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Information on EC 6.3.4.2 - CTP synthase (glutamine hydrolysing)
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6.3.4.2 CTP synthase (glutamine hydrolysing)IUBMB Comments
The enzyme contains three functionally distinct sites: an allosteric GTP-binding site, a glutaminase site where glutamine hydrolysis occurs (cf. EC 3.5.1.2, glutaminase), and the active site where CTP synthesis takes place. The reaction proceeds via phosphorylation of UTP by ATP to give an activated intermediate 4-phosphoryl UTP and ADP [4,5]. Ammonia then reacts with this intermediate generating CTP and a phosphate. The enzyme can also use ammonia from the surrounding solution [3,6].
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Word Map
- 6.3.4.2
- pyrimidine
- cytosine
-
cytoophidia
- dctp
-
cyclopentenyl
- deoxycytidine
- acivicin
-
glutamine-dependent
- 3-deazauridine
-
rheumatology
- l-glutamine
-
alliance
-
carra
-
ctpas
- pyrazofurin
-
uridine-cytidine
-
rheumatologist
- 6-diazo-5-oxo-l-norleucine
- drug development
- medicine
- analysis
The enzyme appears in viruses and cellular organisms
Reaction Schemes
+
+
=
+
+
+
Synonyms
ctps, ctp synthetase, ctp synthase, ctps1, ctpsyn, ctps2, cytidine triphosphate synthetase, ctp synthetase 1, cytidine 5'-triphosphate synthase, ecctps, 
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SYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
CTP synthase
CTP synthase 1
CTP synthetase
CTPS
CTPS1
CTPS2
CTPsyn
cytidine 5'-triphosphate synthetase
cytidine triphosphate synthetase
PyrG
synthetase, cytidine triphosphate
-
-
-
-
URA7
uridine triphosphate aminase
-
-
-
-
UTP-ammonia ligase
-
-
-
-
CTP synthetase
-
-
-
-
CTPS
-
-
-
-
cytidine 5'-triphosphate synthetase
-
-
-
-
cytidine triphosphate synthetase
-
-
-
-
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REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ATP + UTP + NH3 = ADP + phosphate + CTP
(1b)
-
-
-
L-glutamine + H2O = L-glutamate + NH3
(1a)
-
-
-
ATP + UTP + L-glutamine = ADP + phosphate + CTP + L-glutamate
mechanism proceeds by formation of a phosphorylated pyrimidinone
-
ATP + UTP + L-glutamine = ADP + phosphate + CTP + L-glutamate
mechanism, the initial chemical step catalyzed by CTP synthetase is the phosphorylation of UTP by ATP to form an enzyme-bound intermediate
-
ATP + UTP + L-glutamine = ADP + phosphate + CTP + L-glutamate
overall reaction
-
-
-
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
amination
-
-
-
-
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PATHWAY SOURCE
PATHWAYS
BRENDA
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SYSTEMATIC NAME
IUBMB Comments
UTP:ammonia ligase (ADP-forming)
The enzyme contains three functionally distinct sites: an allosteric GTP-binding site, a glutaminase site where glutamine hydrolysis occurs (cf. EC 3.5.1.2, glutaminase), and the active site where CTP synthesis takes place. The reaction proceeds via phosphorylation of UTP by ATP to give an activated intermediate 4-phosphoryl UTP and ADP [4,5]. Ammonia then reacts with this intermediate generating CTP and a phosphate. The enzyme can also use ammonia from the surrounding solution [3,6].
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CAS REGISTRY NUMBER
COMMENTARY
9023-56-7
-
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SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
ATP + UTP + gamma-L-glutamylhydroxamate
ADP + phosphate + N4-hydroxy-CTP
-
-
-
?
ATP + UTP + hydroxylamine
ADP + phosphate + N4-hydroxyCTP
-
activity is about 3fold higher than activity with NH4+
-
?
ATP + UTP + L-gamma-glutamyl hydrazide
ADP + phosphate + N4-amino-CTP + L-glutamate
-
-
-
?
ATP + UTP + O-methylhydroxylamine
ADP + phosphate + N4-methoxyCTP
-
-
-
?
ATP + dUTP + NH4+
ADP + phosphate + dCTP
-
half-saturating concentration of 0.6 mM for dUTP, it is unlikely that this reaction plays a significant physiological role
-
?
ATP + UTP + glutamine
ADP + phosphate + CTP + Glu
-
maximal activity with NH4+ is at least 20% greater than with glutamine
-
-
?
ATP + UTP + glutamine
ADP + phosphate + CTP + Glu
Escherichia coli B / ATCC 11303
-
maximal activity with NH4+ is at least 20% greater than with glutamine
-
-
?
ATP + UTP + glutamine
ADP + phosphate + CTP + L-glutamate
-
the CTPS1-encoded enzyme is regulated by reversible phosphorylation at Thr455, regulation mechanisms, overview
-
-
?
ATP + UTP + glutamine
ADP + phosphate + CTP + L-glutamate
-
CTP synthetase is a cytosolic-associated glutamine amidotransferase enzyme that catalyzes the ATP-dependent transfer of the amide nitrogen from glutamine to the C-4 position of UTP to form CTP. CTP is an essential precursor of all membrane phospholipids that are synthesized via the Kennedy, i.e. CDP-choline and CDP-ethanolamine branches, and CDP-diacylglycerol pathways. The URA7-encoded CTP synthetase is responsible for the majority of the CTP made in vivo. Regulation mechanisms, detailed overview
-
-
?
ATP + UTP + L-glutamine
ADP + phosphate + CTP + L-glutamate
-
-
-
?
ATP + UTP + L-glutamine
ADP + phosphate + CTP + L-glutamate
-
-
-
?
ATP + UTP + L-glutamine
ADP + phosphate + CTP + L-glutamate
-
-
-
-
?
ATP + UTP + L-glutamine
ADP + phosphate + CTP + L-glutamate
-
-
-
?
ATP + UTP + L-glutamine
ADP + phosphate + CTP + L-glutamate
CTPS catalyses the ATP-dependent formation of CTP from UTP using either ammonia or L-glutamine as the nitrogen source
-
-
?
ATP + UTP + NH3
ADP + phosphate + CTP
CTPS catalyses the ATP-dependent formation of CTP from UTP using either ammonia or L-glutamine as the nitrogen source
-
-
?
ATP + UTP + NH3
ADP + phosphate + CTP
-
CTPS1 is involved in microtubule network formation and/or stabilzation, overview
-
-
?
ATP + UTP + NH4+
ADP + phosphate + CTP
last step in CTP biosynthesis
-
-
?
UTP + ATP + NH3
CTP + ADP + phosphate
-
the enzyme is regulated in a complex fashion, overview
-
-
?
UTP + ATP + NH3
CTP + ADP + phosphate
-
NH3 from glutamine deamination or exogenous
-
-
?
additional information
?
-
-
repression of the pyrG gene encoding cytidine triphosphate synthetase is responsive to cytidine nucleotide levels and is independent of both uridine nucleotides and PyrR-dependent attenuation
-
-
?
additional information
?
-
CTP synthetase plays a pivotal role in the synthesis of CTP and dCTP
-
-
?
additional information
?
-
-
CTP synthetase plays a pivotal role in the synthesis of CTP and dCTP
-
-
?
additional information
?
-
UTP- and CTP-binding structures, overview
-
-
?
additional information
?
-
-
UTP- and CTP-binding structures, overview
-
-
?
additional information
?
-
-
key enzyme for biosynthesis of cytosine ribonucleotides
-
-
?
additional information
?
-
-
the enzyme catalyzes the rate-limiting step in synthesis of cytosine nucleotides from both de novo and uridine-salvage pathways
-
-
?
additional information
?
-
-
rate-limiting enzyme in the synthesis of cytosine nucleotides from both de novo and uridine-salvage pathways. In human lymphoblastic leukemia cells the synthesis of CTP occurs predominantly via CTP synthetase, whereas in proliferating normal human T lymphocytes the salvage of cytidine is preferred
-
-
?
additional information
?
-
-
the enzyme is one of the key enzymes in pyrimidine nucleotide anabolic pathways. The activity of this enzyme is elevated in various malignancies including acute lymphocytic leukemia
-
-
?
additional information
?
-
CTP synthetase plays a pivotal role in the synthesis of CTP and dCTP
-
-
?
additional information
?
-
CTP synthetase plays a pivotal role in the synthesis of CTP and dCTP
-
-
?
additional information
?
-
-
CTP synthetase plays a pivotal role in the synthesis of CTP and dCTP
-
-
?
additional information
?
-
-
CTPS1 interacts with the GST-tagged peptidyl prolyl isomerase Pin1 from Xenopus laevis in a Ser575 phosphorylation-dependent manner, the CTPS1 also binds several other enzyme, e.g. alpha-tubulin, overview
-
-
?
additional information
?
-
-
CTP limitation increases expression of CTP synthase in Lactococcus lactis. At normal CTP concentrations a terminator is preferentially formed in the pyrG leader, thereby reducing expression of CTP synthase. At low CTP concentrations the RNA polymerase pauses at a stretch of C residues inthe pyrG leader, thereby allowing an antiterminator to form and transcription to proceed
-
-
?
additional information
?
-
-
in Lactococcus lactis the pyrG gene product is the only enzyme responsible for the amination of UTP to CTP
-
-
?
additional information
?
-
CTP synthetase plays a pivotal role in the synthesis of CTP and dCTP
-
-
?
additional information
?
-
CTP synthetase plays a pivotal role in the synthesis of CTP and dCTP
-
-
?
additional information
?
-
CTP synthetase plays a pivotal role in the synthesis of CTP and dCTP
-
-
?
additional information
?
-
CTP synthetase plays a pivotal role in the synthesis of CTP and dCTP
-
-
?
additional information
?
-
-
regulation of CTP synthetase activity by CTP plays an important role in the regulation of phospholipid synthesis
-
-
?
additional information
?
-
-
the enzyme is allosterically regulated by CTP product inhibition and by reversible phosphorylation
-
-
?
additional information
?
-
-
the enzyme can utilize NH3 as a substrate
-
-
?
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NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
ATP + UTP + NH4+
ADP + phosphate + CTP
last step in CTP biosynthesis
-
-
?
UTP + ATP + NH3
CTP + ADP + phosphate
-
the enzyme is regulated in a complex fashion, overview
-
-
?
ATP + UTP + glutamine
ADP + phosphate + CTP + L-glutamate
-
the CTPS1-encoded enzyme is regulated by reversible phosphorylation at Thr455, regulation mechanisms, overview
-
-
?
ATP + UTP + glutamine
ADP + phosphate + CTP + L-glutamate
-
CTP synthetase is a cytosolic-associated glutamine amidotransferase enzyme that catalyzes the ATP-dependent transfer of the amide nitrogen from glutamine to the C-4 position of UTP to form CTP. CTP is an essential precursor of all membrane phospholipids that are synthesized via the Kennedy, i.e. CDP-choline and CDP-ethanolamine branches, and CDP-diacylglycerol pathways. The URA7-encoded CTP synthetase is responsible for the majority of the CTP made in vivo. Regulation mechanisms, detailed overview
-
-
?
ATP + UTP + L-glutamine
ADP + phosphate + CTP + L-glutamate
-
-
-
-
?
ATP + UTP + L-glutamine
ADP + phosphate + CTP + L-glutamate
-
-
-
?
ATP + UTP + NH3
ADP + phosphate + CTP
-
CTPS1 is involved in microtubule network formation and/or stabilzation, overview
-
-
?
additional information
?
-
-
repression of the pyrG gene encoding cytidine triphosphate synthetase is responsive to cytidine nucleotide levels and is independent of both uridine nucleotides and PyrR-dependent attenuation
-
-
?
additional information
?
-
CTP synthetase plays a pivotal role in the synthesis of CTP and dCTP
-
-
?
additional information
?
-
-
CTP synthetase plays a pivotal role in the synthesis of CTP and dCTP
-
-
?
additional information
?
-
-
key enzyme for biosynthesis of cytosine ribonucleotides
-
-
?
additional information
?
-
-
the enzyme catalyzes the rate-limiting step in synthesis of cytosine nucleotides from both de novo and uridine-salvage pathways
-
-
?
additional information
?
-
-
rate-limiting enzyme in the synthesis of cytosine nucleotides from both de novo and uridine-salvage pathways. In human lymphoblastic leukemia cells the synthesis of CTP occurs predominantly via CTP synthetase, whereas in proliferating normal human T lymphocytes the salvage of cytidine is preferred
-
-
?
additional information
?
-
-
the enzyme is one of the key enzymes in pyrimidine nucleotide anabolic pathways. The activity of this enzyme is elevated in various malignancies including acute lymphocytic leukemia
-
-
?
additional information
?
-
CTP synthetase plays a pivotal role in the synthesis of CTP and dCTP
-
-
?
additional information
?
-
CTP synthetase plays a pivotal role in the synthesis of CTP and dCTP
-
-
?
additional information
?
-
-
CTP synthetase plays a pivotal role in the synthesis of CTP and dCTP
-
-
?
additional information
?
-
-
CTP limitation increases expression of CTP synthase in Lactococcus lactis. At normal CTP concentrations a terminator is preferentially formed in the pyrG leader, thereby reducing expression of CTP synthase. At low CTP concentrations the RNA polymerase pauses at a stretch of C residues inthe pyrG leader, thereby allowing an antiterminator to form and transcription to proceed
-
-
?
additional information
?
-
-
in Lactococcus lactis the pyrG gene product is the only enzyme responsible for the amination of UTP to CTP
-
-
?
additional information
?
-
CTP synthetase plays a pivotal role in the synthesis of CTP and dCTP
-
-
?
additional information
?
-
CTP synthetase plays a pivotal role in the synthesis of CTP and dCTP
-
-
?
additional information
?
-
CTP synthetase plays a pivotal role in the synthesis of CTP and dCTP
-
-
?
additional information
?
-
CTP synthetase plays a pivotal role in the synthesis of CTP and dCTP
-
-
?
additional information
?
-
-
regulation of CTP synthetase activity by CTP plays an important role in the regulation of phospholipid synthesis
-
-
?
additional information
?
-
-
the enzyme is allosterically regulated by CTP product inhibition and by reversible phosphorylation
-
-
?
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Mg2+
Mg2+
-
the enzyme requires more Mg2+ for full catalytic activity than required simply to complex the nucleotide substrate. Half-saturation value for Mg2+ is 2.6 mM for the reaction with NH4+ and 2.4 mM for the glutamine-dependent reaction
Mg2+
-
required, the activation curve for free Mg2+ concentration is greatly dependent on whether the concentration of ATP and UTP are near saturating
Mg2+
-
dUTP-dependent activity is dependent on Mg2+, maximal activity at 4 mM
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INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
2',3'-dialdehyde adenosine 5'-triphosphate
irreversible inhibitor of CTPS
2'-deoxy-GTP
no guanosine, kact: 1.5/sec, KA: 0.21 mM, Ki: 0.36 mM; the GTP analogue is capable of inhibiting Gln-dependent CTP formation at over 0.15 mM
6-thio-GTP
the GTP analogue is capable of inhibiting Gln-dependent CTP formation at over 0.15 mM
6-thioguanosine 5'-triphosphate
no guanosine, kact: 8.5/sec, KA: 0.035 mM, Ki: 0.27 mM
acivicin
-
irreversible inhibition by the glutamine analogue acivin. The acivicin inhibition of Trypanosoma brucei CTPS is more pronounced when the enzyme is preincubated with the drug in the presence of nucleotide substrates than in the absence of substrates
adenosine 5'-[beta,gamma-imido]triphosphate
-
poor inhibitor compared to ATPgammaS
alpha-amino-3-chloro-4,5-dihydro-5-isoxazoleacetic acid
-
reduces the parasite CTP level even further and inhibits trypanosome proliferation in vitro and in Trypanosoma brucei-infected mice
Cu2+
-
inhibition is not reversed by EDTA, in presence of dithiothreitol inhibition at concentrations below 0.2 mM
DL-DELTA1-pyrroline 5-carboxylate
-
0.125 mM, complete inhibition of ammonium chloride-dependent CTP synthesis
Gln
-
inhibition of hydroxylamine-dependent N4-OH CTP synthesis in presence of GTP
glutamate gamma-semialdehyde
-
potent linear mixed-type inhibitor, competitive with respect to ammonia, no inhibition of the mutant enzyme C379A
guanosine 5'-tetraphosphate
no guanosine, kact: 4/sec, KA: 0.19 mM, Ki: 0.5 mM; the GTP analogue is capable of inhibiting Gln-dependent CTP formation at over 0.15 mM
ITP
no guanosine, kact: 5.2/sec, KA: 2.9 mM, Ki: 4.5 mM; the GTP analogue is capable of inhibiting Gln-dependent CTP formation at over 0.15 mM
L-2-pyrrolidone 5-carboxylate
-
weak competitive inhibition of the reaction with ammonia as substrate, no significant inhibition with glutamine as substrate
NH4Cl
-
substrate inhibition , a significant part of the inhibition can be shown to be due to the increase in ionic strength with increasing substrate concentrations
Ni2+
-
in presence of dithiothreitol inhibition at concentrations below 0.2 mM
O-methylguanosine 5'-triphosphate
no guanosine, kact: 2.8/sec, KA: 0.13 mM, Ki: 0.29 mM
O6-methyl-GTP
the GTP analogue is capable of inhibiting Gln-dependent CTP formation at over 0.15 mM
pyrrole-2-carboxylate
-
weak competitive inhibition of the reaction with ammonia as substrate, no significant inhibition with glutamine as substrate
S-nitroso-L-cysteine
specific irreversible inhibitor,inhibits the activity by 94%
S-nitroso-L-homocysteine
specific irreversible inhibitor, inhibits the activity by 90%
Zn2+
-
inhibition is reversed by EDTA, in presence of dithiothreitol inhibition at concentrations below 0.2 mM
6-diazo-5-oxo-L-norleucine
-
reduces the parasite CTP level even further and inhibits trypanosome proliferation in vitro and in Trypanosoma brucei-infected mice
CTP
-
the enzyme from T lymphoblast S49 cells is refractory to complete inhibition by CTP
CTP
-
product inhibition of wild-type enzyme, mutant E161K is less sensitive to CTP product inhibition
CTP
-
IC50 for the native enzyme: 0.12 in presence of 0.5 mM ATP, 0.22 mM in presence of 1 mM ATP. IC50 for the phosphorylated enzyme: 0.21 mM in presence of 0.5 mM ATP, 0.31 mM in presence of 1 mM ATP
cyclopentenyl cytosine
-
the inhibitor has a cytostatic effect on lymphoblasts of children with acute lymphocytic leukemia
cyclopentenyl cytosine
-
a highly specific inhibitor of CTPS1, in vivo inhibition leads to alterations in the cell nuclei and microtubule network, overview
GTP
-
inhibition of glutamine-dependent CTP formation above 0.15 mM, inhibition of glutamine-dependent CTP formation in a concentration-dependent manner
GTP
0.1 mM guanosine, kact: 10.3/sec, KA: 0.088 mM, Ki: 0.22 mM; 0.2 mM guanosine, kact: 8.2/sec, KA: 0.078 mM, Ki: 0.12 mM; allosteric effector, structural requirements for activation are stringent, but requirements for inhibition are lax. GTP promotes Gln hydrolysis but inhibits Gln-dependent CTP formation at concentrations of over 0.15 mM; no guanosine, kact: 10.6/sec, KA: 0.081 mM, Ki: 0.28 mM
GTP
-
GTP acts a positive allosteric effector for Gln-dependent CTP formation. However, at concentrations exceeding 0.15 mM, GTP inhibits Gln-dependent CTP formation. Moreover, GTP is an inhibitor of NH3-dependent CTP formation at all concentrations
GTP
-
inhibits glutamine-dependent CTP formation at concentrations above 0.2 mM
additional information
-
enzyme loses activity at ionic strengths higher than 0.4 M
-
additional information
GTP analogues inhibite NH3-and Gln-dependent CTP-formation, often in a cooperative manner, to a similar extent as they activate it with IC50 values of 0.2-0.5 mM, the inhibition appears to be due solely to the purine base, binding structures and kinetics, overview. Inhibitor structure-activity study, overview
-
additional information
-
GTP analogues inhibite NH3-and Gln-dependent CTP-formation, often in a cooperative manner, to a similar extent as they activate it with IC50 values of 0.2-0.5 mM, the inhibition appears to be due solely to the purine base, binding structures and kinetics, overview. Inhibitor structure-activity study, overview
-
additional information
incubation of EcCTPS modified by CysNO and HcyNO with 5 mM DTT for 30 min at 37°C reveals that 88% and 97%, respectively, of the original activity can be recovered; it is shown that in the presence of 1 mM Gln, S-nitroso-L-cysteine reduces the enzymatic activity by 88% and by 32% in the presence of 10 mM Gln. Similar studies with S-nitroso-L-homocysteine result in reduction of the activity by 43% and 19%, respectively. The results suggest that the substrate Gln competitively protects the active site of EcCTPS from the modification with S-nitroso-L-cysteine and S-nitroso-L-homocysteine.; no inhibition by S-nitrosoglutathione presumably due to its inability to enter the actve site of the enzyme
-
additional information
-
inhibition by xanthine and derivatives, no inhibition by allantoin, an intact purine ring with anionic character favors inhibition. In general, methylation of the purine does not significantly affect inhibition
-
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
6-thio-GTP
the GTP analogue is capable of activating Gln-dependent CTP formation
6-thioguanosine 5'-triphosphate
0 mM guanosine, kact: 8.5/sec, KA: 0.035 mM, Ki: 0.27 mM
dithiothreitol
-
required for optimal activity, above 50 mM increase activity to 223% mM
O-methylguanosine 5'-triphosphate
0 mM guanosine, kact: 2.8/sec, KA: 0.13 mM, Ki: 0.29 mM
O6-methyl-GTP
the GTP analogue is capable of activating Gln-dependent CTP formation
2'-deoxy-GTP
the GTP analogue is capable of activating Gln-dependent CTP formation
2-mercaptoethanol
-
required for optimal activity, above 50 mM increase activity to 221%
GTP
-
required as allosteric effector to promote reaction with glutamine as substrate, GTP functions by stabilizing the protein conformation that binds the tetrahedral intermediate(s) formed during glutamine hydrolysis
GTP
allosteric effector, structural requirements for activation are stringent, but requirements for inhibition are lax. GTP promotes Gln hydrolysis but inhibits Gln-dependent CTP formation at concentrations of over 0.15 mM
GTP
-
GTP acts a positive allosteric effector for Gln-dependent CTP formation. However, at concentrations exceeding 0.15 mM, GTP inhibits Gln-dependent CTP formation. Moreover, GTP is an inhibitor of NH3-dependent CTP formation at all concentrations
GTP
-
the binding of GTP to the allosteric site promotes coordination of the phosphorylation of UTP and hydrolysis of glutamine for optimal efficiency in CTP synthesis rather than just acting to increase the rate of glutamine hydrolysis itself
GTP
-
not essential, but acts as activator on the glutamine reaction, optimal activation at 1 mM
GTP
-
activates glutamine-dependent CTP formation at concentrations below 0.2 mM
guanosine 5'-tetraphosphate
0 mM guanosine, kact: 4/sec, KA: 0.19 mM, Ki: 0.5 mM
guanosine 5'-tetraphosphate
the GTP analogue is capable of activating Gln-dependent CTP formation
ITP
the GTP analogue is capable of activating Gln-dependent CTP formation
additional information
activation potency in descending order: GTP = 6-thio-GTP, ITP = guanosine 5'-tetraphosphate, O6-methyl-GTP, 2'-deoxy-GTP, no activation with guanosine, GMP, GDP, 2',3'-dideoxy-GTP, acycloguanosine, and acycloguanosine monophosphate, indicating that the 5'-triphosphate, 2'-OH, and 3'-OH are required for full activation, binding structures and kinetics, overview
-
additional information
-
activation potency in descending order: GTP = 6-thio-GTP, ITP = guanosine 5'-tetraphosphate, O6-methyl-GTP, 2'-deoxy-GTP, no activation with guanosine, GMP, GDP, 2',3'-dideoxy-GTP, acycloguanosine, and acycloguanosine monophosphate, indicating that the 5'-triphosphate, 2'-OH, and 3'-OH are required for full activation, binding structures and kinetics, overview
-
additional information
binding of the substrates ATP and UTP, or the product CTP, promotes oligomerization of CTPS from inactive dimers to active tetramers, Gly142 is critical for nucleotide-dependent oligomerization of CTPS to active tetramers
-
additional information
-
binding of the substrates ATP and UTP, or the product CTP, promotes oligomerization of CTPS from inactive dimers to active tetramers, Gly142 is critical for nucleotide-dependent oligomerization of CTPS to active tetramers
-
additional information
-
the URA7-encoded enzyme is phosphorylated by protein kinases A and C at Ser424, and these phosphorylations stimulate CTP synthetase activity and increase cellular CTP levels and the utilization of the Kennedy pathway
-
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.206
glutamine
wild-type protein, presence of 3 mM ATP, 2 mM UTP
0.345
glutamine
wild-type protein, presence of 1 mM ATP, 1 mM UTP
0.04
UTP
-
pH 8.0, 30°C, phosphorylated enzyme, in presence of 2 mM ATP
0.07
UTP
-
pH 8.0, mutant enzyme S354A, kinetic constant determined with 0.5 mM ATP
0.08
UTP
-
pH 8.0, 30°C, native enzyme or phosphorylated enzyme, in presence of 0.5 mM ATP
0.1
UTP
-
pH 8.0, mutant enzyme S330A, kinetic constant determined with 0.5 mM ATP
additional information
additional information
kinetic mechanism, activation and inhibition kinetics
-
additional information
additional information
-
kinetic mechanism, activation and inhibition kinetics
-
additional information
additional information
kinetics of wild-type and mutants
-
additional information
additional information
-
kinetics of wild-type and mutants
-
additional information
additional information
-
positive cooperativity for ATP and UTP
-
additional information
additional information
-
maximally active at physiological concentrations of ATP, GTP, and glutamine, whereas the Km and IC50 values for the substrate UTP and the product CTP, respectively, are close to their physiological concentrations
-
additional information
additional information
maximally active at physiological concentrations of ATP, GTP, and glutamine, whereas the Km and IC50 values for the substrate UTP and the product CTP, respectively, are close to their physiological concentrations
-
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
8.5
6-thioguanosine 5'-triphosphate
no guanosine, kact: 8.5/sec, KA: 0.035 mM, Ki: 0.27 mM
0.0058
2',3'-dialdehyde adenosine 5'-triphosphate
K306A: replacement of lysine 306 by alanine reduces the rate of 2',3'-dialdehyde adenosine 5'-triphosphate-dependent inactivation
0.054
2',3'-dialdehyde adenosine 5'-triphosphate
in the presence of 10 mM UTP
0.08
UTP
pH 8.0, 37°C, mutant G142A with L-glutamine, in presence of 0.25 mM GTP
0.67
UTP
pH 8.0, 37°C, mutant G143A with L-glutamine, in presence of 0.25 mM GTP
4.2
UTP
pH 8.0, 37°C, mutant G146A with L-glutamine, in presence of 0.25 mM GTP
5
UTP
pH 8.0, 37°C, wild-type enzyme with L-glutamine, in presence of 0.25 mM GTP
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Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.33
1-beta-D-ribofuranosyl-2-thiouracil 5'-triphosphate
-
pH 7.2, inhibition of glutamine reaction
0.08
2-Thiocytidine 5'-triphosphate
-
pH 7.2, inhibition of glutamine reaction and ammonia reaction
0.182
Gln
-
30°C, pH 8.0, inhibition of hydroxylamine-dependent N4-OH CTP synthesis in presence of GTP
3.36
2',3'-dialdehyde adenosine 5'-triphosphate
in the presence of 10 mM UTP
3.7
2',3'-dialdehyde adenosine 5'-triphosphate
K306A: replacement of lysine 306 by alanine reduces the rate of 2',3'-dialdehyde adenosine 5'-triphosphate-dependent inactivation
0.1
2-thiouridine 5'-triphosphate
-
pH 7.2, inhibition of glutamine reaction
0.25
2-thiouridine 5'-triphosphate
-
pH 7.2, inhibition of ammonia reaction
additional information
additional information
no saturation kinetic is obtained with S-nitroso-L-homocysteine
-
additional information
additional information
nucleotide and nucleoside analogues of GTP and guanosine, respectively, all inhibit NH3- and Gln-dependent CTP formation to a similar extent (IC50 : 0.20.5 mM). This inhibition appears to be due solely to the purine base and is relatively insensitive to the identity of the purine with the exception of inosine, ITP, and adenosine (IC50 : 412 mM). 8-Oxoguanosine is the best inhibitor identified (IC50 : 0.08 mM).
-
additional information
additional information
-
nucleotide and nucleoside analogues of GTP and guanosine, respectively, all inhibit NH3- and Gln-dependent CTP formation to a similar extent (IC50 : 0.20.5 mM). This inhibition appears to be due solely to the purine base and is relatively insensitive to the identity of the purine with the exception of inosine, ITP, and adenosine (IC50 : 412 mM). 8-Oxoguanosine is the best inhibitor identified (IC50 : 0.08 mM).
-
additional information
additional information
kinetic mechanism, activation and inhibition kinetics
-
additional information
additional information
-
kinetic mechanism, activation and inhibition kinetics
-
additional information
additional information
-
inhibition kinetics. Multisite inhibition of CTPS-catalyzed NH3-dependent CTP formation by caffeine
-
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IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
additional information
Escherichia coli
-
xanthine and related compounds inhibit CTPS activity with IC50 = 0.16-0.58 mM. The presence of an 8-oxo function enhances the inhibition to IC50 = 0.060-0.121 mM. Raising the pH from 8.0 to 8.5 results in slightly increased inhibition of NH3-dependent CTP formation by the xanthines
-
0.33
2'-deoxy-guanosine
Escherichia coli
IC50 for inhibition of Gln-dependent CTP formation
0.45
2'-deoxy-guanosine
Escherichia coli
IC50 for inhibition of NH3-dependent CTP formation
0.11
2,6-diaminopurine riboside
Escherichia coli
IC50 for inhibition of Gln-dependent CTP formation
0.18
2,6-diaminopurine riboside
Escherichia coli
IC50 for inhibition of NH3-dependent CTP formation
0.22
2-aminopurine riboside
Escherichia coli
IC50 for inhibition of Gln-dependent CTP formation
0.26
2-aminopurine riboside
Escherichia coli
IC50 for inhibition of NH3-dependent CTP formation
0.34
3'-deoxy-guanosine
Escherichia coli
IC50 for inhibition of Gln-dependent CTP formation
0.4
3'-deoxy-guanosine
Escherichia coli
IC50 for inhibition of NH3-dependent CTP formation
0.39
6-thioguanine
Escherichia coli
IC50 for inhibition of Gln-dependent CTP formation
0.61
6-thioguanine
Escherichia coli
IC50 for inhibition of NH3-dependent CTP formation
0.23
6-thioguanosine
Escherichia coli
IC50 for inhibition of Gln-dependent CTP formation
0.4
6-thioguanosine
Escherichia coli
IC50 for inhibition of NH3-dependent CTP formation
0.08
8-oxoguanosine
Escherichia coli
IC50 for inhibition of Gln-dependent CTP formation
0.13
8-oxoguanosine
Escherichia coli
IC50 for inhibition of NH3-dependent CTP formation
0.11
8-oxoguanosine 5'-triphosphate
Escherichia coli
IC50 for inhibition of Gln-dependent CTP formation
0.15
8-oxoguanosine 5'-triphosphate
Escherichia coli
IC50 for inhibition of NH3-dependent CTP formation
0.33
acycloguanosine
Escherichia coli
IC50 for inhibition of Gln-dependent CTP formation
0.45
acycloguanosine
Escherichia coli
8.16 mM NH3, IC50 for inhibition of NH3-dependent CTP formation
0.45
acycloguanosine
Escherichia coli
IC50 for inhibition of NH3-dependent CTP formation
0.47
acycloguanosine
Escherichia coli
5.44 mM NH3, IC50 for inhibition of NH3-dependent CTP formation
0.48
acycloguanosine
Escherichia coli
2.72 mM NH3, IC50 for inhibition of NH3-dependent CTP formation
0.49
acycloguanosine
Escherichia coli
1.36 mM NH3, IC50 for inhibition of NH3-dependent CTP formation
0.31
acycloguanosine monophosphate
Escherichia coli
IC50 for inhibition of Gln-dependent CTP formation
0.41
acycloguanosine monophosphate
Escherichia coli
IC50 for inhibition of NH3-dependent CTP formation
0.21
CTP
Saccharomyces cerevisiae
-
IC50 for the native enzyme: 0.12 in presence of 0.5 mM ATP, 0.22 mM in presence of 1 mM ATP. IC50 for the phosphorylated enzyme: 0.21 mM in presence of 0.5 mM ATP, 0.31 mM in presence of 1 mM ATP
0.29
GDP
Escherichia coli
2.72 mM NH3, IC50 for inhibition of NH3-dependent CTP formation
0.29
GDP
Escherichia coli
5.44 mM NH3, IC50 for inhibition of NH3-dependent CTP formation
0.3
GDP
Escherichia coli
1.36 mM NH3, IC50 for inhibition of NH3-dependent CTP formation
0.23
GMP
Escherichia coli
5.44 mM NH3, IC50 for inhibition of NH3-dependent CTP formation
0.25
GMP
Escherichia coli
2.72 mM NH3, IC50 for inhibition of NH3-dependent CTP formation
0.28
GMP
Escherichia coli
1.36 mM NH3, IC50 for inhibition of NH3-dependent CTP formation
0.29
GMP
Escherichia coli
8.16 mM NH3, IC50 for inhibition of NH3-dependent CTP formation
0.29
GTP
Escherichia coli
5.44 mM NH3, IC50 for inhibition of NH3-dependent CTP formation
0.3
GTP
Escherichia coli
2.72 mM NH3, IC50 for inhibition of NH3-dependent CTP formation
0.3
GTP
Escherichia coli
8.16 mM NH3, IC50 for inhibition of NH3-dependent CTP formation
0.31
GTP
Escherichia coli
1.36 mM NH3, IC50 for inhibition of NH3-dependent CTP formation
0.22
guanosine
Escherichia coli
5.44 mM NH3, IC50 for inhibition of NH3-dependent CTP formation
0.26
guanosine
Escherichia coli
1.36 mM NH3, IC50 for inhibition of NH3-dependent CTP formation
0.26
guanosine
Escherichia coli
2.72 mM NH3, IC50 for inhibition of NH3-dependent CTP formation
0.29
guanosine
Escherichia coli
8.16 mM NH3, IC50 for inhibition of NH3-dependent CTP formation
0.3
guanosine
Escherichia coli
0.30 mM Gln, IC50 for inhibition of Gln-dependent CTP formation
0.32
guanosine
Escherichia coli
10 mM Gln, IC50 for inhibition of Gln-dependent CTP formation
0.32
guanosine
Escherichia coli
2.5 mM Gln, IC50 for inhibition of Gln-dependent CTP formation
0.35
guanosine
Escherichia coli
0.75 mM Gln, IC50 for inhibition of Gln-dependent CTP formation
0.33
guanosine 5'-tetraphosphate
Escherichia coli
IC50 for inhibition of Gln-dependent CTP formation
0.42
guanosine 5'-tetraphosphate
Escherichia coli
IC50 for inhibition of NH3-dependent CTP formation
2.9
ITP
Escherichia coli
8.16 mM NH3, IC50 for inhibition of NH3-dependent CTP formation
3
ITP
Escherichia coli
2.72 mM NH3, IC50 for inhibition of NH3-dependent CTP formation
3.1
ITP
Escherichia coli
5.44 mM NH3, IC50 for inhibition of NH3-dependent CTP formation
3.3
ITP
Escherichia coli
1.36 mM NH3, IC50 for inhibition of NH3-dependent CTP formation
0.17
N-methylguanosine
Escherichia coli
IC50 for inhibition of Gln-dependent CTP formation
0.23
N-methylguanosine
Escherichia coli
IC50 for inhibition of NH3-dependent CTP formation
0.15
O-methylguanosine
Escherichia coli
0.30 mM Gln, IC50 for inhibition of Gln-dependent CTP formation
0.16
O-methylguanosine
Escherichia coli
10 mM Gln, IC50 for inhibition of Gln-dependent CTP formation
0.16
O-methylguanosine
Escherichia coli
2.5 mM Gln, IC50 for inhibition of Gln-dependent CTP formation
0.16
O-methylguanosine
Escherichia coli
IC50 for inhibition of Gln-dependent CTP formation
0.17
O-methylguanosine
Escherichia coli
0.75 mM Gln, IC50 for inhibition of Gln-dependent CTP formation
0.25
O-methylguanosine
Escherichia coli
IC50 for inhibition of NH3-dependent CTP formation
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0.0025
-
specific activity of Escherichia coli expressed enzyme, after a 20 min incubation with protein kinase C, the activity of the CTP synthetase was stimulated 95fold
additional information
additional information
-
fast assay allows the processing of a large number of samples
additional information
a structure-activity study using a variety of GTP and guanosine analogues reveals that only a few GTP analogues are capable of activating Gln-dependent CTP formation to varying degrees: GTP > 6-thio-GTP > ITP> guanosine 5'-tetraphosphate > O-methyl-GTP > 2'-deoxy-GTP. No activation is observed with guanosine, GMP, GDP, 2',3'-dideoxy-GTP, acycloguanosine, and acycloguanosine monophosphate indicating that the 5'-triphosphate, 2'-OH, and 3'-OH are required for full activation. The 2-NH2 group is important in binding recognition while substituents at the 6-position are important in activation.
additional information
-
a structure-activity study using a variety of GTP and guanosine analogues reveals that only a few GTP analogues are capable of activating Gln-dependent CTP formation to varying degrees: GTP > 6-thio-GTP > ITP> guanosine 5'-tetraphosphate > O-methyl-GTP > 2'-deoxy-GTP. No activation is observed with guanosine, GMP, GDP, 2',3'-dideoxy-GTP, acycloguanosine, and acycloguanosine monophosphate indicating that the 5'-triphosphate, 2'-OH, and 3'-OH are required for full activation. The 2-NH2 group is important in binding recognition while substituents at the 6-position are important in activation.
additional information
-
CTP synthetase 1 activity of the T455A mutant enzyme is 2fold higher than the wild type enzyme. T455A mutation causes a 44% decrease in the amount of human CTP synthetase 1 that is phosphorylated in Saccharomyces cerevisiae cells, accompanied by a 2.5fold increase in the cellular concentration of CTP and a 1.5-fold increase in the choline-dependent synthesis of phosphatidylcholine
additional information
low serum is found to decrease CTPS1 activity, and incubation with the glycogen synthase kinase 3 inhibitor indirubin-3-monoxime protects against this decrease in activity. Incubation with an alkaline phosphatase increases CTPS1 activity in a time-dependent manner, demonstrating that phosphorylation inhibits CTPS1 activity
additional information
-
low serum is found to decrease CTPS1 activity, and incubation with the glycogen synthase kinase 3 inhibitor indirubin-3-monoxime protects against this decrease in activity. Incubation with an alkaline phosphatase increases CTPS1 activity in a time-dependent manner, demonstrating that phosphorylation inhibits CTPS1 activity
additional information
-
S462A mutation results in 61%-reduced CTP synthetase 1 phosphorylation. The Saccharomyces cerevisiae-expressed and purified S462A mutant enzyme exhibits a 2fold reduction in CTP synthetase 1 activity, whereas the purified T455A mutant enzyme exhibited a 2fold elevation in CTP synthetase 1 activity, implying that that protein kinase C phosphorylation at Ser462 stimulates human CTP synthetase 1 activity, whereas phosphorylation at Thr455 inhibits activity
additional information
-
T455A mutation results in 58%-reduced CTP synthetase 1 phosphorylation. The Saccharomyces cerevisiae-expressed and purified S462A mutant enzyme exhibits a 2fold reduction in CTP synthetase 1 activity, whereas the purified T455A mutant enzyme exhibited a 2fold elevation in CTP synthetase 1 activity, implying that that protein kinase C phosphorylation at Ser462 stimulates human CTP synthetase 1 activity, whereas phosphorylation at Thr455 inhibits activity
additional information
-
Thr 455 is identified as a major site of phosphorylation by protein kinase A, phosphorylation at Thr455 results in the inhibition of activity in vitro and in vivo. Data indicate that phosphorylation at Thr455 attenuates the choline-dependent synthesis of phosphatidylcholine when CTP synthetase 1 enzyme is expressed in Saccharomyces cerevisiae
additional information
-
during the purification, the specific activity of the enzyme preparation increased 836fold and the total yield is 28%
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
10.3 - 10.4
-
ammonia-dependent activity,maximal activity with NH4+ is at least 20% greater than with glutamine
8.7
-
reaction with NH4+, glutamine-dependent reaction activated by Mg2+ or Mn2+
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pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
7.3 - 9.5
-
pH 7.3: about 35% of maximal activity, pH 9.5: about 40% of maximal activity
7.5 - 9
-
pH 7.5: about 70% of maximal activity, pH 9.0: about 55% of maximal activity, dUTP-dependent activity
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TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
37
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TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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pI VALUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
brenda
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SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
-
significantly higher activity is detected in leukaemic cells of children suffering from acute lymphocytic leukemia than in lymphocytes of healthy controls
brenda
Myc protein levels correlate with cytoophidium abundance in follicle epithelia
brenda
-
highest activity is found in thombocytes, followed by monocytes, lymphocytes, granulocytes and erythrocytes
brenda
additional information
-
in the liver tumours of slow and medium growth rate the specific activity of CTP synthetase is increased 1.8fold to 3.6fold that of normal rat livers, the activity increases from 4.6fold to 11.2fold in rapidly growing hepatomas
brenda
cytoophidia are present in all major cell types in the ovary
brenda
additional information
-
transcribed and translated primarily during the mid and late stages of the Chlamydial growth cycle
brenda
additional information
-
pfCTP synthetase exists at all stages of intracellular parasite life cycle
brenda
additional information
-
the URA7 mRNA is 2-fold more abundant than the URA8 transcript
brenda
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LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
additional information
-
filamentary structure present in all major cell types, exhibits a characteristically elongated and serpentine form
brenda
-
CTP synthase isoform C forms the cytoophidium
brenda
in cytoophidium assembly, formation of heteromeric CTP synthase filaments takes place, which is disrupted by CTP synthase carrying a mutated N-terminal alanine residue
brenda
-
CTP synthase isoform B distributes diffusely in the cytoplasm
brenda
-
CTP synthase isoform A localizes in the nucleus
brenda
additional information
-
CTP synthase forms filaments in Caulobacter crescentus, and the filaments it forms regulate the curvature of Caulobacter crescentus cells independently of its catalytic function. The morphogenic role of CTP synthase requires its functional interaction with the intermediate filament, crescentin
-
brenda
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GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
malfunction
-
mutations disrupting CTP synthase isoform C or isoform A expression results in cytoophidium disassembly
physiological function
physiological function
-
central role of CTP in the biosynthesis of nucleic acids, phospholipids, and sialic acid
physiological function
-
CTP synthase forms filaments in Caulobacter crescentus, and the filaments it forms regulate the curvature of Caulobacter crescentus cells independently of its catalytic function. The morphogenic role of CTP synthase requires its functional interaction with the intermediate filament, crescentin
physiological function
-
CTP synthase protein molecules form filamentous structures termed cytoophidia or CTP synthase filaments in the cytoplasm and nucleus
physiological function
-
CTP synthase protein molecules form filamentous structures termed cytoophidia or CTP synthase filaments in the cytoplasm and nucleus
physiological function
destabilization of the active tetrameric form of the enzyme increases filament formation. The sites responsible for feedback inhibition and allosteric activation control filament length, implying that multiple regions of the enzyme can influence filament structure. Blocking catalysis without disrupting the regulatory sites of the enzyme does not affect filament formation or length
physiological function
in cytoophidium assembly, formation of heteromeric CTP synthase filaments takes place, which is disrupted by CTP synthase carrying a mutated N-terminal alanine residue
physiological function
in ovarian germline cells CTPS filaments are catalytically active and their assembly is regulated by the non-receptor tyrosine kinase DAck. Egg chambers from flies deficient in DAck catalytic activity exhibit disrupted CTPS filament architecture and morphological defects that correlate with reduced fertility. Ovaries from these flies exhibit reduced levels of total RNA
physiological function
Myc protein levels correlate with cytoophidium abundance in follicle epithelia. Reducing Myc levels results in cytoophidium loss and small nuclear size in follicle cells, while overexpression of Myc increases the length of cytoophidia and the nuclear size of follicle cells. Ectopic expression of Myc induces cytoophidium formation in late stage follicle cells. Furthermore, knock-down of CTPsyn is sufficient to suppress the overgrowth phenotype induced by Myc overexpression
physiological function
-
overexpression of CTPS by in utero electroporation in the embryonic mouse brain induces formation of cytoophidia in developing cortical neurons and impairs neuronal migration. The increase of cytoophidia accelerates neuronal differentiation and inhibits neural progenitor cell proliferation by reducing their mitotic activity. The cytoophidia diffuse during the early G1-phase of the cell cycle
physiological function
-
destabilization of the active tetrameric form of the enzyme increases filament formation. The sites responsible for feedback inhibition and allosteric activation control filament length, implying that multiple regions of the enzyme can influence filament structure. Blocking catalysis without disrupting the regulatory sites of the enzyme does not affect filament formation or length
-
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PDB
SCOP
CATH
UNIPROT
ORGANISM
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MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
210000
52000
-
4 * 52000, enzyme exists as tetramer in presence of UTP, Mg2+ and ATP
70000
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SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
dimer
tetramer
additional information
dimer
-
the enzyme is a dimer of 108000 Da, the dimer associates to form a tetramer in the presence of either ATP or UTP
dimer
Escherichia coli B / ATCC 11303
-
the enzyme is a dimer of 108000 Da, the dimer associates to form a tetramer in the presence of either ATP or UTP
-
dimer
crystallization data and sedimentation-ultracentrifugation experiments
tetramer
-
4 * 52000, enzyme exists as tetramer in presence of UTP, Mg2+ and ATP
tetramer
-
UTP and ATP are responsible for the tetramerization and activation of the inactive dimeric form of the enzyme. UTP is absolutely required for the tatramerization of the enzyme when ATP is present at a saturating concentration
tetramer
-
the enzyme exists as an inactive dimer in the absence of ATP and UTP. In the presence of saturating concentrations of ATP and UTP, the CTP synthetase protein exists as an active tetramer. Increasing concentrations of ATP and UTP cause a dose-dependent conversion of the dimeric species to a tetramer. Tetramerization is dependent on UTP and Mg2+ ions. ATP facilitates the UTP-dependent teramerization of CTP synthetase by a mechanism that involves the ATP-dependent phosphorylation of UTP catalyzed by the enzyme
additional information
-
the enzyme can dissociate to an apparently inactive monomer. The dissociation is reversible and the rate of association is slow
additional information
binding of the substrates ATP and UTP, or the product CTP, promotes oligomerization of CTPS from inactive dimers to active tetramers, Gly142 is critical for nucleotide-dependent oligomerization of CTPS to active tetramers. Oligomerization of Gly mutant enzymes, overview
additional information
-
binding of the substrates ATP and UTP, or the product CTP, promotes oligomerization of CTPS from inactive dimers to active tetramers, Gly142 is critical for nucleotide-dependent oligomerization of CTPS to active tetramers. Oligomerization of Gly mutant enzymes, overview
additional information
-
CTP synthetase oligomerizes to a tetramer in the presence of its substrates UTP and ATP
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POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
phosphoprotein
ubiquitination
reversible ubiquitination regulates the dynamic assembly of the filamentous structures of Drosophila CTPsyn. The proto-oncogene Cbl, an E3 ubiquitin ligase, controls CTPsyn filament formation in endocycles. The E3 ligase activity of Cbl is required for CTPsyn filament formation, and Cbl does not affect the protein levels of CTPsyn. In the absence of Cbl or with knockdown of CTPsyn, the progression of the endocycle-associated S phase is impaired. Overexpression of wild-type, but not enzymatically inactive CTPsyn, rescues the endocycle defect in Cbl mutant cells
phosphoprotein
-
CTP synthetase is phosphorylated by protein kinase C which leads to 95fold higher activity of CTP synthetase 1. Phosphopeptide mapping and phosphoamino acid analyses shows that CTP synthetase 1 is phosphorylated on multiple serine and threonine residues. It is indicated that protein kinase C phosphorylation at Ser462 stimulates human CTP synthetase 1 activity, whereas phosphorylation at Thr455 inhibits activity
phosphoprotein
it is shown that Ser-574 and Ser-575 are phosphorylated residues. Mutation of Ser-571 demonstrates that Ser-571 is the major site phosphorylated by glycogen synthase kinase 3 in intact human embryonic kidney 293 cells by glycogen synthase kinase 3 in vitro. Mutation of Ser-575 prevents phosphorylation of Ser-571, suggesting phosphorylation of Ser-575 is necessary for priming the glycogen synthase kinase 3 phosphorylation of Ser-571
phosphoprotein
stimulation/inhibition of protein kinase A and protein kinase C does not alter the phosphorylation of endogenous human CTPS1 in human embryonic kidney 293 cells. Low serum conditions increased phosphorylation of endogenous CTPS1 and phosphorylation is inhibited by the glycogen synthase kinase 3 (GSK3) inhibitor indirubin-3-monoxime and GSK3beta short interfering RNAs, demonstrating the involvement of glycogen synthase kinase 3 in phosphorylation of endogenous human CTPS1
phosphoprotein
-
the CTPS1-encoded enzyme is phosphorylated by protein kinases A and C at Thr455, and these phosphorylations stimulate CTP synthetase activity and increase cellular CTP levels and the utilization of the Kennedy pathway
phosphoprotein
Low serum treatment increases CTPS2 phosphorylation. Ser568 and Ser571 are two major phosphorylation sites, and Ser568 is phosphorylated by casein kinase 1 both in vitro and in vivo. Mutation of S568A but not S571A significantly increases CTPS2 activity. The S568A mutation has a greater effect on the glutamine than ammonia-dependent activity
phosphoprotein
-
phosphorylation of CTP synthetase by protein kinase A results in the stimulation of CTP synthetase activity. The mechanism of stimulation involves an increase in Vmax of the reaction and an increase of the enzyme affinity for ATP
phosphoprotein
-
phosphorylation of the purified native CTP synthetase with protein kinase A and protein kinase C facilitates the nucleotide-dependent tetramerization. Dephosphorylation of native CTP-desynthetase with alkaline phosphatase prevents the nucleotide-dependent tetramerization of the enzyme
phosphoprotein
-
the enzyme is phosphorylated and stimulated by protein kinase C. Phosphorylation of CTP synthetase on Ser36, Ser330, Ser354, and Ser454 regulates the levels of CTP and phosphatidylcholine synthesis
phosphoprotein
-
the URA7-encoded enzyme is phosphorylated at Ser424 by protein kinases A and C, and these phosphorylations stimulate CTP synthetase activity and increase cellular CTP levels and the utilization of the Kennedy pathway
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CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
2.3 A resolution crystal structure of apoenzyme using Hg-multiwavelength anomalous dispersion phasing, vapor diffusion method, crystals belong to space group P2(1)2(1)2(1), in which each bifunctional monomer contains a dethiobiotin synthetase-like amidoligase N-terminal domain and a type 1 glutamine amidotransferase C-terminal domain
crystal structure analysis discloses a homotetrameric structure and each active site is formed by three different subunits. Sulfate ions bound to the active sites indicate the positions of phosphate-binding sites for the substrates ATP and UTP and the feedback inhibitor CTP. Together with earlier structures of bacterial CTPS, the human CTPS structure provides an extended understanding of the structurefunction relationship of CTPS-family members
-
the protein is crystallized by the hanging-drop method, equilibrating 1 ml protein solution and 1 ml of a solution containing 0.1 M Tris pH 8.8, 1.2 M (NH4)2SO4 and 50 mM malonic acid against 0.5 ml of a well solution containing 0.1 M Tris pH 8.8 and 1.2 M (NH4)2SO4. Crystals with a maximum dimension of 0.03 nm formed in 3 d
-
dimeric form of CTP synthase at 2.5 A resolution. A comparison of the dimeric interface with the intermolecular interfaces in the tetrameric structures of Thermus thermophilus CTP synthase and Escherichia coli CTP synthase shows that the dimeric interfaces are almost identical in the three systems. Residues that are involved in the tetramerization of Sulfolobus solfataricus CTP synthase all have large thermal parameters in the dimeric form and undergo substantial movement upon tetramerization
crystal structures of Thermus thermophilus HB8 CTP synthetase as a tetramer in its native form, CTP synthetase as complex with 3 SO42- and CTP synthetase as a complex with glutamine. Crystallization at 20 °C using vapor-diffusion method. The space group is I222 with cell dimensions of a = 90.4 A, b = 117.1 A and c = 142.3 A
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PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
D149E
DELTA20
deletion of the N-terminal 20 amino acids alone is sufficient to disrupt cytoophidium assembly, mutant protein forms distinct cytoplasmic structures which appear as large clusters
C379A
-
mutant enzyme is fully active with ammonia but has no glutamine-dependent activity, no inhibition by glutamate gamma-semialdehyde
D107A
-
enzyme exhibits wild-type NH3-dependent activity and affinity for glutamine, but impaired glutamine-dependent CTP formation, affinity of the mutant enzyme for GTP is reduced 2-4fold
E103A
-
mutant enzyme exhibits no glutamine-dependent activity and is only partially active with NH3
G110A
-
affinity of the mutant enzyme for GTP is reduced 2-4fold, enzyme exhibits wild-type NH3-dependent activity and affinity for glutamine, but impaired glutamine-dependent CTP formation
G142A
site-directed mutagenesis, inactive mutant with both ammonia and glutamine
G143A
site-directed mutagenesis, kcat/Km for ammonia-dependent and glutamine-dependent CTP formation by mutant G143A are reduced by 22fold and 16fold, respectively, compared to the wild-type enzyme. The mutant is able to form active tetramers in the presence of ATP and UTP
G146A
site-directed mutagenesis, kcat/Km for ammonia-dependent and glutamine-dependent CTP formation by mutant G143A are reduced by 1.4fold and 1.8fold, respectively, compared to the wild-type enzyme. The mutant is able to form active tetramers in the presence of ATP and UTP
G351A
-
mutation increases lability of the enzyme, mutant enzyme is not overproduced because of apparent instability and proteolytic degradation
G352C
-
mutation increases lability of the enzyme, mutation abolishes the capacity to form the covalent glutaminyl-cysteine379 catalytic intermediate, thus preventing glutamine amide transfer function, mutant enzyme is not overproduced because of apparent instability and proteolytic degradation
G352P
-
mutation increases lability of the enzyme, mutation abolishes the capacity to form the covalent glutaminyl-cysteine379 catalytic intermediate, thus preventing glutamine amide transfer function, mutant enzyme is not overproduced because of apparent instability and proteolytic degradation
H118A
-
mutant enzyme exhibits no glutamine-dependent activity and is only partially active with NH3
K102A
-
mutant enzyme exhibits wild-type activity with respect to NH3 and glutamine
K297A
replacement of lysine 297 by alanine does not affect NH3-dependent CTP formation, relative to wild-type CTPS, but reduces kcat for the glutaminase activity 78fold
K306A
replacement of lysine 306 by alanine reduces the rate of 2',3'-dialdehyde adenosine 5'-triphosphate-dependent inactivation (Kinact = 0.0058/sec, Ki = 3.7 mM) and reduces the apparent affinity for CTPS for both ATP and UTP by 2fold. The efficiency of K306A-catalyzed glutamine-dependent CTP formation is also reduced 2fold while near wild type activity is observed when NH3 is the substrate. These findings suggest that Lys 206 is not essential for ATP binding, but does play a role in bringing about the conformational changes that mediate interactions between ATP and UTP sites, and between the ATP-binding site and the glutamine amide transfer domain
L109A
R104A
-
mutant enzyme exhibits no glutamine-dependent activity and is only partially active with NH3
R105A
-
enzyme exhibits wild-type NH3-dependent activity and affinity for glutamine, but impaired glutamine-dependent CTP formation
E579A
HEK 293 cells: no effect on phosphorylation of CTPS1 by glycogen synthase kinase 3
S462A/T455A
-
S462A and T455A mutations result in a decreased CTP synthetase 1 phosphorylation which appear to be much less than of the individual mutant enzymes S462A or T455A
S568A
2fold increase in Km value for UTP. Mutation of S568A significantly increases CTPS2 activity. The S568A mutation has a greater effect on the glutamine than ammonia-dependent activity
S571I
Ser-571 is the major site phosphorylated by glycogen synthase kinase 3 in intact human embryonic kidney 293 cells
S571I/S574A
phosphorylation by glycogen synthase kinase 3 does not show altered incorporation of phosphate compared with S571I alone
S571I/S574A/S575A
phosphorylation by glycogen synthase kinase 3 shows a slight decrease in the amount of phosphate incorporated into CTPS1 compared with S571I/S575A, suggesting that Ser-574 may serve as a minor secondary site for glycogen synthase kinase 3 phosphorylation
S571I/S575A
phosphorylation by glycogen synthase kinase 3 shows slightly elevated amounts of phosphate incorporated into CTPS1 compared with S571I, suggesting that without the ability to phosphorylate Ser-571 or Ser-575 in vitro, glycogen synthase kinase 3 may phosphorylate an alternative site, albeit to a much lesser extent
S574A/S575A
greatly reduces phosphorylation by glycogen synthase kinase 3
S575A
T455A
-
T455A mutation causes a 78% decrease in protein kinase A phosphorylation
E362Q
-
turnover number for NH4Cl-dependent GTP synthesis reaction is 1.6fold higher than wild-type value
G360A
-
5fold increase in GTP-dependent activation of uncoupled glutamine hydrolysis compared to wild-type enzyme. Turnover number for NH4Cl-dependent GTP synthesis reaction is 1.2fold higher than wild-type value
G360P
-
mutant enzyme shows no GTP activation of the uncoupled glutaminase reaction, about 4fold lower turnover number for NH4Cl-dependent CTP synthesis reaction than wild-type enzyme
R359M
-
mutant enzyme shows no GTP activation of the uncoupled glutaminase reaction. Turnover number for NH4Cl-dependent GTP synthesis reaction is 1.1fold higher than wild-type value
R359P
-
mutant enzyme shows no GTP activation of the uncoupled glutaminase reaction. Turnover number for NH4Cl-dependent GTP synthesis reaction is 1.2fold lower than wild-type value
D70A
mutation in the ATP binding site, mutant exhibits approximately twofold increase in the number of cells containing filaments
E146A
mutation in the ATP binding site, mutant exhibits approximately twofold increase in the number of cells containing filaments
E161K
R381M
mutation in L11 lid, 3.1fold increase in the number of cells forming filaments
R381P
mutation in L11 lid, 3.1fold increase in the number of cells forming filaments
S330A
-
CTP synthetase activity in cells bearing the mutant enzyme is elevated, mutation causes an elevation in the Vmax of the reaction. Mutation does not have a major effect on the oligomerization of CTP synthetase
S354A
-
CTP synthetase activity in extracts from cells bearing the mutant enzyme is reduced when compared with cells bearing the wild-type enzyme, decrease in Vmax of the reaction. The amount of inactive dimeric enzyme form is 98% greater compared to wild-type enzyme
S36A
S454A
-
CTP synthetase activity in extracts from cells bearing the mutant enzyme is reduced when compared with cells bearing the wild-type enzyme. Mutation does not have a major effect on the oligomerization of CTP synthetase
D70A
-
mutation in the ATP binding site, mutant exhibits approximately twofold increase in the number of cells containing filaments
-
E146A
-
mutation in the ATP binding site, mutant exhibits approximately twofold increase in the number of cells containing filaments
-
E161K
-
mutation in CTP binding site, filament formation is completely disrupted and the enzyme can only form foci. Mutation decreases the affinity of the enzyme for CTP
-
S36A
-
mutation in phosphorylation site, causes an approximately twofold decrease in the frequency of filament formation
-
additional information
D149E
-
the single point mutation in the resistant strain L2/CPEC results in loss of CTP feedback inhibition, cells are resistant to the cytotoxic effects of cyclopentenyl cytosine
D149E
-
the single point mutation in the resistant strain L2/CPEC results in loss of CTP feedback inhibition, cells are resistant to the cytotoxic effects of cyclopentenyl cytosine
-
L109A
-
enzyme exhibits wild-type NH3-dependent activity and affinity for glutamine, but impaired glutamine-dependent CTP formation, affinity of the mutant enzyme for GTP is reduced 2-4fold
L109A
uncoupling of the hydrolysis of gamma-glutamyl hydroxamate and nascent NH2OH production from N4-hydroxy-CTP formation is more pronounced with mutant than with wild-type enzyme
S575A
greatly reduces phosphorylation by glycogen synthase kinase 3. Mutation of Ser-575 prevents the phosphorylation of Ser-571, suggesting that phosphorylation of Ser-575 is necessary for priming the glycogen synthase kinase 3 phosphorylation of Ser-571
S575A
-
site-directed mutagenesis the mutant does not interact with peptidyl prolyl isomerase Pin1
E161K
-
specific activity of the mutant URA7-encoded and URA8-encoded enzymes are 2fold greater when compared with the wild-type enzymes. The mutant enzymes are less sensitive to CTP product inhibition with inhibitor constants for CTP of 8.4fold- and 5.5fold greater, respectively, than those of their wild-type counterparts. Cells expressing the E161K mutant enzymes on a multicopy plasmid exhibit an increase in resistance to the pyrimidine poison and cancer therapeutic drug cyclopentenylcytosine and accumulate elevated levels of CTP when compared with cells expressing the wild-type enzymes. Cells expressing the E161K mutation in the URA7-encoded CTP synthetase exhibit an 1.5fold increase in the utilization of the Kennedy pathway for phosphatidylcholine synthesis when compared with control cells. Cells bearing the mutation also exhibit an 1.5fold increase in the synthesis of phosphatidylcholine, 1.3fold for phosphatidylethanolamine and 2fold for phosphatidate and a 1.7fold decrease in synthesis of phosphatidylserine. Cells bearing the e161K mutation exhibit an 1.6fold increase in the ratio of total neutral lipids to phospholipids, an 1.4fold increase in triacylglycerol, a 1.7fold increase in free fatty acids, an1.8fold increase in ergosterol ester and a 1.3fold decrease in diacylgylcerol when compared with control cells
E161K
mutation in CTP binding site, filament formation is completely disrupted and the enzyme can only form foci. Mutation decreases the affinity of the enzyme for CTP
S36A
-
CTP synthetase activity in extracts from cells bearing the mutant enzyme is reduced when compared with cells bearing the wild-type enzyme, decrease in Vmax of the reaction. The amount of inactive dimeric enzyme form is 54% greater compared to wild-type enzyme
S36A
mutation in phosphorylation site, causes an approximately twofold decrease in the frequency of filament formation
additional information
in cytoophidium assembly, formation of heteromeric CTP synthase filaments takes place, which is disrupted by CTP synthase carrying a mutated N-terminal alanine residue. N-terminal swapping, i.e. exchange of the 20 N-terminal amino acids to the residues of human and yeast CTP synthase does not affect CTPsyn cytoophidium assembly
additional information
it can be suggested that the conformational change associated with binding ATP may be transmitted through the L10-alpha11 structural unit (residues 297-312) and thereby mediate effects on the glutaminase activity of CTPS
additional information
-
it can be suggested that the conformational change associated with binding ATP may be transmitted through the L10-alpha11 structural unit (residues 297-312) and thereby mediate effects on the glutaminase activity of CTPS
additional information
-
Escherichia coli CtpS can replace the enzymatic and morphogenic functions of Caulobacter crescentus CtpS
additional information
-
deletion of the C-terminal regulatory domain, residues Ser562-Asp591, of CTPS1 greatly increases the Vmax of the enzyme
additional information
deletion of the C-terminal regulatory domain, residues Ser562-Asp591, of CTPS1 greatly increases the Vmax of the enzyme
additional information
-
the ura7D/ura8D double mutant, that lacks CTP synthetase activity, shows a lethal phenotype, which can be rescued by functional expression of human CTPS1 and CTPS2 genes that encode CTP synthetase enzymes. In an ura8 mutant, CTP levels are 22% lower than in wild-type, whereas the CTP concentration in an ura7 mutant is 64% lower than in wild-type
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pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, 30% glycerol, 100 mM 2-mercaptoethanol, 40% loss of activity after 4 weeks
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-80°C, 30% glycerol, 100 mM 2-mercaptoethanol, 65% of the original activity is retained after 4 weeks
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4°C, 30% glycerol, 100 mM 2-mercaptoethanol, 65% loss of activity after 4 weeks
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4°C, enzyme concentration 1-4 mg/ml, 20 mM sodium phosphate buffer, pH 7.2, 2 mM L-glutamine, 1 mM EDTA, 70 mM 2-mercaptoethanol, 20% glycerol, stable for 3 months
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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
by using talon metal affinity resin, a glass Econo-Column and a Sephadex G-50 column. Proteolysis is performed at 23°C for 40 min. The protein solution is then loaded again on a Talon resin column but this time the flow-through, containing CTPS without His tag, is collected. Finally the protein is loaded onto a 0.2-ml SourceQ15 medium column.
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enzyme is purified by gel filtration on a HiLoad 26/60 Superdex 200 column. Active fractions are pooled and applied to a Mono Q HR 5/5 column. Final purifications step is performed on a HiLoad 26/60 Superdex 75 column. The isolated EcCTPS is found >90% pure as judged by SDS-PAGE and ESI-MS.
His6-tagged human CTP synthetase 1 is purified by loading onto a Ni2+-NTA column
-
His6-tagged wild type and mutant human CTP synthetase 1 enzymes are expressed and purified from yeast cell extracts with Ni2+-NTA resin
-
His6-tagged wild type human CTP synthetase 1 is expressed and purified from Escherichia coli by Ni2+-NTA chromatography and with PorosHQ-ion-exchange chromatography
-
Ni2+-IDA resin column chromatography
the soluble histidine-tagged CTPS is purified using metal ion affinity chromatography and the histidine tag is subsequently removed using thrombin-catalyzed cleavage
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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
a carboxyl-terminal FLAG sequence is introduced after insertion of human CTPS1 into pCDNA 4 myc/his, expression by transient transfection of human embryonic kidney (HEK) 293 cells
cDNA encoding the synthetase domain of human CTPS (isoform 1) is subcloned into the pNIC-Bsa4 vector, the resulting construct codes for a protein with an N-terminal hexahistidine tag with an integrated TEV protease-cleavage site, the entire plasmid is transformed in Escherichia coli BL-21
-
cloned in Escherichia coli BL-21 as an N-terminal 6xHis-tag fusion protein
expressed in Escherichia coli BL21(DE3) cells
expression in a ura7ura8 double mutant that lacks CTP synthetase activity
-
expression in Escherichia coli
expression of the human type II CTP synthetase gene in the CTP synthetase-deficient mutant Escherichia coli JF618 completely abolishes the need for exogenous cytidine for their survival
-
expression of wild-type and mutant enzymes in HEK-293 cells, co-expression with GST-tagged Xenopus laevis peptidyl prolyl isomerase Pin1
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functional expression of human CTPS1 and CTPS2 genes that encode CTP synthetase enzymes in enzyme-deficient Saccharomyces cerevisiae ura7D/ura8D double mutant, complementation
-
genes ura7D and ura8D, DNA and amino acid sequence determination and analysis, expression analysis
-
human CTP synthetase 1 is expressed and purified from a Saccharomyces cerevisiae ura7delta ura8delta double mutant that lacks CTP synthetase activity
-
protein is expressed as a His6-tagged wild type human CTP synthetase 1 fusion protein in Escherichia coli
-
protein is expressed as a His6-tagged wild type human CTP synthetase 1 fusion protein in Saccharomyces cerevisiae
-
synthetase-His6-TEV protease site-tagged Trypanosoma brucei CTP synthetase is expressed in BL21(DE3) pLysS bacteria
-
wild-type and L109A recombinant enzyme are expressed in Escherichia coli BL21(DE3)
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
drug development
medicine
drug development
-
daily injection of acivicin in trypanosome-infected mice suppresses the infection up to one month without any significant loss of weight. Experiments with cultured bloodstream Trypanosoma brucei shows that acivicin is trypanocidal if present at 0.001 nM concentration for at least 4 d. Acivicin may qualify as a drug with desirable properties, i.e. cure within 7 d, according to the current Target Product Profiles of WHO and DNDi
drug development
modifying 2-aminopurine or 2-aminopurine riboside may serve as an effective strategy for developing CTPS inhibitors
drug development
-
structure serves as a basis for structure-based design of anti-proliferative inhibitors
drug development
-
CTP is a recognized target for the development of anticancer, antiviral, and antiprotozoal agents
medicine
-
the enzyme is an target for treatment of African sleeping sickness because the trypanosomes, unlike mammalian cells, cannot compensate for the inhibition of CTP synthetase by the salvage of cytidine. The CTP synthetase inhibitors 6-diazo-5-oxo-L-norleucine and alpha-amino-3-chloro-4,5-dihydro-5-isoxazoleacetic acid reduce the parasite CTP level even further and inhibit trypanosome proliferation in vitro and in Trypanosoma brucei-infected mice
medicine
-
glutamine deficiency induces CTPS cytoophidia in human Huh-7 cells. CTPS cytoophidia are found in various human cancers and some noncancerous tissues. Among 203 tissue samples of hepatocellular carcinoma, 28% exhibited many cytoophidia, whereas no cytoophidia are detected in adjacent noncancerous hepatocytes for all samples
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REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Iyengar, A.; Bearne, S.L.
Aspartate-107 and leucine-109 facilitate efficient coupling of glutamine hydrolysis to CTP synthesis by Escherichia coli CTP synthase
Biochem. J.
369
497-507
2003
Escherichia coli
brenda
Bearne, S.L.; Hekmat, O.; Macdonnell, J.E.
Inhibition of Escherichia coli CTP synthase by glutamate gamma-semialdehyde and the role of the allosteric effector GTP in glutamine hydrolysis
Biochem. J.
356
223-232
2001
Escherichia coli
brenda
Willemoes, M.; Sigurskjold, B.W.
Steady-state kinetics of the glutaminase reaction of CTP synthase from Lactococcus lactis. The role of the allosteric activator GTP incoupling between glutamine hydrolysis and CTP synthesis
Eur. J. Biochem.
269
4772-4779
2002
Lactococcus lactis
brenda
Verschuur, A.C.; van Gennip, A.H.; Muller, E.J.; Voute, P.A.; Vreken, P.; Van Kuilenburg, A.B.
Cytidine triphosphate synthase activity and mRNA expression in normal human blood cells
Biol. Chem.
380
41-46
1999
Homo sapiens
brenda
van Kuilenburg, A.B.; Meinsma, R.; Vreken, P.; Waterham, H.R.; van Gennip, A.H.
Identification of a cDNA encoding an isoform of human CTP synthetase
Biochim. Biophys. Acta
1492
548-552
2000
Escherichia coli (P0A7E5), Escherichia coli, Homo sapiens (P17812), Homo sapiens (Q9NRF8), Homo sapiens, Saccharomyces cerevisiae (P28274), Saccharomyces cerevisiae (P38627), Mus musculus (P70303), Mus musculus (P70698)
brenda
Verschuur, A.C.; Van Gennip, A.H.; Leen, R.; Meinsma, R.; Voute, P.A.; van Kuilenburg, A.B.
In vitro inhibition of cytidine triphosphate synthetase activity by cyclopentenyl cytosine in paediatric acute lymphocytic leukaemia
Br. J. Haematol.
110
161-169
2000
Homo sapiens
brenda
Willemoes, M.; Larsen, S.
Substrate inhibition of Lactococcus lactis cytidine 5'-triphosphate synthase by ammonium chloride is enhanced by salt-dependent tetramer dissociation
Arch. Biochem. Biophys.
413
17-22
2003
Lactococcus lactis
brenda
van Kuilenburg, A.B.P.; Elzinga, L.; Van Gennip, A.H.
Kinetic properties of CTP synthetase from HL-60 cells
Adv. Exp. Med. Biol.
431
255-258
1998
Homo sapiens
brenda
van Kuilenburg, A.B.P.; Meinsma, R.; Vreken, P.; Waterham, H.R.; van Gennip, A.H.
Isoforms of human CTP synthetase
Adv. Exp. Med. Biol.
486
257-261
2000
Homo sapiens
brenda
Meng, Q.; Switzer, R.L.
Regulation of transcription of the Bacillus subtilis pyrG gene, encoding cytidine triphosphate synthetase
J. Bacteriol.
183
5513-5522
2001
Bacillus subtilis
brenda
Hofer, A.; Steverding, D.; Chabes, A.; Brun, R.; Thelander, L.
Trypanosoma brucei CTP synthetase: a target for the treatment of African sleeping sickness
Proc. Natl. Acad. Sci. USA
98
6412-6416
2001
Trypanosoma brucei
brenda
Park, T.S.; O'Brien, D.J.; Carman, G.M.
Phosphorylation of CTP synthetase on Ser36, Ser330, Ser354, and Ser454 regulates the levels of CTP and phosphatidylcholine synthesis in Saccharomyces cerevisiae
J. Biol. Chem.
278
20785-20794
2003
Saccharomyces cerevisiae
brenda
Jorgensen, C.M.; Hammer, K.; Martinussen, J.
CTP limitation increases expression of CTP synthase in Lactococcus lactis
J. Bacteriol.
185
6562-6574
2003
Lactococcus lactis
brenda
Pappas, A.; Yang, W.L.; Park, T.S.; Carman, G.M.
Nucleotide-dependent tetramerization of CTP synthetase from Saccharomyces cerevisiae
J. Biol. Chem.
273
15954-15960
1998
Saccharomyces cerevisiae
brenda
Ostrander, D.B.; O'Brien, D.J.; Gorman, J.A.; Carman, G.M.
Effect of CTP synthetase regulation by CTP on phospholipid synthesis in Saccharomyces cerevisiae
J. Biol. Chem.
273
18992-19001
1998
Saccharomyces cerevisiae
brenda
Wadskov-Hansen, S.L.; Willemoes, M.; Martinussen, J.; Hammer, K.; Neuhard, J.; Larsen, S.
Cloning and verification of the Lactococcus lactis pyrG gene and characterization of the gene product, CTP synthase
J. Biol. Chem.
276
38002-38009
2001
Lactococcus lactis
brenda
Pappas, A.; Park, T.S.; Carman, G.M.
Characterization of a novel dUTP-dependent activity of CTP synthetase from Saccharomyces cerevisiae
Biochemistry
38
16671-16677
1999
Saccharomyces cerevisiae
brenda
Goto, M.; Omi, R.; Hoseki, J.; Nakagawa, N.; Miyahara, I.; Hirotsu, K.
Expression, purification and preliminary X-ray characterization of CTP synthetase from Thermus thermophilus HB8
Acta Crystallogr. Sect. D
59
551-553
2003
Thermus thermophilus, Thermus thermophilus HB8 / ATCC 27634 / DSM 579
brenda
Weng, M.; Zalkin, H.
Structural role for a conserved region in the CTP synthetase glutamine amide transfer domain
J. Bacteriol.
169
3023-3028
1987
Escherichia coli
brenda
Zalkin, H.
CTP synthetase
Methods Enzymol.
113
282-287
1985
Escherichia coli, Escherichia coli B / ATCC 11303
brenda
Kizaki, H.; Ohsaka, F.; Sakurada, T.
CTP synthetase from Ehrlich ascites tumor cells. Subunit stoichiometry and regulation of activity
Biochim. Biophys. Acta
829
34-43
1985
Mus musculus
brenda
von der Saal, W.; Villafranca, J.J.; Anderson, P.M.
Cytidine-5'-triphosphate synthetase catalyzes the phosphorylation of uridine 5'-triphosphate by adenosine 5'-triphosphate
J. Am. Chem. Soc.
107
703-704
1985
Chlamydia trachomatis, Escherichia coli
-
brenda
Anderson, P.M.
CTP synthetase from Escherichia coli: an improved purification procedure and characterization of hysteretic and enzyme concentration effects on kinetic properties
Biochemistry
22
3285-3292
1983
Escherichia coli
brenda
Kizaki, H.; Ohsaka, F.; Sakurada, T.
Role of GTP in CTP synthetase from Ehrlich ascites tumor cells
Biochem. Biophys. Res. Commun.
108
286-291
1982
Mus musculus
brenda
Scheit, K.H.; Linke, H.J.
Substrate specificity of CTP synthetase from Escherichia coli
Eur. J. Biochem.
126
57-60
1982
Escherichia coli
brenda
Kizaki, H.; Sakurada, T.; Weber, G.
Purification and properties of CTP synthetase from Ehrlich ascites tumor cells
Biochim. Biophys. Acta
662
48-54
1981
Mus musculus
brenda
Weinfeld, H.; Savage, C.R.; McPartland, R.P.
CTP synthetase of bovine calf liver
Methods Enzymol.
51
84-90
1978
Bos taurus
brenda
Long, C.; Koshland jr., D.E.
Cytidine triphosphate synthetase
Methods Enzymol.
51
79-83
1978
Escherichia coli
brenda
Williams, J.C.; Kizaki, H.; Weber, G.; Morris, H.P.
Increased CTP synthetase activity in cancer cells
Nature
271
71-73
1978
Rattus norvegicus
brenda
McPartland, R.P.; Weinfeld, H.
Cytidine 5'-triphosphate synthetase of calf liver
J. Biol. Chem.
251
4372-4378
1976
Bos taurus
brenda
Koshland jr., D.E.; Levitzki, A.
CTP synthetase and related enzymes
The Enzymes, 3rd Ed. (Boyer, P. D. , ed. )
10
539-559
1974
Escherichia coli
-
brenda
Scheit, K.H.; Linke, H.J.
Substrate specificity of CTP synthetase from E. coli
Nucleic Acids Res.
9
229-233
1981
Escherichia coli
brenda
Yang, W.L.; Carman, G.M.
Phosphorylation and regulation of CTP synthetase from Saccharomyces cerevisiae by protein kinase A
J. Biol. Chem.
271
28777-28783
1996
Saccharomyces cerevisiae
brenda
Wylie, J.L.; Wang, L.L.; Tipples, G.; McClarty, G.
A single point mutation in CTP synthetase of Chlamydia trachomatis confers resistance to cyclopentenyl cytosine
J. Biol. Chem.
271
15393-15400
1996
Chlamydia trachomatis, Chlamydia trachomatis L2/CPEC
brenda
Kuilenburg, A.B.P.; Elzinga, L.; van den Berg, A.A.; Slingerland, R.J.; van Gennip, A.H.
A fast and novel assay of CTP synthetase. Evidence for hysteretic properties of the mammalian enzyme
Anticancer Res.
14
411-416
1994
Bos taurus
brenda
Lewis, D.A.; Villafranca, J.J.
Investigation of the mechanism of CTP synthetase using rapid quench and isotope partitioning methods
Biochemistry
28
8454-8459
1989
Escherichia coli
brenda
Kang, G.J.; Cooney, D.A.; Moyer, J.D.; Kelley, J.A.; Kim, H.Y.; Marquez, V.E.; Johns, D.G.
Cyclopentenylcytosine triphosphate. Formation and inhibition of CTP synthetase
J. Biol. Chem.
264
713-718
1989
Mus musculus
brenda
Yang, W.L.; McDonough, V.M.; Ozier-Kalogeropoulos, O.; Adeline, M.T.; Flocco, M.T.; Carman, G.M.
Purification and characterization of CTP synthetase, the product of the URA7 gene in Saccharomyces cerevisiae
Biochemistry
33
10785-10793
1994
Saccharomyces cerevisiae
brenda
Wylie, J.L.; Berry, J.D.; McClarty, G.
Chlamydia trachomatis CTP synthetase: molecular characterization and developmental regulation of expression
Mol. Microbiol.
22
631-642
1996
Chlamydia trachomatis, Escherichia coli
brenda
Aronow, B.; Ullman, B.
In situ regulation of mammalian CTP synthetase by allosteric inhibition
J. Biol. Chem.
262
5106-5112
1987
Mus musculus
brenda
Robertson, J.G.; Villafranca, J.J.
Characterization of metal ion activation and inhibition of CTP synthetase
Biochemistry
32
3769-3777
1993
Escherichia coli
brenda
Kizaki, H.; Ohsaka, F.; Sakurada, T.
Synthesis of N4-substituted CTP by mammalian CTP synthetase
Biochem. Biophys. Res. Commun.
145
569-574
1987
Mus musculus
brenda
Willems, M.
Competition between ammonia derived from internal glutamine hydrolysis and hydroxylamine present in the solution for incorporation into UTP as catalysed by Lactococcus lactis CTP synthase
Arch. Biochem. Biophys.
424
105-111
2004
Lactococcus lactis
brenda
Endrizzi, J.A.; Kim, H.; Anderson, P.M.; Baldwin, E.P.
Crystal structure of Escherichia coli cytidine triphosphate synthetase, a nucleotide-regulated glutamine amidotransferase/ATP-dependent amidoligase fusion protein and homologue of anticancer and antiparasitic drug targets
Biochemistry
43
6447-6463
2004
Escherichia coli (P0A7E5), Escherichia coli
brenda
MacDonnell, J.E.; Lunn, F.A.; Bearne, S.L.
Inhibition of E. coli CTP synthase by the "positive" allosteric effector GTP
Biochim. Biophys. Acta
1699
213-220
2004
Escherichia coli
brenda
Lunn, F.A.; Bearne, S.L.
Alternative substrates for wild-type and L109A E. coli CTP synthases: kinetic evidence for a constricted ammonia tunnel
Eur. J. Biochem.
271
4204-4212
2004
Escherichia coli (P0A7E5), Escherichia coli
brenda
Willems, M.; Molgaard, A.; Johansson, E.; Martinussen, J.
Lid L11 of the glutamine amidotransferase domain of CTP synthase mediates allosteric GTP activation of glutaminase activity
FEBS J.
272
856-864
2005
Lactococcus lactis
brenda
Yuan, P.; Hendriks, E.F.; Fernandez, H.R.; O'Sullivan, W.J.; Stewart, T.S.
Functional expression of the gene encoding cytidine triphosphate synthetase from Plasmodium falciparum which contains two novel sequences that are potential antimalarial targets
Mol. Biochem. Parasitol.
143
200-208
2005
Plasmodium falciparum
brenda
Goto, M.; Omi, R.; Nakagawa, N.; Miyahara, I.; Hirotsu, K.
Crystal structures of CTP synthetase reveal ATP, UTP, and glutamine binding sites
Structure
12
1413-1423
2004
Thermus thermophilus HB8 (Q5SIA8), Thermus thermophilus HB8
brenda
Kursula, P.; Flodin, S.; Ehn, M.; Hammarstroem, M.; Schueler, H.; Nordlund, P.; Stenmark, P.
Structure of the synthetase domain of human CTP synthetase, a target for anticancer therapy
Acta Crystallogr. Sect. F
62
613-617
2006
Homo sapiens
brenda
MacLeod, T.J.; Lunn, F.A.; Bearne, S.L.
The role of lysine residues 297 and 306 in nucleoside triphosphate regulation of E. coli CTP synthase: inactivation by 2,3-dialdehyde ATP and mutational analyses
Biochim. Biophys. Acta
1764
199-210
2006
Escherichia coli (A0A140N932), Escherichia coli
brenda
Fijolek, A.; Hofer, A.; Thelander, L.
Expression, purification, characterization, and in vivo targeting of trypanosome CTP synthetase for treatment of African sleeping sickness
J. Biol. Chem.
282
11858-11865
2007
Trypanosoma brucei
brenda
Chang, Y.F.; Martin, S.S.; Baldwin, E.P.; Carman, G.M.
Phosphorylation of human CTP synthetase 1 by protein kinase C: identification of Ser(462) and Thr(455) as major sites of phosphorylation
J. Biol. Chem.
282
17613-17622
2007
Homo sapiens
brenda
Higgins, M.J.; Graves, P.R.; Graves, L.M.
Regulation of human cytidine triphosphate synthetase 1 by glycogen synthase kinase 3
J. Biol. Chem.
282
29493-29503
2007
Homo sapiens (Q9NRF8), Homo sapiens
brenda
Choi, M.G.; Carman, G.M.
Phosphorylation of human CTP synthetase 1 by protein kinase A: identification of Thr455 as a major site of phosphorylation
J. Biol. Chem.
282
5367-5377
2007
Homo sapiens
brenda
Lunn, F.A.; Macdonnell, J.E.; Bearne, S.L.
Structural requirements for the activation of Escherichia coli CTP synthase by the allosteric effector GTP are stringent, but requirements for inhibition are Lax
J. Biol. Chem.
283
2010-2020
2007
Escherichia coli (P0A7E5), Escherichia coli
brenda
Braun, O.; Knipp, M.; Chesnov, S.; Vasak, M.
Specific reactions of S-nitrosothiols with cysteine hydrolases: A comparative study between dimethylargininase-1 and CTP synthetase
Protein Sci.
16
1522-1534
2007
Escherichia coli (P0A7E5)
brenda
Lunn, F.A.; Macleod, T.J.; Bearne, S.L.
Mutational analysis of conserved glycine residues 142, 143 and 146 reveals Gly(142) is critical for tetramerization of CTP synthase from Escherichia coli
Biochem. J.
412
113-121
2008
Escherichia coli (P0A7E5), Escherichia coli
brenda
Higgins, M.J.; Loiselle, D.; Haystead, T.A.; Graves, L.M.
Human cytidine triphosphate synthetase 1 interacting proteins
Nucleosides Nucleotides Nucleic Acids
27
850-857
2008
Homo sapiens
brenda
Chang, Y.F.; Carman, G.M.
CTP synthetase and its role in phospholipid synthesis in the yeast Saccharomyces cerevisiae
Prog. Lipid Res.
47
333-339
2008
Saccharomyces cerevisiae, Homo sapiens
brenda
Roy, A.C.; Lunn, F.A.; Bearne, S.L.
Inhibition of CTP synthase from Escherichia coli by xanthines and uric acids
Bioorg. Med. Chem. Lett.
20
141-144
2010
Escherichia coli
brenda
Lauritsen, I.; Willemoes, M.; Jensen, K.F.; Johansson, E.; Harris, P.
Structure of the dimeric form of CTP synthase from Sulfolobus solfataricus
Acta Crystallogr. Sect. F
67
201-208
2011
Saccharolobus solfataricus (Q980S6), Saccharolobus solfataricus
brenda
Kassel, K.M.; Au, d.a..R.; Higgins, M.J.; Hines, M.; Graves, L.M.
Regulation of human cytidine triphosphate synthetase 2 by phosphorylation
J. Biol. Chem.
285
33727-33736
2010
Homo sapiens, Homo sapiens (Q9NRF8)
brenda
Liu, J.L.
Intracellular compartmentation of CTP synthase in Drosophila
J. Genet. Genomics
37
281-296
2010
Drosophila melanogaster
brenda
Ingerson-Mahar, M.; Briegel, A.; Werner, J.N.; Jensen, G.J.; Gitai, Z.
The metabolic enzyme CTP synthase forms cytoskeletal filaments
Nat. Cell Biol.
12
739-746
2010
Caulobacter vibrioides, Escherichia coli
brenda
Steeves, C.H.; Bearne, S.L.
Activation and inhibition of CTP synthase from Trypanosoma brucei, the causative agent of African sleeping sickness
Bioorg. Med. Chem. Lett.
21
5188-5190
2011
Trypanosoma brucei
brenda
Gou, K.M.; Chang, C.C.; Shen, Q.J.; Sung, L.Y.; Liu, J.L.
CTP synthase forms cytoophidia in the cytoplasm and nucleus
Exp. Cell Res.
323
242-253
2014
Homo sapiens, Mus musculus
brenda
Azzam, G.; Liu, J.L.
Only one isoform of Drosophila melanogaster CTP synthase forms the cytoophidium
PLoS Genet.
9
e1003256
2013
Drosophila melanogaster
brenda
Strochlic, T.I.; Stavrides, K.P.; Thomas, S.V.; Nicolas, E.; OReilly, A.M.; Peterson, J.R.
Ack kinase regulates CTP synthase filaments during Drosophila oogenesis
EMBO Rep.
15
1184-1191
2014
Drosophila melanogaster (X2JCY2)
brenda
Huang, Y.; Wang, J.J.; Ghosh, S.; Liu, J.L.
Critical roles of CTP synthase N-terminal in cytoophidium assembly
Exp. Cell Res.
354
122-133
2017
Drosophila melanogaster (X2JCY2)
brenda
Chang, C.C.; Jeng, Y.M.; Peng, M.; Keppeke, G.D.; Sung, L.Y.; Liu, J.L.
CTP synthase forms the cytoophidium in human hepatocellular carcinoma
Exp. Cell Res.
361
292-299
2017
Homo sapiens, Drosophila melanogaster (X2JCY2)
brenda
Wang, P.Y.; Lin, W.C.; Tsai, Y.C.; Cheng, M.L.; Lin, Y.H.; Tseng, S.H.; Chakraborty, A.; Pai, L.M.
Regulation of CTP synthase filament formation during DNA endoreplication in Drosophila
Genetics
201
1511-1523
2015
Drosophila melanogaster (X2JCY2)
brenda
Li, X.; Xie, J.; Hei, M.; Tang, J.; Wang, Y.; Foerster, E.; Zhao, S.
High level of CTP synthase induces formation of cytoophidia in cortical neurons and impairs corticogenesis
Histochem. Cell Biol.
149
61-73
2018
Mus musculus
brenda
Noree, C.; Monfort, E.; Shiau, A.K.; Wilhelm, J.E.
Common regulatory control of CTP synthase enzyme activity and filament formation
Mol. Biol. Cell
25
2282-2290
2014
Saccharomyces cerevisiae (P28274), Saccharomyces cerevisiae, Saccharomyces cerevisiae ATCC 204508 (P28274)
brenda
Aughey, G.; Grice, S.; Liu, J.
The Interplay between Myc and CTP synthase in Drosophila
PLoS Genet.
12
e1005867
2016
Drosophila melanogaster (X2JCY2)
brenda
Sandalli, C.; Saral, A.; Uelker, S.; Karaoglu, H.; Belduez, A.; Copur Cicek, A.
Cloning, expression, and characterization of a novel CTP synthase gene from Anoxybacillus gonensis G2
Turk. J. Biol.
38
111-117
2014
Anoxybacillus gonensis (A0A0A2SLZ4), Anoxybacillus gonensis G2 (A0A0A2SLZ4)
-
brenda
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