Activating Compound | Comment | Organism | Structure |
---|---|---|---|
2'-deoxy-GTP | 0 mM guanosine, kact: 1.5/sec, KA: 0.21 mM, Ki: 0.36 mM | Escherichia coli | |
2'-deoxy-GTP | the GTP analogue is capable of activating Gln-dependent CTP formation | Escherichia coli | |
6-thio-GTP | the GTP analogue is capable of activating Gln-dependent CTP formation | Escherichia coli | |
6-thioguanosine 5'-triphosphate | 0 mM guanosine, kact: 8.5/sec, KA: 0.035 mM, Ki: 0.27 mM | Escherichia coli | |
GTP | 0 mM guanosine, kact: 10.6/sec, KA: 0.081 mM, Ki: 0.28 mM | Escherichia coli | |
GTP | 0.1 mM guanosine, kact: 10.3/sec, KA: 0.088 mM, Ki: 0.22 mM | Escherichia coli | |
GTP | 0.2 mM guanosine, kact: 8.2/sec, KA: 0.078 mM, Ki: 0.12 mM | Escherichia coli | |
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 | Escherichia coli | |
guanosine 5'-tetraphosphate | 0 mM guanosine, kact: 4/sec, KA: 0.19 mM, Ki: 0.5 mM | Escherichia coli | |
guanosine 5'-tetraphosphate | the GTP analogue is capable of activating Gln-dependent CTP formation | Escherichia coli | |
ITP | 0 mM guanosine, kact: 5.2/sec, KA: 2.9 mM, Ki: 4.5 mM | Escherichia coli | |
ITP | the GTP analogue is capable of activating Gln-dependent CTP formation | Escherichia coli | |
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 | Escherichia coli | |
O-methylguanosine 5'-triphosphate | 0 mM guanosine, kact: 2.8/sec, KA: 0.13 mM, Ki: 0.29 mM | Escherichia coli | |
O6-methyl-GTP | the GTP analogue is capable of activating Gln-dependent CTP formation | Escherichia coli |
Application | Comment | Organism |
---|---|---|
drug development | modifying 2-aminopurine or 2-aminopurine riboside may serve as an effective strategy for developing CTPS inhibitors | Escherichia coli |
Cloned (Comment) | Organism |
---|---|
cloned in Escherichia coli as a histidine-tagged fusion protein | Escherichia coli |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
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 | Escherichia coli | |
2'-deoxy-guanosine | - |
Escherichia coli | |
2,6-diaminopurine riboside | - |
Escherichia coli | |
2-aminopurine riboside | - |
Escherichia coli | |
3'-deoxy-guanosine | - |
Escherichia coli | |
6-thio-GTP | the GTP analogue is capable of inhibiting Gln-dependent CTP formation at over 0.15 mM | Escherichia coli | |
6-thioguanine | - |
Escherichia coli | |
6-thioguanosine | - |
Escherichia coli | |
6-thioguanosine 5'-triphosphate | no guanosine, kact: 8.5/sec, KA: 0.035 mM, Ki: 0.27 mM | Escherichia coli | |
7-deazaguanosine | - |
Escherichia coli | |
8-oxoguanosine | - |
Escherichia coli | |
8-oxoguanosine 5'-triphosphate | - |
Escherichia coli | |
acycloguanosine | - |
Escherichia coli | |
acycloguanosine monophosphate | - |
Escherichia coli | |
adenosine | - |
Escherichia coli | |
dideoxy-GTP | - |
Escherichia coli | |
GDP | - |
Escherichia coli | |
GMP | - |
Escherichia coli | |
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 | Escherichia coli | |
guanosine | - |
Escherichia coli | |
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 | Escherichia coli | |
Inosine | - |
Escherichia coli | |
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 | Escherichia coli | |
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 | Escherichia coli | |
N-methylguanosine | - |
Escherichia coli | |
O-methylguanosine | - |
Escherichia coli | |
O-methylguanosine 5'-triphosphate | no guanosine, kact: 2.8/sec, KA: 0.13 mM, Ki: 0.29 mM | Escherichia coli | |
O6-methyl-GTP | the GTP analogue is capable of inhibiting Gln-dependent CTP formation at over 0.15 mM | Escherichia coli | |
Xanthosine | - |
Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetic mechanism, activation and inhibition kinetics | Escherichia coli |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | - |
Escherichia coli | |
Mg2+ | assay with 10 mM MgCl2 | Escherichia coli |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + UTP + L-glutamine | Escherichia coli | - |
ADP + phosphate + CTP + L-glutamate | - |
? | |
ATP + UTP + NH3 | Escherichia coli | - |
ADP + phosphate + CTP | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P0A7E5 | - |
- |
Purification (Comment) | Organism |
---|---|
the soluble histidine-tagged CTPS is purified using metal ion affinity chromatography and the histidine tag is subsequently removed using thrombin-catalyzed cleavage | Escherichia coli |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
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. | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + UTP + Gln + H2O | - |
Escherichia coli | ADP + CTP + Glu + phosphate | - |
? | |
ATP + UTP + L-glutamine | - |
Escherichia coli | ADP + phosphate + CTP + L-glutamate | - |
? | |
ATP + UTP + NH3 | - |
Escherichia coli | ADP + CTP + phosphate | - |
? | |
ATP + UTP + NH3 | - |
Escherichia coli | ADP + phosphate + CTP | - |
? | |
deoxyGTP + UTP + NH4+ | - |
Escherichia coli | deoxyGDP + phosphate + CTP | - |
? | |
GTP + UTP + NH4+ | - |
Escherichia coli | GDP + phosphate + CTP | - |
? |
Synonyms | Comment | Organism |
---|---|---|
CTP synthase | - |
Escherichia coli |
CTPS | - |
Escherichia coli |
cytidine 5'-triphosphate synthase | - |
Escherichia coli |
UTP:ammonia ligase (ADP-forming) | - |
Escherichia coli |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.5 | - |
2'-deoxy-GTP | no guanosine | Escherichia coli | |
2.8 | - |
O-methylguanosine 5'-triphosphate | no guanosine | Escherichia coli | |
4 | - |
guanosine 5'-tetraphosphate | no guanosine | Escherichia coli | |
5.2 | - |
ITP | no guanosine | Escherichia coli | |
8.2 | - |
GTP | 0.2 mM guanosine | Escherichia coli | |
8.5 | - |
6-thioguanosine 5'-triphosphate | no guanosine, kact: 8.5/sec, KA: 0.035 mM, Ki: 0.27 mM | Escherichia coli | |
10.3 | - |
GTP | 0.1 mM guanosine | Escherichia coli | |
10.6 | - |
GTP | no guanosine | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Escherichia coli |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | dependent on | Escherichia coli |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
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). | Escherichia coli | |
additional information | - |
additional information | kinetic mechanism, activation and inhibition kinetics | Escherichia coli | |
0.12 | - |
GTP | 0.2 mM guanosine | Escherichia coli | |
0.22 | - |
GTP | 0.1 mM guanosine | Escherichia coli | |
0.27 | - |
6-thioguanosine 5'-triphosphate | - |
Escherichia coli | |
0.28 | - |
GTP | no guanosine | Escherichia coli | |
0.29 | - |
O-methylguanosine 5'-triphosphate | - |
Escherichia coli | |
0.36 | - |
2'-deoxy-GTP | - |
Escherichia coli | |
0.5 | - |
guanosine 5'-tetraphosphate | - |
Escherichia coli | |
4.5 | - |
ITP | - |
Escherichia coli |
IC50 Value | IC50 Value Maximum | Comment | Organism | Inhibitor | Structure |
---|---|---|---|---|---|
0.08 | - |
- |
Escherichia coli | 8-oxoguanosine | |
0.08 | - |
IC50 for inhibition of Gln-dependent CTP formation | Escherichia coli | 8-oxoguanosine | |
0.11 | - |
IC50 for inhibition of Gln-dependent CTP formation | Escherichia coli | 2,6-diaminopurine riboside | |
0.11 | - |
IC50 for inhibition of Gln-dependent CTP formation | Escherichia coli | 8-oxoguanosine 5'-triphosphate | |
0.13 | - |
IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | 8-oxoguanosine | |
0.15 | - |
0.30 mM Gln, IC50 for inhibition of Gln-dependent CTP formation | Escherichia coli | O-methylguanosine | |
0.15 | - |
IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | 8-oxoguanosine 5'-triphosphate | |
0.16 | - |
10 mM Gln, IC50 for inhibition of Gln-dependent CTP formation | Escherichia coli | O-methylguanosine | |
0.16 | - |
2.5 mM Gln, IC50 for inhibition of Gln-dependent CTP formation | Escherichia coli | O-methylguanosine | |
0.16 | - |
IC50 for inhibition of Gln-dependent CTP formation | Escherichia coli | O-methylguanosine | |
0.17 | - |
0.75 mM Gln, IC50 for inhibition of Gln-dependent CTP formation | Escherichia coli | O-methylguanosine | |
0.17 | - |
IC50 for inhibition of Gln-dependent CTP formation | Escherichia coli | N-methylguanosine | |
0.18 | - |
IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | 2,6-diaminopurine riboside | |
0.22 | - |
5.44 mM NH3, IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | guanosine | |
0.22 | - |
IC50 for inhibition of Gln-dependent CTP formation | Escherichia coli | Xanthosine | |
0.22 | - |
IC50 for inhibition of Gln-dependent CTP formation | Escherichia coli | 2-aminopurine riboside | |
0.23 | - |
5.44 mM NH3, IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | GMP | |
0.23 | - |
IC50 for inhibition of Gln-dependent CTP formation | Escherichia coli | 6-thioguanosine | |
0.23 | - |
IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | N-methylguanosine | |
0.25 | - |
2.72 mM NH3, IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | GMP | |
0.25 | - |
IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | O-methylguanosine | |
0.26 | - |
1.36 mM NH3, IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | guanosine | |
0.26 | - |
2.72 mM NH3, IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | guanosine | |
0.26 | - |
IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | GMP | |
0.26 | - |
IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | 2-aminopurine riboside | |
0.28 | - |
1.36 mM NH3, IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | GMP | |
0.29 | - |
2.72 mM NH3, IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | GDP | |
0.29 | - |
5.44 mM NH3, IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | GTP | |
0.29 | - |
5.44 mM NH3, IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | GDP | |
0.29 | - |
8.16 mM NH3, IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | GMP | |
0.29 | - |
8.16 mM NH3, IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | guanosine | |
0.29 | - |
IC50 for inhibition of Gln-dependent CTP formation | Escherichia coli | dideoxy-GTP | |
0.29 | - |
IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | GDP | |
0.29 | - |
IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | guanosine | |
0.29 | - |
IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | Xanthosine | |
0.3 | - |
0.30 mM Gln, IC50 for inhibition of Gln-dependent CTP formation | Escherichia coli | guanosine | |
0.3 | - |
1.36 mM NH3, IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | GDP | |
0.3 | - |
2.72 mM NH3, IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | GTP | |
0.3 | - |
8.16 mM NH3, IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | GTP | |
0.3 | - |
IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | GTP | |
0.31 | - |
1.36 mM NH3, IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | GTP | |
0.31 | - |
IC50 for inhibition of Gln-dependent CTP formation | Escherichia coli | acycloguanosine monophosphate | |
0.32 | - |
10 mM Gln, IC50 for inhibition of Gln-dependent CTP formation | Escherichia coli | guanosine | |
0.32 | - |
2.5 mM Gln, IC50 for inhibition of Gln-dependent CTP formation | Escherichia coli | guanosine | |
0.32 | - |
IC50 for inhibition of Gln-dependent CTP formation | Escherichia coli | guanosine | |
0.33 | - |
IC50 for inhibition of Gln-dependent CTP formation | Escherichia coli | GDP | |
0.33 | - |
IC50 for inhibition of Gln-dependent CTP formation | Escherichia coli | GMP | |
0.33 | - |
IC50 for inhibition of Gln-dependent CTP formation | Escherichia coli | guanosine 5'-tetraphosphate | |
0.33 | - |
IC50 for inhibition of Gln-dependent CTP formation | Escherichia coli | acycloguanosine | |
0.33 | - |
IC50 for inhibition of Gln-dependent CTP formation | Escherichia coli | 2'-deoxy-GTP | |
0.33 | - |
IC50 for inhibition of Gln-dependent CTP formation | Escherichia coli | 2'-deoxy-guanosine | |
0.34 | - |
IC50 for inhibition of Gln-dependent CTP formation | Escherichia coli | 3'-deoxy-guanosine | |
0.35 | - |
0.75 mM Gln, IC50 for inhibition of Gln-dependent CTP formation | Escherichia coli | guanosine | |
0.39 | - |
IC50 for inhibition of Gln-dependent CTP formation | Escherichia coli | 6-thioguanine | |
0.4 | - |
IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | 6-thioguanosine | |
0.4 | - |
IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | 3'-deoxy-guanosine | |
0.41 | - |
IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | dideoxy-GTP | |
0.41 | - |
IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | acycloguanosine monophosphate | |
0.42 | - |
IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | guanosine 5'-tetraphosphate | |
0.42 | - |
IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | 2'-deoxy-GTP | |
0.45 | - |
8.16 mM NH3, IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | acycloguanosine | |
0.45 | - |
IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | acycloguanosine | |
0.45 | - |
IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | 2'-deoxy-guanosine | |
0.47 | - |
5.44 mM NH3, IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | acycloguanosine | |
0.48 | - |
2.72 mM NH3, IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | acycloguanosine | |
0.49 | - |
1.36 mM NH3, IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | acycloguanosine | |
0.61 | - |
IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | 6-thioguanine | |
2.9 | - |
8.16 mM NH3, IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | ITP | |
3 | - |
2.72 mM NH3, IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | ITP | |
3.1 | - |
5.44 mM NH3, IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | ITP | |
3.3 | - |
1.36 mM NH3, IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | ITP | |
3.5 | - |
IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | Inosine | |
3.7 | - |
IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | ITP | |
4.1 | - |
IC50 for inhibition of Gln-dependent CTP formation | Escherichia coli | ITP | |
5.2 | - |
IC50 for inhibition of Gln-dependent CTP formation | Escherichia coli | Inosine | |
11 | - |
IC50 for inhibition of Gln-dependent CTP formation | Escherichia coli | adenosine | |
12 | - |
IC50 for inhibition of NH3-dependent CTP formation | Escherichia coli | adenosine |