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

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

EC NUMBER
COMMENTARY
6.3.5.2
-
RECOMMENDED NAME
GeneOntology No.
GMP synthase (glutamine-hydrolysing)
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
ATP + XMP + L-glutamine + H2O = AMP + diphosphate + GMP + L-glutamate
show the reaction diagram
ATP + xanthosine 5'-phosphate + NH3 = AMP + phosphate + GMP; overall reaction
-
-
-
ATP + XMP + L-glutamine + H2O = AMP + diphosphate + GMP + L-glutamate
show the reaction diagram
steady-state ordered binding of ATP followed by XMP to the ATPase domain with glutamine binding in a random manner to the GAT domain
-
ATP + XMP + L-glutamine + H2O = AMP + diphosphate + GMP + L-glutamate
show the reaction diagram
evidence for ammonia channeling. Ammonia released from glutamine is not equilibrated with the external medium. Saturating concentrations of glutamine do not obliterate the incorporation of external ammonia into GMP. Ammonia in the external medium can access the thioester intermediate when the ATPPase domain is bound to substrates
-
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 ACCESSION NO.
COMMENTARY
LITERATURE
amination
-
-
-
-
PATHWAY
KEGG Link
MetaCyc Link
Drug metabolism - other enzymes
-
guanosine ribonucleotides de novo biosynthesis
-
Metabolic pathways
-
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.
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
di-GMP synthase A
-
-
GATase
O59071
GMP synthase [glutamine-hydrolyzing] subunit A, glutamine amidotransferase activity
GATase
Pyrococcus horikoshii OT-3
O59071
GMP synthase [glutamine-hydrolyzing] subunit A, glutamine amidotransferase activity
-
Glutamine amidotransferase
-
-
-
-
GMP synthase
-
-
-
-
GMP synthase [glutamine-hydrolyzing] subunit B
O59072
ATPPase, ATP diphosphatase activity
GMP synthase [glutamine-hydrolyzing] subunit B
Pyrococcus horikoshii OT-3
O59072
ATPPase, ATP diphosphatase activity
-
GMP synthetase
-
-
-
-
GMP synthetase
-
-
GMP synthetase
-
-
GMP synthetase
Trichomonas vaginalis 3001
-
-
-
GMP synthetase (glutamine hydrolysing)
-
-
-
-
GMPS
-
-
-
-
GMPS
-
-
GMPS
Pyrococcus horikoshii OT-3
O59072
-
-
Guanosine 5'-monophosphate synthetase
-
-
-
-
Guanosine 5'-monophosphate synthetase
-
-
Guanosine 5'-monophosphate synthetase
O59072
-
Guanosine 5'-monophosphate synthetase
Pyrococcus horikoshii OT-3
O59072
-
-
Guanosine 5-monophosphate synthetase
-
-
-
-
Guanosine 5-monophosphate synthetase
-
-
guanosine monophosphate synthetase
-
-
Guanosine monophosphate synthetase (glutamine-hydrolyzing)
-
-
-
-
Guanylate synthetase
-
-
-
-
PD0279
-
gene name
PH1346
O59071
ordered locus name
PH1346
Pyrococcus horikoshii OT-3
O59071
ordered locus name
-
Synthetase, guanylate
-
-
-
-
two-subunit-type GMP synthetase
O59072
-
two-subunit-type GMP synthetase
Pyrococcus horikoshii OT-3
O59072
-
-
Xanthosine 5'-phosphate amidotransferase
-
-
-
-
Xanthosine 5-monophosphate aminase
-
-
-
-
Xanthosine-5'-phosphate-ammonia ligase
-
-
-
-
Xanthosine-5'-phosphate:ammonia ligase (AMP-forming)
-
-
-
-
XMP aminase
-
-
-
-
Guanylate synthetase (glutamine-hydrolyzing)
-
-
-
-
additional information
O59072
GMPS belongs to the class I glutamine amidotransferase, GATase, family
additional information
Pyrococcus horikoshii OT-3
O59072
GMPS belongs to the class I glutamine amidotransferase, GATase, family
-
CAS REGISTRY NUMBER
COMMENTARY
37318-71-1
not distinguished from EC 6.3.4.1
9023-55-6
EC 6.3.4.1
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
1411, wild type and mutant enzymes
-
-
Manually annotated by BRENDA team
wild type strain AG169, psicofuranine-resistant mutant GP-1, and decoyinine-resistant mutants MG-1 and MG-4
-
-
Manually annotated by BRENDA team
strain CAI4, gene GUA1
-
-
Manually annotated by BRENDA team
Candida albicans CAI4
strain CAI4, gene GUA1
-
-
Manually annotated by BRENDA team
B; strain B96
-
-
Manually annotated by BRENDA team
Escherichia coli B96
strain B96
-
-
Manually annotated by BRENDA team
Escherichia coli K12
K12
-
-
Manually annotated by BRENDA team
with diagnosed metastatic neuroblastoma
-
-
Manually annotated by BRENDA team
strain X202
SwissProt
Manually annotated by BRENDA team
Lactobacillus rhamnosus X202
strain X202
SwissProt
Manually annotated by BRENDA team
Methanothermobacter thermautotrophicus 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
GMP synthase [glutamine-hydrolyzing] subunit A
UniProt
Manually annotated by BRENDA team
GMP synthase [glutamine-hydrolyzing] subunit B; strain OT3
UniProt
Manually annotated by BRENDA team
Pyrococcus horikoshii OT-3
-
UniProt
Manually annotated by BRENDA team
Pyrococcus horikoshii OT-3
GMP synthase [glutamine-hydrolyzing] subunit A
UniProt
Manually annotated by BRENDA team
Pyrococcus horikoshii OT-3
GMP synthase [glutamine-hydrolyzing] subunit B; strain OT3
UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
malfunction
-
GMPS/USP7 complex mutants display severe misregulation of ecdysone target genes
malfunction
-
cgsA mutants exhibit a hyperadhesive phenotype in vitro and overexpress gumJ, hxfA, hxfB, xadA, and fimA, which promote attachment of cells to surfaces and biofilm formation. The mutants are greatly reduced in virulence to grape albeit still transmissible by insect vectors, although at a reduced level compared with transmission rates of the wild-type strain, despite the fact that similar numbers of cells of the cgsA mutant are acquired by the insects from infected plants. High levels of EPS are measured in cgsA mutants compared with wild-type strains. Scanning electron microscopy analysis reveal a thicker amorphous layer surrounding the mutants. Overexpression of cgsA in a cgsA-complement mutant conferred the opposite phenotypes in vitro
metabolism
-
key enzyme in the purine salvage pathways
metabolism
O59071, O59072, -
GMPS catalyzes the final step of the de novo synthetic pathway of purine nucleotides
metabolism
Pyrococcus horikoshii OT-3
-
GMPS catalyzes the final step of the de novo synthetic pathway of purine nucleotides
-
physiological function
-
the enzymatic activities encoded by the guaAB operon are essential for Borrelia burgdorferi mouse infectivity and provide a growth advantage to spirochetes in the tick, GuaA protein is expressed during infection of mammalian hosts and is critical for the survival of Borrelia burgdorferi in the infection cycle, overview
physiological function
-
GMPS is required for its ability to deubiquitylate histone H2B. A key enzyme in the guanine nucleotide biosynthesis pathway is involved in nuclear receptor target silencing suggests that GMPS might provide a relay between metabolic state and developmental gene switching; GMP synthetase binds ubiquitin-specific protease USP7 and is required for its ability to deubiquitylate histone H2B. Strong cooperation between GMPS and USP7, which is counteracted by the histone H2B ubiquitin ligase BRE1. Loss of either GMPS or USP7 leads to increased levels of histone H2Bub in mutant animals. GMPS/USP7 binds ecdysone-regulated loci and mutants display severe misregulation of ecdysone target genes. Ecdysone receptor EcR interacts biochemically and genetically with GMPS/USP7. Analyses suggest that GMPS/USP7 acts as a transcriptional corepressor
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
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 + 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
Escherichia coli, Escherichia coli B96
-
-
-
-
ATP + 6-thioXMP + NH4+
AMP + phosphate + 6-thioGMP
show the reaction diagram
-
-
-
-
-
ATP + 6-thioXMP + NH4+
AMP + phosphate + 6-thioGMP
show the reaction diagram
Escherichia coli, Escherichia coli B96
-
-
-
-
ATP + 8-azaXMP + NH4+
AMP + phosphate + 8-azaGMP
show the reaction diagram
-
-
-
-
-
ATP + 8-azaXMP + NH4+
AMP + phosphate + 8-azaGMP
show the reaction diagram
Escherichia coli, Escherichia coli B96
-
-
-
-
ATP + beta-arabinofuranosyl-XMP
AMP + phosphate + beta-arabinofuranosylGMP
show the reaction diagram
-
-
-
-
-
ATP + dXMP + NH4+
AMP + phosphate + dGMP
show the reaction diagram
-
-
-
-
-
ATP + dXMP + NH4+
AMP + phosphate + dGMP
show the reaction diagram
-
at 4.3% of the activity relative to XMP
-
-
-
ATP + xanthosine 5'-phosphate + Gln
AMP + diphosphate + Glu
show the reaction diagram
Trichomonas vaginalis, Trichomonas vaginalis 3001
-
-
-
-
?
ATP + xanthosine 5'-phosphate + L-glutamine + H2O
AMP + diphosphate + GMP + L-glutamate
show the reaction diagram
-
-
-
?
ATP + xanthosine 5'-phosphate + L-glutamine + H2O
AMP + diphosphate + GMP + L-glutamate
show the reaction diagram
P04079
-
-
?
ATP + xanthosine 5'-phosphate + L-glutamine + H2O
AMP + diphosphate + GMP + L-glutamate
show the reaction diagram
-
GMP synthetase catalyzes the conversion of XMP to GMP. Ammonia, generated in the amino-terminal glutamine amidotransferase (GAT) domain, is transferred to the ATP-pyrophosphatase (ATPP) domain, where it attacks a highly reactive adenyl-XMP intermediate, leading to GMP formation
-
-
?
ATP + xanthosine 5'-phosphate + L-glutamine + H2O
AMP + diphosphate + GMP + L-glutamate
show the reaction diagram
-
steady state ordered binding of the substrates ATP and XMP to the ATPase domain, with ATP being the first substrate to bind followed by XMP
-
-
?
ATP + xanthosine 5'-phosphate + NH3
AMP + diphosphate + GMP
show the reaction diagram
O59071, O59072, -
overall reaction
-
-
?
ATP + xanthosine 5'-phosphate + NH3
AMP + diphosphate + GMP
show the reaction diagram
-
ATPP/DD (a construct that contains the ATP-pyrophosphatase domain as well as the predicted dimerization domain) expressed as both a His-tagged fusion protein (His-ATPP/DD) and as a non-fusion protein (NF-ATPP/DD) is capable of catalyzing the conversion of XMP to GMP using exogenously added ammonia
-
-
?
ATP + xanthosine 5'-phosphate + NH3
AMP + diphosphate + GMP
show the reaction diagram
Pyrococcus horikoshii OT-3
O59071, O59072
overall reaction
-
-
?
ATP + XMP + Gln
AMP + diphosphate + GMP + Glu
show the reaction diagram
-
-
-
-
ATP + XMP + Gln
AMP + diphosphate + GMP + Glu
show the reaction diagram
-
-
-
-
ATP + XMP + Gln
AMP + diphosphate + GMP + Glu
show the reaction diagram
-
-
-
-
ATP + XMP + Gln
AMP + diphosphate + GMP + Glu
show the reaction diagram
-
-
-
-
ATP + XMP + Gln
AMP + diphosphate + GMP + Glu
show the reaction diagram
-
-
-
-
ATP + XMP + Gln
AMP + diphosphate + GMP + Glu
show the reaction diagram
-
-
-
-
ATP + XMP + Gln
AMP + diphosphate + GMP + Glu
show the reaction diagram
Pigeon
-
ir
-
-
ATP + XMP + Gln
AMP + diphosphate + GMP + Glu
show the reaction diagram
-
maximal activity with NH4+ is 80% of that obtained with Gln
-
-
ATP + XMP + Gln
AMP + diphosphate + GMP + Glu
show the reaction diagram
-
higher Km value for NH4+ compared with Gln
-
-
ATP + XMP + Gln
AMP + diphosphate + GMP + Glu
show the reaction diagram
-
higher Km value for NH4+ compared with Gln
-
-
ATP + XMP + Gln
AMP + diphosphate + GMP + Glu
show the reaction diagram
-
higher Km value for NH4+ compared with Gln
-
-
ATP + XMP + Gln
AMP + diphosphate + GMP + Glu
show the reaction diagram
-
identical Km value for NH4+ and Gln
-
-
ATP + XMP + Gln
AMP + diphosphate + GMP + Glu
show the reaction diagram
-
identical Km value for NH4+ and Gln
-
-
ATP + XMP + Gln
AMP + diphosphate + GMP + Glu
show the reaction diagram
-
it was previously reported that the activity responsible for GMP synthesis is strictly NH4+-dependent. It is possible that the glutamine-dependent activity is inactivated by Tris in the previously used 30-min end point assay
-
-
ATP + XMP + Gln
AMP + diphosphate + GMP + Glu
show the reaction diagram
-
Gln-dependent activity is approximately 2times more active than the NH4+-dependent activity
-
-
ATP + XMP + Gln
AMP + diphosphate + GMP + Glu
show the reaction diagram
Pigeon
-
maximal rate of NH4+-dependent activity is 15% of that obtained with L-Gln
-
-
ATP + XMP + Gln
AMP + diphosphate + GMP + Glu
show the reaction diagram
-
maximal rate of the Gln-dependent activity is about 35% greater than that of NH4+-dependent activity
-
-
ATP + XMP + Gln
AMP + diphosphate + GMP + Glu
show the reaction diagram
-
a single enzyme is responsible for Gln-dependent and NH4+ dependent reaction
-
-
ATP + XMP + Gln
AMP + diphosphate + GMP + Glu
show the reaction diagram
Escherichia coli B96
-
identical Km value for NH4+ and Gln
-
-
ATP + XMP + Gln
AMP + diphosphate + GMP + Glu
show the reaction diagram
Escherichia coli B96
-
a single enzyme is responsible for Gln-dependent and NH4+ dependent reaction
-
-
ATP + XMP + Gln
AMP + diphosphate + GMP + Glu
show the reaction diagram
Escherichia coli B96
-
-
-
-
ATP + XMP + Gln
AMP + diphosphate + GMP + Glu
show the reaction diagram
Escherichia coli B96
-
identical Km value for NH4+ and Gln, Gln-dependent activity is approximately 2times more active than the NH4+-dependent activity
-
-
ATP + XMP + L-Gln
AMP + phosphate + GMP + L-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 + 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
-
-
-
-
ATP + XMP + NH4+
AMP + phosphate + GMP
show the reaction diagram
-
-
-
-
ATP + XMP + NH4+
AMP + phosphate + GMP
show the reaction diagram
-
-
-
-
ATP + XMP + NH4+
AMP + phosphate + GMP
show the reaction diagram
-
-
-
-
ATP + XMP + NH4+
AMP + phosphate + GMP
show the reaction diagram
-
-
-
-
ATP + XMP + NH4+
AMP + phosphate + GMP
show the reaction diagram
-
-
-
-
ATP + XMP + NH4+
AMP + phosphate + GMP
show the reaction diagram
-
-
-
-
ATP + XMP + NH4+
AMP + phosphate + GMP
show the reaction diagram
-
-
-
-
ATP + XMP + NH4+
AMP + phosphate + GMP
show the reaction diagram
-
-
-
-
ATP + XMP + NH4+
AMP + phosphate + GMP
show the reaction diagram
-
-
-
-
ATP + XMP + NH4+
AMP + phosphate + GMP
show the reaction diagram
-
-
-
-
ATP + XMP + NH4+
AMP + phosphate + GMP
show the reaction diagram
-
-
-
-
-
ATP + XMP + NH4+
AMP + phosphate + GMP
show the reaction diagram
-
-
-
-
ATP + XMP + NH4+
AMP + phosphate + GMP
show the reaction diagram
-
-
-
-
ATP + XMP + NH4+
AMP + phosphate + GMP
show the reaction diagram
-
substrates not mentioned:
-
-
-
ATP + XMP + NH4+
AMP + phosphate + GMP
show the reaction diagram
-
maximal activity with NH4+ is 80% of that obtained with Gln
-
-
ATP + XMP + NH4+
AMP + phosphate + GMP
show the reaction diagram
-
higher Km value for NH4+ compared with Gln
-
-
ATP + XMP + NH4+
AMP + phosphate + GMP
show the reaction diagram
-
higher Km value for NH4+ compared with Gln
-
-
ATP + XMP + NH4+
AMP + phosphate + GMP
show the reaction diagram
-
higher Km value for NH4+ compared with Gln
-
-
ATP + XMP + NH4+
AMP + phosphate + GMP
show the reaction diagram
-
identical Km value for NH4+ and Gln
-
-
ATP + XMP + NH4+
AMP + phosphate + GMP
show the reaction diagram
-
identical Km value for NH4+ and Gln
-
-
ATP + XMP + NH4+
AMP + phosphate + GMP
show the reaction diagram
-
it was previously reported that the activity responsible for GMP synthesis is strictly NH4+-dependent. It is possible that the glutamine-dependent activity is inactivated by Tris in the previously used 30-min end point assay
-
-
ATP + XMP + NH4+
AMP + phosphate + GMP
show the reaction diagram
-
Gln-dependent activity is approximately 2times more active than the NH4+-dependent activity
-
-
ATP + XMP + NH4+
AMP + phosphate + GMP
show the reaction diagram
Pigeon
-
maximal rate of NH4+-dependent activity is 15% of that obtained with L-Gln
-
-
ATP + XMP + NH4+
AMP + phosphate + GMP
show the reaction diagram
-
maximal rate of the Gln-dependent activity is about 35% greater than that of NH4+-dependent activity
-
-
ATP + XMP + NH4+
AMP + phosphate + GMP
show the reaction diagram
-
a single enzyme is responsible for Gln-dependent and NH4+ dependent reaction
-
-
ATP + XMP + NH4+
AMP + phosphate + GMP
show the reaction diagram
Escherichia coli B96
-
it was previously reported that the activity responsible for GMP synthesis is strictly NH4+-dependent. It is possible that the glutamine-dependent activity is inactivated by Tris in the previously used 30-min end point assay, maximal rate of the Gln-dependent activity is about 35% greater than that of NH4+-dependent activity
-
-
ATP + XMP + NH4+
AMP + phosphate + GMP
show the reaction diagram
Escherichia coli B96
-
identical Km value for NH4+ and Gln
-
-
ATP + XMP + NH4+
AMP + phosphate + GMP
show the reaction diagram
Escherichia coli B96
-
-
-
-
ATP + XMP + NH4+
AMP + phosphate + GMP
show the reaction diagram
Escherichia coli B96
-
-
-
-
ATP + XMP + NH4+
AMP + phosphate + GMP
show the reaction diagram
Escherichia coli B96
-
a single enzyme is responsible for Gln-dependent and NH4+ dependent reaction
-
-
ATP + XMP + NH4+
AMP + phosphate + GMP
show the reaction diagram
Escherichia coli B96
-
-
-
-
ATP + XMP + NH4+
AMP + phosphate + GMP
show the reaction diagram
Escherichia coli B96
-
-
-
-
ATP + XMP + NH4+
AMP + phosphate + GMP
show the reaction diagram
Escherichia coli B96
-
-
-
-
ATP + XMP + NH4+
AMP + phosphate + GMP
show the reaction diagram
Escherichia coli B96
-
identical Km value for NH4+ and Gln, Gln-dependent activity is approximately 2times more active than the NH4+-dependent activity
-
-
ATP + XMP + NH4+
AMP + phosphate + GMP
show the reaction diagram
Escherichia coli K12
-
-
-
-
ATPgammaS + XMP + Gln
?
show the reaction diagram
-
GMP formation is 10% that of ATP
-
-
-
L-glutamine + H2O
L-glutamate + NH3
show the reaction diagram
O59071, O59072, -
GATase half-reaction, structure-activity relationship of the functional subunits, overview
-
-
?
XMP + Mg-ATP2-
adenyl-XMP + diphosphate + Mg2+
show the reaction diagram
-
-
-
-
-
XMP + Mg-ATP2-
adenyl-XMP + diphosphate + Mg2+
show the reaction diagram
O59071, O59072, -
ATPPase half-reaction, structure-activity relationship of the functional subunits, overview
adenyl-XMP is the overall reaction intermediate
-
?
XMP + Mg-ATP2-
adenyl-XMP + diphosphate + Mg2+
show the reaction diagram
Pyrococcus horikoshii OT-3
O59071, O59072
ATPPase half-reaction, structure-activity relationship of the functional subunits, overview
adenyl-XMP is the overall reaction intermediate
-
?
L-glutamine + H2O
L-glutamate + NH3
show the reaction diagram
Pyrococcus horikoshii OT-3
O59071, O59072
GATase half-reaction, structure-activity relationship of the functional subunits, overview
-
-
?
additional information
?
-
-
maximal rate of glutaminase activity is 1.8fold greater than GMP synthetase activity, it was previously reported that the activity responsible for GMP synthesis is strictly NH4+-dependent. It is possible that the Gln-dependent activity is inactivated by Tris in the previously used 30-min end point assay
-
-
-
additional information
?
-
-
no substrates mentioned
-
-
-
additional information
?
-
-
no substrates mentioned
-
-
-
additional information
?
-
-
no substrates mentioned
-
-
-
additional information
?
-
-
no substrates mentioned
-
-
-
additional information
?
-
-
no substrates mentioned
-
-
-
additional information
?
-
-
no substrates mentioned
-
-
-
additional information
?
-
-
no substrates mentioned
-
-
-
additional information
?
-
-
glutaminase activity
-
-
-
additional information
?
-
-
glutaminase activity
-
-
-
additional information
?
-
-
final step of de novo guanine nucleotide biosynthesis
-
-
-
additional information
?
-
-
enzyme activity is linked with cellular proliferation in differentiating, regenerating and neoplastic tissues
-
-
-
additional information
?
-
-
enzyme of the purine salvage pathway
-
-
-
additional information
?
-
-
enzyme of the purine salvage pathway
-
-
-
additional information
?
-
-
key enzyme in the de novo synthesis of guanine nucleotides
-
-
-
additional information
?
-
Q9U775
GMP is necessary for DNA and RNA synthesis. Expression of the gene encoding GMP synthase coincides with the points in the life cycle with increased DNA and RNA synthesis
-
-
-
additional information
?
-
-
the enzyme is essential for de novo purine synthesis. Translocations of the MLL gene at chromosome band 11q23 are prevalent in patients with leukemia after treatment with epipodiphyllotoxins and other DNA topoisomerase II inhibitors. The MLL gene fuses with one of many different partner genes. GMPS is the first partner gene of MLL on chromosome 3Q and the first gene of this type in leukemia-associated translocations
-
-
-
additional information
?
-
-
GMP synthetase stimulates histone H2B deubiquitylation by the epigenetic silencer USP7, GMP synthetase can regulate chromatin silencing via modulation of H2B deubiquitylation by USP7
-
-
-
additional information
?
-
-
GMP synthetase binds ubiquitin-specific protease 7, USP7, and deubiquitylates histone H2B, the GMPS/USP7 complex cooperates with the Polycomb silencing system through removal of the active ubiquitin mark from histone H2B. GMPS/USP7 binds ecdysone-regulated loci prior to or after hormone signaling. The Ecdysone receptor interacts biochemically and genetically with GMPS/USP7
-
-
-
additional information
?
-
O59071, O59072, -
GMPS consists of two functional units that are present as domains or subunits: glutamine amidotransferase, GATase, and ATP pyrophosphatase, ATPPase. GATase hydrolyzes glutamine to yield glutamate and ammonia, while ATPPase utilizes ammonia to convert adenyl-XMP into guanosine 5'-monophosphate. The GATase subunit of the two-subunit-type GMPS alone is inactiv
-
-
-
additional information
?
-
Methanothermobacter thermautotrophicus Marburg
-
no substrates mentioned, enzyme of the purine salvage pathway
-
-
-
additional information
?
-
Trichomonas vaginalis 3001
-
enzyme of the purine salvage pathway
-
-
-
additional information
?
-
Escherichia coli B96
-
maximal rate of glutaminase activity is 1.8fold greater than GMP synthetase activity, it was previously reported that the activity responsible for GMP synthesis is strictly NH4+-dependent. It is possible that the Gln-dependent activity is inactivated by Tris in the previously used 30-min end point assay, glutaminase activity
-
-
-
additional information
?
-
Escherichia coli B96
-
glutaminase activity
-
-
-
additional information
?
-
Pyrococcus horikoshii OT-3
O59071, O59072
GMPS consists of two functional units that are present as domains or subunits: glutamine amidotransferase, GATase, and ATP pyrophosphatase, ATPPase. GATase hydrolyzes glutamine to yield glutamate and ammonia, while ATPPase utilizes ammonia to convert adenyl-XMP into guanosine 5'-monophosphate. The GATase subunit of the two-subunit-type GMPS alone is inactiv
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
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 + xanthosine 5'-phosphate + L-glutamine + H2O
AMP + diphosphate + GMP + L-glutamate
show the reaction diagram
P04079
-
-
?
additional information
?
-
-
final step of de novo guanine nucleotide biosynthesis
-
-
-
additional information
?
-
-
enzyme activity is linked with cellular proliferation in differentiating, regenerating and neoplastic tissues
-
-
-
additional information
?
-
-
enzyme of the purine salvage pathway
-
-
-
additional information
?
-
-
enzyme of the purine salvage pathway
-
-
-
additional information
?
-
-
key enzyme in the de novo synthesis of guanine nucleotides
-
-
-
additional information
?
-
Q9U775
GMP is necessary for DNA and RNA synthesis. Expression of the gene encoding GMP synthase coincides with the points in the life cycle with increased DNA and RNA synthesis
-
-
-
additional information
?
-
-
the enzyme is essential for de novo purine synthesis. Translocations of the MLL gene at chromosome band 11q23 are prevalent in patients with leukemia after treatment with epipodiphyllotoxins and other DNA topoisomerase II inhibitors. The MLL gene fuses with one of many different partner genes. GMPS is the first partner gene of MLL on chromosome 3Q and the first gene of this type in leukemia-associated translocations
-
-
-
additional information
?
-
-
GMP synthetase stimulates histone H2B deubiquitylation by the epigenetic silencer USP7, GMP synthetase can regulate chromatin silencing via modulation of H2B deubiquitylation by USP7
-
-
-
additional information
?
-
-
GMP synthetase binds ubiquitin-specific protease 7, USP7, and deubiquitylates histone H2B, the GMPS/USP7 complex cooperates with the Polycomb silencing system through removal of the active ubiquitin mark from histone H2B. GMPS/USP7 binds ecdysone-regulated loci prior to or after hormone signaling. The Ecdysone receptor interacts biochemically and genetically with GMPS/USP7
-
-
-
additional information
?
-
Methanothermobacter thermautotrophicus Marburg
-
enzyme of the purine salvage pathway
-
-
-
additional information
?
-
Trichomonas vaginalis 3001
-
enzyme of the purine salvage pathway
-
-
-
COFACTOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
ATP
O59071, O59072, -
as Mg-ATP
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Ca2+
-
replacement of Mg2+ with Mn2+ leads to 85% drop in activity
Mg2+
-
high activity only in presence of Mg2+
Mg2+
-
required
Mg2+
-
depends on MgATP2-, but additionally requires free Mg2+
Mg2+
-
required
Mg2+
-
required for activity
Mg2+
-
half-maximal activity at 2.1 mM, Hill-coefficient of 4.4; required for catalysis
Mg2+
O59071, O59072, -
as Mg-ATP
Mn2+
-
replacement of Mg2+ with Mn2+ leads to 70% drop in activity
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
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% inhibiton above 2 mM
2-mercaptoethanol
-
-
3'-dAMP
-
-
3'-deoxyadenosine
-
-
5'-Azido-5'-deoxyadenosine
-
-
5'-Deoxyadenosine
-
-
5'-Deoxyadenosine
-
-
5'-Thioethyl-5'-deoxyadenosine
-
-
6-Amino-9-D-psicofuranosylpurine
-
i.e. psicofuranine, irreversible inhibition, dependent on presence of XMP and diphosphate
6-Amino-9-D-psicofuranosylpurine
-
-
6-Amino-9-D-psicofuranosylpurine
-
strong inhibition of wild type enzyme, mutant enzyme GP-1 with decreased inhibition, complete loss of inhibition of mutant enzyme MG-1
6-Chloropurine ribonucleoside
-
-
6-diazo-5-oxo-L-norleucine
-
irreversible inhibition is dependent on presence of XMP, ATP and Mg2+
6-diazo-5-oxo-L-norleucine
-
irreversible inhibition of Gln-dependent activity
6-diazo-5-oxo-L-norleucine
-
inhibition of Gln-dependent activity and NH4+-dependent activity
6-diazo-5-oxo-L-norleucine
-
Gln-dependent activity is more sensitive than NH4+-dependent activity; inhibition of Gln-dependent activity and NH4+-dependent activity
6-diazo-5-oxo-L-norleucine
-
-
6-Methylmercaptopurine ribonucleoside
-
-
6-Methylpurine ribonucleoside
-
-
6-Methylpurine ribonucleotide
-
-
6-Thioguanine
-
0.5 mM, 27% inhibition
8-azaadenosine
-
-
8-Azaguanine
-
0.5 mM, 40% inhibition
8-Bromoadenosine
-
-
acivicin
-
selectively abolishes glutaminase activity and Gln-dependent synthetase activity. No effect on NH4+-dependent synthetase activity
Adenine 9-beta-D-arabinofuranoside 5'-triphosphate
-
-
adenine arabinoside
-
-
adenosine
-
strong inhibition of wild type enzyme, mutant GP-1 with decreased inhibition, complete loss of inhibition in mutant MG-1
adenosine
-
0.5 mM, 37% inhibition
adenosine 5'-(beta,gamma-imido)triphosphate
-
non-competitive versus XMP, competitive versus ATP
AMP
-
poor product inhibitor, competitive towards ATP
AMP
-
0.5 mM, 22% inhibitiion; 0.5 mM, 22% inhibition
AMP-PNP
-
potent inhibitor, competitive and non-competitive inhibitor with respect to ATP and XMP
ATP
Pigeon
-
above 5 mM
ATPgammaS
-
-
beta,gamma-Imidoadenosine 5-triphosphate
-
-
Bredinin 5'-phosphate
-
-
Bromopyruvate
-
-
Decoyinine
-
-
Decoyinine
-
uncompetitive towards Gln and XMP and noncompetitive towards ATP
Decoyinine
-
-
Decoyinine
-
strong inhibition of wild type enzyme, and mutant enzyme GP-1, complete loss of inhibition in mutant MG-1
diphosphate
-
-
diphosphate
-
competitive with respect to ATP
diphosphate
-
competitive with respect to ATP; kinetic parameters for the two progressively formed inhibitory complexes
diphosphate
-
-
diphosphate
-
non-competitive versus XMP, competitive versus ATP, non-competitive versus L-Gln; potent inhibitor, competitive and non-competitive inhibition with respect to ATP and XMP
GDP
-
0.5 mM, 42% inhibition
Glucofuranosyladenine
-
-
glutamic acid-gamma-methyl ester
-
competitive towards Gln
GMP
-
0.33 mM, 50% inhibition
GMP
-
competitive versus XMP, uncompetitive versus ATP, non-competitive versus L-Gln; potent inhibitor, competitive and uncompetitive inhibition with respect to XMP and ATP
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
-
p-chloromercuribenzoate
-
-
p-chloromercuribenzoate
-
-
p-chloromercuribenzoate
Pigeon
-
-
Psicofuranine
Q9U775
-
Psicofuranine
-
0.5 mM, 25% inhibition
Tubercidin
-
-
Zeatin ribonucleoside
-
-
additional information
-
not inhibitory at 0.5 mM: hypoxanthine, chloroadenosine, xanthosine, inosine, decoyinine
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
reduced sulfhydryl compounds
-
e.g. dithiothreitol, required
DTT
-
stimulates
additional information
-
the total number of sulfhydryl groups is approximately 22 per dimer
-
additional information
O59071, O59072, -
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
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
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
-
8-azaXMP
-
-
0.35
-
8-azaXMP
-
-
0.104
-
ATP
-
40C, pH 8.0, wild-type enzyme
0.131
-
ATP
-
native enzyme
0.132
-
ATP
-
-
0.161
-
ATP
-
recombinant enzyme
0.181
-
ATP
-
40C, pH 8.0, NF-ATPP/DD
0.2
-
ATP
-
40C, pH 8.0, His-ATPP/DD
0.26
-
ATP
-
37C, pH 8.5; pH 8.5, 37C
0.28
-
ATP
-
-
0.53
-
ATP
-
-
0.405
-
Gln
-
native enzyme
0.406
-
Gln
-
-
0.44
-
Gln
-
recombinant enzyme
0.68
-
Gln
-
-
0.0112
-
GMP
-
-
0.472
-
L-Gln
-
pH 8.5, 37C
0.472
-
L-glutamine
-
37C, pH 8.5
0.09
-
NH3
-
40C, pH 8.0, His-ATPP/DD
0.103
-
NH3
-
40C, pH 8.0, wild-type enzyme
0.127
-
NH3
-
40C, pH 8.0, NF-ATPP/DD
0.132
-
NH3
-
pH 7.4, 25C
5.1
-
NH4+
-
-
36
-
NH4+
-
-
0.0036
-
XMP
-
-
0.0044
-
XMP
-
-
0.0046
-
XMP
-
-
0.0168
-
XMP
-
37C, pH 8.5; pH 8.5, 37C
0.029
-
XMP
-
-
0.0356
-
XMP
-
native enzyme
0.041
-
XMP
-
40C, pH 8.0, NF-ATPP/DD
0.043
-
XMP
-
recombinant enzyme
0.058
-
XMP
-
-
0.096
-
XMP
-
40C, pH 8.0, His-ATPP/DD
0.166
-
XMP
-
40C, pH 8.0, wild-type enzyme
0.11
-
MgATP2-
-
-
additional information
-
additional information
-
binding dynamics at ecdysone target loci, overview
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.43
-
ATP
-
37C, pH 8.5
0.66
-
L-Gln
-
pH 7.4, 25C
0.933
-
NH3
-
pH 7.4, 25C
0.43
-
XMP
-
37C, pH 8.5; pH 8.5, 37C
0.43
-
L-glutamine
-
37C, pH 8.5
additional information
-
additional information
-
-
-
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
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
-
adenosine 5'-(beta,gamma-imido)triphosphate
-
substrate ATP, pH 8.5, 37C
0.611
-
adenosine 5'-(beta,gamma-imido)triphosphate
-
substrate XMP, pH 8.5, 37C
0.21
-
AMP-PNP
-
37C, pH 8.5, with ATP as variable substrate
0.611
-
AMP-PNP
-
37C, pH 8.5, with XMP as variable substrate
0.0364
-
diphosphate
-
37C, pH 8.5, with ATP as variable substrate; substrate ATP, pH 8.5, 37C
0.443
-
diphosphate
-
37C, pH 8.5, with XMP as variable substrate; substrate XMP, pH 8.5, 37C
0.933
-
diphosphate
-
substrate L-Gln, pH 8.5, 37C
933
-
diphosphate
-
37C, pH 8.5, with glutamine as variable substrate
0.038
-
GMP
-
37C, pH 8.5, with XMP as variable substrate; substrate XMP, pH 8.5, 37C
0.179
-
GMP
-
37C, pH 8.5, with ATP as variable substrate; substrate ATP, pH 8.5, 37C
0.39
-
GMP
-
37C, pH 8.5, with glutamine as variable substrate; substrate L-Gln, pH 8.5, 37C
0.000092
-
N2-hydroxyguanosine 5'-monophosphate
-
competitive vs XMP
0.74
-
oxanosine
-
-
IC50 VALUE [mM]
IC50 VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.5
-
guanosine
-
37C, pH 8.5
2.7
-
phosphate
-
37C, pH 8.5
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0.0428
-
-
-
0.22
-
-
-
2.8
-
-
-
3.74
-
-
-
9.09
-
-
-
additional information
-
-
-
additional information
-
Pigeon
-
-
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7
-
Pigeon
-
-
7.4
-
-
glutamine-dependent activity
7.6
7.8
-
-
7.6
-
-
-
8
-
O59071, O59072, -
assay at
8.3
8.5
-
-
8.3
-
-
Gln-dependent activity and NH4+-dependent activity
9.2
-
-
ammonia-dependent activity
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
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
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
50
-
O59071, O59072, -
assay at
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
-
Yoshida sarcoma ascites cells
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
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)
Plasmodium falciparum (isolate 3D7)
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
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
78000
-
-
gel filtration
83000
-
-
gel filtration
85000
-
-
gel filtration
120000
-
-
gel filtration
125000
-
-
gel filtration
126000
-
-
sedimentation equilibrium measurement
128000
-
-
gel filtration
160000
-
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 58604, calculation from nucleotide sequence
?
-
x * 76725, calculation from nucleotide sequence
?
-
x * 79600, calculation from nucleotide sequence
?
-
x * 65000, SDS-PAGE
?
Escherichia coli K12
-
x * 58604, calculation from nucleotide sequence
-
dimer
-
2 * 60000, SDS-PAGE in presence of 2-mercaptoethanol
dimer
-
2 * 63000, SDS-PAGE, sedimentation equilibrium measurement in 6.0 M guanidine hydrochloride
dimer
O59071, O59072, -
GMPS consists of two functional units that are present as domains or subunits: glutamine amidotransferase, GATase, and 34 kDa ATP pyrophosphatase, ATPPase. Domain structure and interactions, structure-activity relationship of the functional subunits, overview
dimer
Escherichia coli B96
-
2 * 60000, SDS-PAGE in presence of 2-mercaptoethanol; 2 * 63000, SDS-PAGE, sedimentation equilibrium measurement in 6.0 M guanidine hydrochloride; 2 * 63000, SDS-PAGE, sedimentation equilibrium measurement in 6.0 M guanidine hydrochloride
-
dimer
Pyrococcus horikoshii OT-3
-
GMPS consists of two functional units that are present as domains or subunits: glutamine amidotransferase, GATase, and 34 kDa ATP pyrophosphatase, ATPPase. Domain structure and interactions, structure-activity relationship of the functional subunits, overview
-
monomer
-
1 * 78000, SDS-PAGE
monomer
-
1 * 77360, density gradient centrifugation and subsequent electrospray MS analysis
additional information
O59071, O59072, -
the glutamine amidotransferase GATase subunit of the two-subunit-type GMPS alone is inactive, and substrates Mg2+, ATP and XMP of subunit ATP diphosphatase ATPPase except for ammonia are required to stabilize the active complex of ATPPase and GATase subunits
additional information
Pyrococcus horikoshii OT-3
-
the glutamine amidotransferase GATase subunit of the two-subunit-type GMPS alone is inactive, and substrates Mg2+, ATP and XMP of subunit ATP diphosphatase ATPPase except for ammonia are required to stabilize the active complex of ATPPase and GATase subunits
-
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
at 2.2 A resolution
-
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
O59071, O59072, -
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
O59071
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
thawing and refreezing does not diminish activity
-
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
O59071, O59072, -
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-70C, stable for at least 1 year
-
-80C, quite stable
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
purification of a stable heteromeric complex of ubiquitin-specific protease 7 (USP7) with guanosine 5'-monophosphate sythetase
-
a single enzyme is responsible for Gln-dependent and NH4+-dependent reaction
-
recombinant GMPS
-
-
Pigeon
-
Yoshida sarcoma ascites cells
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
gene guaA, the guaAB operon of Borrelia burgdorferi is present on plasmid cp26
-
the Ecdysone receptor interacts biochemically and genetically with GMPS/USP7, co-expression of GST-tagged GMPS and USP7 in Escherichia coli strain BL21(DE3)
-
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
-
expression in Escherichia coli
-
isolation of the gene
Q9U775
expression in Escherichia coli; expression in Escherichia coli; genes ph1346 and ph1347, genetic structure of GMPS, overview. Expressionin Escherichia coli strain BL21(DE3)
O59071, O59072, -
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
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
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
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
additional information
-
construction of disruption mutants, the heterozygote GUA1/gua1 strain is hypersensitive to 6-azauracil, a known inhibitor of the IMP dehydrogenase involved in GMP biosynthesis. In a murine model of systemic candidiasis, the virulence of the heterozygous strain is marginally attenuated, while the homozygous mutant gua1/gua1 strain is completely avirulent
additional information
Candida albicans CAI4
-
construction of disruption mutants, the heterozygote GUA1/gua1 strain is hypersensitive to 6-azauracil, a known inhibitor of the IMP dehydrogenase involved in GMP biosynthesis. In a murine model of systemic candidiasis, the virulence of the heterozygous strain is marginally attenuated, while the homozygous mutant gua1/gua1 strain is completely avirulent
-
Renatured/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
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 ACCESSION NO.
COMMENTARY
LITERATURE
synthesis
-
industrial production of 5-guanylic acid
synthesis
-
importance of guanine synthesis in immune cell function, GMP synthetase is a potential target for immunosuppressive therapy
medicine
Q9U775
the enzyme is a viable target for development of antimalarial chemotherapy