Information on EC 6.3.5.2 - GMP synthase (glutamine-hydrolysing)

New: Word Map on EC 6.3.5.2
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: Bacteria, Eukaryota, Archaea

EC NUMBER
COMMENTARY hide
6.3.5.2
-
RECOMMENDED NAME
GeneOntology No.
GMP synthase (glutamine-hydrolysing)
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
ATP + XMP + L-glutamine + H2O = AMP + diphosphate + GMP + L-glutamate
show the reaction diagram
ATP + XMP + NH3 = AMP + diphosphate + GMP
show the reaction diagram
(1b)
-
-
-
L-glutamine + H2O = L-glutamate + NH3
show the reaction diagram
(1a)
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
amination
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Drug metabolism - other enzymes
-
-
guanosine ribonucleotides de novo biosynthesis
-
-
Metabolic pathways
-
-
purine metabolism
-
-
Purine metabolism
-
-
SYSTEMATIC NAME
IUBMB Comments
Xanthosine-5'-phosphate:L-glutamine amido-ligase (AMP-forming)
Involved in the de novo biosynthesis of guanosine nucleotides. An N-terminal glutaminase domain binds L-glutamine and generates ammonia, which is transferred by a substrate-protective tunnel to the ATP-pyrophosphatase domain. The enzyme can catalyse the second reaction alone in the presence of ammonia.
CAS REGISTRY NUMBER
COMMENTARY hide
37318-71-1
not distinguished from EC 6.3.4.1
9023-55-6
EC 6.3.4.1
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
Borrelia burgdorferi
gene guaA
-
-
Manually annotated by BRENDA team
strain CAI4, gene GUA1
-
-
Manually annotated by BRENDA team
strain CAI4, gene GUA1
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
K12
-
-
Manually annotated by BRENDA team
strain X202
SwissProt
Manually annotated by BRENDA team
strain X202
SwissProt
Manually annotated by BRENDA team
Methanothermobacter thermautotrophicum
strain Marburg
-
-
Manually annotated by BRENDA team
Methanothermobacter thermautotrophicum Marburg
strain Marburg
-
-
Manually annotated by BRENDA team
wild type and guanine auxotroph mutant
-
-
Manually annotated by BRENDA team
Pigeon
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
metabolism
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
ATP + 1-ribosyl-4,6-dihydroxypyrazolo[3,4-d]-pyrimidine 5-phosphate
AMP + phosphate + 1-ribosyl-4-hydroxy-6-aminopyrazolo[3,4-d]pyrimidine 5'-phosphate
show the reaction diagram
ATP + 6-thioXMP + NH4+
AMP + phosphate + 6-thioGMP
show the reaction diagram
ATP + 8-azaXMP + NH4+
AMP + phosphate + 8-azaGMP
show the reaction diagram
ATP + beta-arabinofuranosyl-XMP
AMP + phosphate + beta-arabinofuranosylGMP
show the reaction diagram
-
-
-
-
-
ATP + dXMP + NH4+
AMP + phosphate + dGMP
show the reaction diagram
ATP + xanthosine 5'-phosphate + Gln
AMP + diphosphate + Glu
show the reaction diagram
ATP + xanthosine 5'-phosphate + L-glutamine + H2O
AMP + diphosphate + GMP + L-glutamate
show the reaction diagram
ATP + xanthosine 5'-phosphate + NH3
AMP + diphosphate + GMP
show the reaction diagram
ATP + XMP + Gln
AMP + diphosphate + GMP + Glu
show the reaction diagram
ATP + XMP + L-Gln
AMP + diphosphate + GMP + L-Glu
show the reaction diagram
-
enzyme can use both glutamine and external ammonia, the kinetic parameters kcat and Km for the two substrates are different and also exhibit a pH-dependent change
-
-
?
ATP + XMP + L-Gln
AMP + phosphate + GMP + L-Glu
show the reaction diagram
-
-
-
-
?
ATP + XMP + L-glutamine + H2O
AMP + diphosphate + GMP + L-glutamate
show the reaction diagram
ATP + XMP + NH3
AMP + diphosphate + GMP
show the reaction diagram
-
enzyme can use both glutamine and external ammonia, the kinetic parameters kcat and Km for the two substrates are different and also exhibit a pH-dependent change
-
-
?
ATP + XMP + NH4+
AMP + phosphate + GMP
show the reaction diagram
ATPgammaS + XMP + Gln
?
show the reaction diagram
-
GMP formation is 10% that of ATP
-
-
-
L-glutamine + H2O
L-glutamate + NH3
show the reaction diagram
XMP + Mg-ATP2-
adenyl-XMP + diphosphate + Mg2+
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
ATP + xanthosine 5'-phosphate + L-glutamine + H2O
AMP + diphosphate + GMP + L-glutamate
show the reaction diagram
ATP + XMP + L-glutamine + H2O
AMP + diphosphate + GMP + L-glutamate
show the reaction diagram
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ca2+
-
replacement of Mg2+ with Mn2+ leads to 85% drop in activity
Mn2+
-
replacement of Mg2+ with Mn2+ leads to 70% drop in activity
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(5R)-acivicin
-
-
(alpha-S,5S)-alpha-amino-3-chloro-4,5-dihydro-5-isoxazoleacetic acid
-
trivial name acivicin
1,N6-Ethenoadenosine
-
-
1,N6-Ethenoadenosine 5'-triphosphate
-
-
1-methyladenosine
-
-
1-Ribosyl-4,6-dihydroxypyrazolo[3,4-d]-pyrimidine 5'-phosphate
-
-
1-Ribosyl-4-hydroxy-6-aminopyrazolo[3,4-d]pyrimidine 5'-phosphate
-
-
2'-deoxyadenosine
-
-
2,6-Diaminopurine ribonucleoside
-
-
2-Amino-6-methylthiopurine ribonucleoside
-
-
2-chloroadenosine
-
-
2-fluoroadenosine
-
-
2-fluoroinosine 5'-monophosphate
-
50% inhibition above 2 mM
2-mercaptoethanol
-
-
3'-dAMP
-
-
3'-deoxyadenosine
-
-
5'-Azido-5'-deoxyadenosine
-
-
5'-deoxyadenosine
5'-thioethyl-5'-deoxyadenosine
-
-
6-Amino-9-D-psicofuranosylpurine
6-Chloropurine ribonucleoside
-
-
6-diazo-5-oxo-L-norleucine
6-Methylmercaptopurine ribonucleoside
-
-
6-Methylpurine ribonucleoside
-
-
6-Methylpurine ribonucleotide
-
-
6-Thio-GMP
-
-
6-ThioGMP
-
-
6-Thioguanine
-
0.5 mM, 27% inhibition
6-thioXMP
8-aza-GMP
-
-
8-azaadenosine
-
-
8-azaGMP
-
-
8-Azaguanine
-
0.5 mM, 40% inhibition
8-Bromoadenosine
-
-
acivicin
Adenine 9-beta-D-arabinofuranoside 5'-triphosphate
-
-
adenine arabinoside
-
-
adenosine
adenosine 5'-(beta,gamma-imido)triphosphate
-
non-competitive versus XMP, competitive versus ATP
AMP-PNP
-
potent inhibitor, competitive and non-competitive inhibitor with respect to ATP and XMP
ATPgammaS
-
-
azaserine
-
-
beta,gamma-Imidoadenosine 5-triphosphate
-
-
Bredinin 5'-phosphate
-
-
Bromopyruvate
-
-
CTP
-
-
Decoyinine
diphosphate
GDP
-
0.5 mM, 42% inhibition
Glucofuranosyladenine
-
-
glutamic acid-gamma-methyl ester
-
competitive towards Gln
GTP
-
0.5 mM, 40% inhibition
guanidine hydrochloride
-
-
guanosine
-
0.5 mM, 50% inhibition
hydroxylamine
-
-
IMP
-
0.5 mM, 10% inhibition
iodoacetamide
-
inhibition of Gln-dependent activity, no inhibition of NH4+-dependent activity
Kinetin ribonucleoside
-
-
L-2-Amino-4-oxo-5-chloropentanoic acid
-
inhibition of Gln-dependent activity and NH4+-dependent activity
mycophenolate
-
-
N2-hydroxyguanosine 5'-monophosphate
-
0.0003-0.0005 mM, 50% inhibition
N6,N6-Dimethyladenosine
-
-
N6-Allyladenosine
-
-
N6-Benzyladenosine
-
-
N6-Hydroxyaminopurine ribonucleoside
-
-
N6-isopentenyladenosine
-
-
N6-methyladenosine
-
-
Na+
-
25% loss of activity at 24 mM, 90% loss of activity at 200 mM
Nucleodide inhibitors
-
overview
-
oxanosine
-
-
p-chloromercuribenzoate
Psicofuranine
tubercidin
-
-
XMP
-
substrate inhibition above 0.2 mM
Zeatin ribonucleoside
-
-
additional information
-
not inhibitory at 0.5 mM: hypoxanthine, chloroadenosine, xanthosine, inosine, decoyinine
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
DTT
-
stimulates
reduced sulfhydryl compounds
-
e.g. dithiothreitol, required
additional information
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
5.4
(NH4)2SO4
-
37C, pH 8.5
2
1-ribosyl-4,6-dihydroxypyrazolo[3,4-d]-pyrimidine 5-phosphate
-
-
0.125
2-dXMP
-
-
0.42
6-thioXMP
-
-
0.05 - 0.35
8-azaXMP
0.027 - 0.53
ATP
0.21 - 0.68
Gln
0.0112
GMP
-
-
0.472
L-Gln
1.24
L-glutamine
-
at 40C in 50 mM Tris pH 7.5
0.11
MgATP2-
-
-
0.09 - 0.132
NH3
1 - 36
NH4+
0.0036 - 0.166
XMP
additional information
additional information
-
binding dynamics at ecdysone target loci, overview
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.43 - 2.39
ATP
0.66
L-Gln
Plasmodium falciparum
-
pH 7.4, 25C
0.43 - 3.05
L-glutamine
0.933
NH3
Plasmodium falciparum
-
pH 7.4, 25C
0.43 - 2.3
XMP
additional information
additional information
Homo sapiens
-
-
-
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.64
(5R)-acivicin
-
-
0.00041
(alpha-S,5S)-alpha-amino-3-chloro-4,5-dihydro-5-isoxazoleacetic acid
-
-
0.0052
6-diazo-5-oxo-L-norleucine
-
-
3
6-Thio-GMP
-
-
0.4
8-aza-GMP
-
-
0.21 - 0.611
adenosine 5'-(beta,gamma-imido)triphosphate
0.611
AMP-PNP
-
37C, pH 8.5, with XMP as variable substrate
21
ATP
-
at 40C in 50 mM Tris pH 7.5
0.0364 - 933
diphosphate
0.038 - 0.39
GMP
0.000092
N2-hydroxyguanosine 5'-monophosphate
-
competitive vs XMP
0.74
oxanosine
-
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.5
guanosine
Plasmodium falciparum
-
37C, pH 8.5
2.7
phosphate
Plasmodium falciparum
-
37C, pH 8.5
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.0428
-
-
2.8
-
-
3.74
-
-
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7
Pigeon
-
-
7.4
-
glutamine-dependent activity
7.6 - 7.8
-
-
7.8 - 8
-
-
8.3 - 8.5
-
-
8.3
-
Gln-dependent activity and NH4+-dependent activity
9.2
-
ammonia-dependent activity
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5 - 9
-
more than 50% of maximal activity at pH 6.5 and pH 9.0
7.4 - 8.6
-
more than 80% of maximal activity at pH 7.4 and 8.6
7.6 - 7.8
-
-
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Coxiella burnetii (strain RSA 493 / Nine Mile phase I)
Escherichia coli (strain K12)
Methanocaldococcus jannaschii (strain ATCC 43067 / DSM 2661 / JAL-1 / JCM 10045 / NBRC 100440)
Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
Thermoplasma acidophilum (strain ATCC 25905 / DSM 1728 / JCM 9062 / NBRC 15155 / AMRC-C165)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
78000
-
gel filtration
83000
-
gel filtration
85000
-
gel filtration
120000
-
gel filtration
125000
-
gel filtration
126000
-
sedimentation equilibrium measurement
128000
-
gel filtration
137000
-
gel filtration
160000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
monomer
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
at 2.2 A resolution
-
in complex with XMP, sitting drop vapor diffusion method, using 0.1 M sodium acetate trihydrate, pH 5.2, 1.6 M ammonium sulfate, and 0.2 M sodium chloride, at 25C
ATPPase subunit of the two-subunit-type GMPS, sitting drop vapor diffusion at 5C, mixing of 0.001 ml of protein solution containing 30 mg/ml protein in Tris-HCl, pH 8.0, with 0.001 ml of reservoir solution containing 30% v/v PEG 400, 100 mM Tris-HCl, pH 8.4, and 200 mM MgCl2, equilibration against 0.1 ml of reservoir solution, 3 weeks, X-ray diffraction structure determination and analysis at 1.8 A resolution; crystal structure of the ATPPase subunit of the two-subunit-type GMPS, to 1.79 A resolution. ATPPase consists of a N-domain and a C-domain and exists as a homodimer in the crystal and in solution. The N-domain contains an ATP-binding platform called P-loop, whereas the C-domain contains the xanthosine 5'-monophosphate-binding site and also contributes to homodimerization. The glutamine amidotransferase subunit of the two-subunit-type GMPS alone is inactive, and substrates Mg2+, ATP and XMP of PH-ATPPase except for ammonia are required to stabilize the active complex of ATPPase and GATase subunits
hanging-drop vapor-diffusion method at 5C, crystal structure is determined at 1.89 A resolution. Its overall structure and active site are the most similar to those of Escherichia coli guanosine 5'-monophosphate synthase and Sulfolobus solfataricus anthranilate synthase, respectively
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
all substrates of PH-ATPPase, Mg2+, ATP and XMP, except for ammonia, are required to stabilize the active complex of PH-ATPPase and PH-GATase subunits
thawing and refreezing does not diminish activity
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-70C, stable for at least 1 year
-
-80C, in 50 mM Tris pH 7.5, at least 3 months, no loss of activity
-
-80C, quite stable
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
a single enzyme is responsible for Gln-dependent and NH4+-dependent reaction
-
Ni-charged HiTrap chelating column chromatography and Superdex 200 gel filtration
purification of a stable heteromeric complex of ubiquitin-specific protease 7 (USP7) with guanosine 5'-monophosphate sythetase
-
Q-Sepharose column chromatography, butyl Sepharose column chromatography, and Superdex 200 gel filtration
-
recombinant GMPS
-
Yoshida sarcoma ascites cells
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
cloning of ATPP/DD (a construct that contains the ATP-pyrophosphatase domain as well as the predicted dimerization domain), expression and purification of the corresponding protein, as both a His-tagged fusion protein (His-ATPP/DD) and as a non-fusion protein (NF-ATPP/DD). Both His-ATPP/DD and NF-ATPP/DD are active proteins, capable of catalyzing the conversion of XMP to GMP using exogenously added ammonia
-
construction of a plasmid for high level expression of XMP aminase in Escherichia coli
-
expressed in Escherichia coli BL21(DE3) cells
-
expressed in Escherichia coli BL21(DE3) gold pRARE cells
expressed in Escherichia coli DH5alpha cells
-
expression in Escherichia coli
-
expression in Escherichia coli; expression in Escherichia coli; genes ph1346 and ph1347, genetic structure of GMPS, overview. Expressionin Escherichia coli strain BL21(DE3)
gene guaA, the guaAB operon of Borrelia burgdorferi is present on plasmid cp26
Borrelia burgdorferi
-
isolation of the gene
overexpression in Escherichia coli
-
the Ecdysone receptor interacts biochemically and genetically with GMPS/USP7, co-expression of GST-tagged GMPS and USP7 in Escherichia coli strain BL21(DE3)
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
GP-1
-
psicofuranine, decoyinine and adenosine strongly inhibit wild type enzyme. Mutant enzyme GP-1 shows decreased inhibition with adenosine and psicofuranine, but inhibition by decoyinine does not vary, complete loss of inhibition in mutant MG-1
C95A
-
mutant defective in glutamine hydrolysis. The phenotype of ectopic coexpression of USP7 witheither S242L or C95A is similar to that resulting from the coexpression of WT GMPS. Ectopic overexpression of only mutation S242L, mutation C95A, or USP7 mutation C250A has no effect on eye development
S242L
-
mutant defective in ATP hydrolysis. The phenotype of ectopic coexpression of USP7 with either S242L or C95A is similar to that resulting from the coexpression of WT GMPS. Ectopic overexpression of only mutation S242L, mutation C95A, or USP7 mutation C250A has no effect on eye development
37W
-
the mutant shows severely reduced specific activity compared to the wild type enzyme
W37A
-
the mutant shows slightly reduced specific activity compared to the wild type enzyme
C102A
-
totally devoid of glutamine-dependent activity. Kinetic constants for NH4Cl, ATP, and XMP obtained for the ammonia-dependent activity are similar to that of the wild-type enzyme
G388D
-
reduces the activity of GMP synthase Gua1 in budding yeast and the total G-nucleotide pool, leading to precipitous reductions in the GDP/GTP ratio and ATP level in vivo. G388D strongly reduces the rate of growth, impairs general protein synthesis, and derepresses translation of GCN4 mRNA, encoding a transcriptional activator of diverse amino acid biosynthetic enzymes. Although processing of pre-tRNAi Met and other tRNA precursors, and the aminoacylation of tRNAi Met are also strongly impaired in G388D cells, tRNAi Met-containing complexes with the macromolecular composition of the eIF2tRNAi Met.GTP complex and the multifactor complex required for translation initiation accumulate 10-fold in G388D cells and, to a lesser extent, in wild-type cells treated with 6-azauracil
additional information
Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
at low concentrations of the denaturants, up to 4.0 M urea and 1.2 M guanidine hydrochloride, inactivation is reversible. In 3.0-6.0 M guanidine hydrochloride the enzyme is extensively unfolded and partially, 15%, reassociable to the active form by removal of the denaturant. In 4.0 to 8.0 M urea and 1.2 to 3.0 M guanidine hydrochloride, the inactivation is irreversible due to the aggregation of the partially unfolded polypeptide chains
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
medicine
the enzyme is a viable target for development of antimalarial chemotherapy
synthesis
Show AA Sequence (12915 entries)
Longer loading times are possible. Please use the Sequence Search for a certain query.