(DE3) mainly in the inclusion bodies; gene asnS4, expression of C-terminally His-tagged isozyme AsnS4 in Escherichia coli strains BL21(DE3) and Rosetta(DE3) mainly in the inclusion bodies; gene asnS, expression of C-terminally His-tagged isozyme AsnS1 in Escherichia coli strains BL21(DE3) and Rosetta(DE3) mainly in the inclusion bodies
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
infection with Xanthomonas campestris pv. vesicatoria induces early and strong CaAS1 expression in leaves. Significant induction of CaAS1 expression occurs in pepper leaves following treatment with salicylic acid, methyl jasmonate or wounding
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infection with Xanthomonas campestris pv. vesicatoria induces early and strong CaAS1 expression in pepper leaves
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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. 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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gene PpAS1 expression is upregulated during drought, and in the dark in all organs. PpAS1 transcripts accumulate with seedlings that are grown under high concentrations of nitrate or ammonium. Ability of PtMYB1 to activate the transcription of PpAS1 in vivo, PtMYB1 negatively regulates PpAS1 promoter in pine protoplasts
D9IXC8, E7EAQ2
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
C1A
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replacement of Cys1 by either Ala or Ser results in a loss of glutaminase activity and Gln-dependent activity, without any significant effect upon NH4+-dependent Asn synthesis. Kinetic parameters of the NH4+-dependent activity of H29A and H80A are unchanged with respect to wild-type AS-B, the apparent Km for Gln is increased by a factor of 4.5 in Gln-dependent Asn synthesis
C1S
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replacement of Cys1 by either Ala or Ser results in a loss of glutaminase activity and Gln-dependent activity, without any significant effect upon NH4+-dependent Asn synthesis. Kinetic parameters of the NH4+-dependent activity of H29A and H80A are unchanged with respect to wild-type AS-B, the apparent Km for Gln is increased by a factor of 4.5 in Gln-dependent Asn synthesis
E317A
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diminished glutaminase activity
E348A
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mutant exhibits similar glutaminase activity to the wild-type enzyme in the absence of ATP, but is not capable of catalyzing asparagine formation when glutamine is employed as a nitrogen source, mutant shows similar glutaminase activity to the wild-type enzyme in the absence of ATP, mutant is not capable of catalyzing asparagine formation when glutamine is employed as a nitrogen source, mutant shows no synthetase activity, ATP-dependent stimulation of glutaminase activity is less than that of wild-type enzyme
E348D
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mutant forms two molecules of diphosphate per asparagine
E348D
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mutant exhibits similar glutaminase activity to the wild-type enzyme in the absence of ATP and is capable of catalyzing asparagine formation when glutamine is employed as a nitrogen source. Formation of the beta-aspartyl-AMP intermediate, and therefore the eventual production of asparagine, is dependent on the presence of a carboxylate side chain at this position in the synthetase active site. In addition, E348 may also play a role in mediating the conformational changes needed to coordinate, albeit weakly, the glutaminase and synthetase activities of the enzyme and to establish the structural integrity of the intramolecular tunnel along which ammonia is translocated, mutant shows similar glutaminase activity to the wild-type enzyme in the absence of ATP, mutant is capable of catalyzing asparagine formation when glutamine is employed as a nitrogen source, glutaminase activity of the E348D mutant is stimulated more than 6fold relative to wild-type by the presence of 5 mM ATP, a 5fold decrease in the Km value for aspartate is observed for both the glutamine- and ammonia-dependent synthetase reactions, together with a kcat that is half of that observed for the wildtype enzyme
E348Q
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mutant exhibits similar glutaminase activity to the wild-type enzyme in the absence of ATP, but is not capable of catalyzing asparagine formation when glutamine is employed as a nitrogen source, mutant shows similar glutaminase activity to the wild-type enzyme in the absence of ATP, mutant is not capable of catalyzing asparagine formation when glutamine is employed as a nitrogen source, mutant shows no synthetase activity, ATP-dependent stimulation of glutaminase activity is less than that of wild-type enzyme
N74A
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mutant N74A, in which Asn is replaced by Ala can also use NH2OH as an alternative substrate to NH4+ and catalyze the hydrolysis of L-glutamic acid gamma-monohydroxamate
N74D
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replacement of the catalytically important residue Asn-74 by Asp, N74D, in the N-terminal domain of Escherichia coli Asn synthetase B confers nitrile hydratase activity upon the mutant enzyme. While wild-type As-B can efficiently catalyze the hydrolysis of Gln to Glu, the N74D As-B mutant exhibits very low glutaminase activity
N74X
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overexpression of a series of mutant enzymes. Site-directed mutagenesis of Asn74 shows that this residue plays a role in catalysis of nitrogen transfer from Gln. Replacement of Arg-30 by an Ala residue yields a mutant enzyme for which the apparent Km for Gln is increased in the Gln-dependent synthesis of Asn. In addition ATP-dependent stimulation of the glutaminase activity is modified or completely eliminated when Arg-30 is replaced by other amino acids
R30A
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overexpression of a series of mutant enzymes. Site-directed mutagenesis of Asn74 shows that this residue plays a role in catalysis of nitrogen transfer from Gln. Replacement of Arg-30 by an Ala residue yields a mutant enzyme for which the apparent Km for Gln is increased in the Gln-dependent synthesis of Asn. In addition ATP-dependent stimulation of the glutaminase activity is modified or completely eliminated when Arg-30 is replaced by other amino acids
R322Q
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60000fold decrease in kcat/KM for ATP-diphsphate exchange
R325A
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a number of site-specific AS-B mutants. When Arg325 is replaced by Ala or Lys, the resulting mutant enzymes possess no detectable Asn synthetase activity. Mutation of Thr322 and Thr323 also produce enzymes with altered kinetic properties, suggesting that these Thr are involved in Asp binding and/or stabilization of intermediates en route to beta-aspartyl-AMP
R325A
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no asparagine synthetase activity, glutaminase activity is retained
R325K
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no asparagine synthetase activity, glutaminase activity is retained
R325L
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a number of site-specific AS-B mutants. When Arg325 is replaced by Ala or Lys, the resulting mutant enzymes possess no detectable Asn synthetase activity. Mutation of Thr322 and Thr323 also produce enzymes with altered kinetic properties, suggesting that these Thr are involved in Asp binding and/or stabilization of intermediates en route to beta-aspartyl-AMP
T322X
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a number of site-specific AS-B mutants. When Arg325 is replaced by Ala or Lys, the resulting mutant enzymes possess no detectable Asn synthetase activity. Mutation of Thr322 and Thr323 also produce enzymes with altered kinetic properties, suggesting that these Thr are involved in Asp binding and/or stabilization of intermediates en route to beta-aspartyl-AMP
T323X
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a number of site-specific AS-B mutants. When Arg325 is replaced by Ala or Lys, the resulting mutant enzymes possess no detectable Asn synthetase activity. Mutation of Thr322 and Thr323 also produce enzymes with altered kinetic properties, suggesting that these Thr are involved in Asp binding and/or stabilization of intermediates en route to beta-aspartyl-AMP
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
additional information
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various mutant strains, deletion of asnO or asnH, singly or in combination, has no effect on growth rates in media with or without asparagine, deletion of asnB leads to a slow-growth phenotype, even in the presence of asparagine, strains lacking asnO cannot sporulate
additional information
Bacillus subtilis 168
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various mutant strains, deletion of asnO or asnH, singly or in combination, has no effect on growth rates in media with or without asparagine, deletion of asnB leads to a slow-growth phenotype, even in the presence of asparagine, strains lacking asnO cannot sporulate
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additional information
Q2L9S5
RNA interference is used to silence BmASNS expression in silkworm cells
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
additional information
B3KYI2
PvAs3 is able to complement Escherichia coli asparagine auxotroph strain ER
additional information
D9IXC8, E7EAQ2
PpAS2 transcript abundance is not affected by any nitrogen treatments or by water stress, PpAS2 expression is essentially constitutive
up
D9IXC8, E7EAQ2
gene PpAS2 expression is upregulated during drought, but the level of PpAS2 transcripts is not altered by darkness
additional information
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design of a silencing construct that simultaneously targets the expression of both isoforms StAs1and StAs2. Tubers of the transformed intragenic plants contain up to 20fold reduced levels of free asparagine. This coincides with a small increase in the formation of glutamine and does not affect tuber shape or yield. Heat-processed products derived from the low-asparagine tubers are indistinguishable from their untransformed counterparts in terms of sensory characteristics. However, both French fries and potato chips accumulate as little as 5% of the acrylamide present in wild-type controls
additional information
Q8KN11
after ste10 gene knock-out, the monosaccharide composition of the exopolysaccharide produced by the mutant is changed in comparison with that of native exopolysaccharide Ebosin while its antagonist activity for IL-1R decreases significantly
Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
refolding of recombinant C-terminally His-tagged enzyme from Escherichia coli strains BL21(DE3) and Rosetta(DE3) inclusion bodies to active proteins, method development using 6-8 M guanidine-HCl, overview. Vmax of the enzyme is lowered about 30% by refolding, but Km values for all three substrates show no substantial change; refolding of recombinant C-terminally His-tagged enzyme from Escherichia coli strains BL21(DE3) and Rosetta(DE3) inclusion bodies to active proteins, method development using 6-8 M guanidine-HCl, overview. Vmax of the enzyme is lowered about 30% by refolding, but Km values for all three substrates show no substantial change; refolding of recombinant C-terminally His-tagged enzymes from Escherichia coli strains BL21(DE3) and Rosetta(DE3) inclusion bodies to active proteins, method development using 6-8 M guanidine-HCl, overview. Vmax of the enzyme is lowered about 30% by refolding, but Km values for all three substrates show no substantial change; refolding of recombinant C-terminally His-tagged isozyme AsnS1 from Escherichia coli strains BL21(DE3) and Rosetta(DE3) inclusion bodies to active protein, method development using 6-8 M guanidine-HCl, overview. Vmax of the enzyme is lowered about 30% by refolding, but Km values for all three substrates show no substantial change
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
analysis
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high-performance liquid chromatography assay for Asn synthetase is an extremly sensitive and reliable method for assaying Asn synthetase
agriculture
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transgenic Arabidopsis thaliana plants that overexpress CaAS1 exhibit enhanced resistance to Pseudomonas syringae pv. tomato DC3000 and Hyaloperonospora arabidopsidis. Increased CaAS1 expression influences early defense responses in diseased leaves, including increased electrolyte leakage, reactive oxygen species and nitric oxide burst. In CaAS1-silenced pepper and/or CaAS1-overexpressing Arabidopsis, CaAS1-dependent changes in asparagine levels correlate with increased susceptibility or defense responses to microbial pathogens, respectively
analysis
P08243
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
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analysis of asparagine synthetase mRNA expression in 19 ovarian cancer cell lines. L-Asparaginase activity is only weakly correlated with asparagine synthetase mRNA expression, while asparagine synthetase protein expression measured by liquid-phase immunoassay exhibits a much stronger correlation
medicine
P08243
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
analysis
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a rapid, inexpensive micro assay that can be adapted for large numbers of samples
nutrition
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design of a silencing construct that simultaneously targets the expression of both isoforms StAs1and StAs2. Tubers of the transformed intragenic plants contain up to 20fold reduced levels of free asparagine. This coincides with a small increase in the formation of glutamine and does not affect tuber shape or yield. Heat-processed products derived from the low-asparagine tubers are indistinguishable from their untransformed counterparts in terms of sensory characteristics. However, both French fries and potato chips accumulate as little as 5% of the acrylamide present in wild-type controls
analysis
B4FFJ0, B5U8J7, B5U8J8, B5U8J9
development of a sensitive, non-radioactive assay for AsnS, based on incubation of desalted enzyme and substrates and then direct detection of either product asparagine or glutamate by HPLC