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

New: Word Map on EC 6.3.5.4
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Mark a special word or phrase in this record:
Search Reference ID:
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)


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

EC NUMBER
COMMENTARY hide
6.3.5.4
-
RECOMMENDED NAME
GeneOntology No.
Asparagine synthase (glutamine-hydrolysing)
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
ATP + L-aspartate + L-glutamine + H2O = AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
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].
CAS REGISTRY NUMBER
COMMENTARY hide
37318-72-2
-
ORGANISM
COMMENTARY hide
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
strain 168, three glutamine-dependent AsnB-type asparagine snythetase genes: asnB, asnH and asnO, formerly yisO
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
; gene CaAS1
-
-
Manually annotated by BRENDA team
-
-
-
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
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
v. Little Marvel, dark-grown
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
isoforms StAs1 and StAs2
-
-
Manually annotated by BRENDA team
strain 139
UniProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
enzyme form TaSN1; enzyme form TaSN2
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
malfunction
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; 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; 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; 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
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
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 + L-Asn + L-Glu
show the reaction diagram
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 + NH3
AMP + diphosphate + Asn
show the reaction diagram
ATP + L-Asp + NH3
AMP + diphosphate + L-Asn
show the reaction diagram
ATP + L-aspartate + L-glutamine
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
ATP + L-aspartate + L-glutamine + H2O
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
ATP + L-aspartate + NH3
AMP + diphosphate + L-asparagine
show the reaction diagram
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
L-Glutamic acid gamma-monohydroxamate + H2O
Hydroxylamine + Glu
show the reaction diagram
-
-
-
-
-
L-glutamine
L-glutamate + NH3
show the reaction diagram
UTP + L-Asp + L-Gln
UMP + diphosphate + Asn + Glu
show the reaction diagram
-
weak activity
-
-
-
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
ATP + L-Asp + L-Gln
AMP + diphosphate + Asn + Glu
show the reaction diagram
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 + H2O
AMP + diphosphate + L-asparagine + L-glutamate
show the reaction diagram
ATP + L-aspartate + NH3
AMP + diphosphate + L-asparagine
show the reaction diagram
L-glutamine
L-glutamate + NH3
show the reaction diagram
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ca2+
-
at 16.7 mM CaCl2 31% of the activation relative to MgCl2
CN-
-
stimulates Asn synthesis and Gln hydrolysis
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
Na+
-
150 mM, induces expression of the enzyme
NO3-
-
stimulates Asn synthesis and Gln hydrolysis
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
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
3,4-Dicarboxypyridine
-
-
3,5-Dicarboxypyridine
-
-
4-Carboxypyridine
-
-
5'-O-[p-(fluorosulfonyl)benzoyl]adenosine
5-Bromo-4-oxo-L-norvaline
-
-
5-Chloro-4-oxo-L-norvaline
5-Diazo-4-oxo-L-norvaline
-
-
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
alpha,beta-methylene ATP
-
-
alpha,beta-methylene-ATP
-
-
aminomalonic acid
ammonium maleamate
-
weak
AMP-PNP
-
competitive vs. ATP, noncompetitive vs. aspartate, uncompetitive vs. glutamine
Arg
-
-
asparagine
aspartic acid analogs
-
-
ATP
-
strong inhibition above 5 mM
azaserine
beta,gamma-methylene ATP
-
-
beta-asparaginyladenylate
-
-
beta-Methyl-DL-Asn
-
-
beta-methylaspartate
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
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
glutamate
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
L-beta-Aspartate ethyl ester
-
-
L-cysteinesulfinic acid
-
-
L-glutamate
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
-
-
Methyl gamma-Gln
-
-
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-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
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
glycerol
mannitol
-
300 mM, induces expression of the enzyme
Phytohemagglutinin
-
-
-
additional information
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.53 - 55.7
Asp
0.38
aspartic acid
-
pH 8, reaction with glutamine, C-terminally tagged recombinant enzyme
0.013 - 1
ATP
0.16 - 5.24
Gln
11.5 - 17.1
hydroxylamine
0.13 - 1.2
L-Asp
0.1 - 2
L-aspartate
1.3
L-aspartic acid
-
pH 8, reaction with NH3, C-terminally tagged recombinant enzyme
0.04 - 9
L-Gln
0.09 - 0.26
L-glutamic acid gamma-monohydroxamate
0.1 - 1.9
L-glutamine
0.076
MgATP2-
-
-
0.75 - 9.9
NH3
2.1 - 120
NH4+
additional information
additional information
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.52 - 1.37
Asp
0.43 - 5.8
ATP
0.4 - 1.31
Gln
1.7 - 5.8
glutamine
1.03 - 1.17
hydroxylamine
0.3 - 0.75
L-Asp
5.8
L-asparagine
Vibrio cholerae
-
-
1.05 - 2.94
L-aspartate
1.3 - 1.7
L-aspartic acid
0.05 - 10.02
L-Gln
0.1 - 0.15
L-glutamic acid gamma-monohydroxamate
0.8 - 6.08
L-glutamine
1.8
NH3
Homo sapiens
-
pH 8, C-terminally tagged recombinant enzyme
0.59 - 0.65
NH4+
additional information
additional information
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
8.7 - 39.2
ATP
0.63 - 3.45
L-Asp
0.45 - 9.1
L-Gln
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.1 - 3.7
alpha,beta-methylene-ATP
2.5
AMP
-
-
0.91
AMP-PNP
-
-
0.023
Asn
-
Kis, versus Gln
0.25
asparagine
pH 7.6, 22C
18
beta-methylaspartate
-
-
0.021 - 0.85
diphosphate
150
Glu
-
-
0.3
glutamate
pH 7.6, 22C
0.25
L-asparagine
pH 7.6, temperature not specified in the publication, recombinant nontagged soluble isozyme ZmAsn2
1.1 - 7.2
L-cysteine sulfinic acid
0.3
L-glutamate
pH 7.6, temperature not specified in the publication, recombinant nontagged soluble isozyme ZmAsn2
6.5 - 29
L-glutamic acid gamma-methyl ester
6.6
L-glutamic acid gamma-methylester
-
-
0.000285
sulfoximine adenylate
-
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.1 - 1.5
asparagine
1.2 - 1.3
glutamate
1.1 - 1.5
L-asparagine
1.2 - 1.3
L-glutamate
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.096
-
-
0.17
-
-
0.193
-
-
0.395
-
-
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 8.5
-
Gln-dependent activity
6.5 - 8
-
-
6.6 - 8
-
-
7.4 - 7.6
-
Gln-dependent activity
7.5 - 8
-
-
7.6 - 8.2
-
-
7.6
assay at; assay at; assay at; assay at
7.8 - 8.5
-
broad optimum
7.8 - 8.2
-
-
7.9 - 8.3
-
-
8.2
-
-
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
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 hide
LITERATURE
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.92
HvAS2, theoretical pI
6.14
HvAS1, theoretical pI
6.3 - 6.8
sequence calculation
6.4
-
deduced from amino acid sequence
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
TaASN1 is dramatically induced by salinity, osmotic stress and exogenous abscisic acid; TaASN2 transcripts are very low
Manually annotated by BRENDA team
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
-
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
-
tip section
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
expression of HvAS1; expression of HvAS2
Manually annotated by BRENDA team
additional information