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Literature summary for 6.3.4.2 extracted from

  • 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 (2007), J. Biol. Chem., 283, 2010-2020.
    View publication on PubMed
Search for more literature for 6.3.4.2

Activating Compound

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

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(Commentary)

Cloned (Comment) Organism
cloned in Escherichia coli as a histidine-tagged fusion protein Escherichia coli

Inhibitors

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 [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information kinetic mechanism, activation and inhibition kinetics Escherichia coli

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+
-
 Escherichia coli
Mg2+ assay with 10 mM MgCl2 Escherichia coli

Natural Substrates/ Products (Substrates)

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

Organism UniProt Comment Textmining
Escherichia coli P0A7E5
-
-

Purification (Commentary)

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 [micromol/min/mg]

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 and Products (Substrate)

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

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 [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Escherichia coli

Turnover Number [1/s]

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

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
 assay at Escherichia coli

Cofactor

Cofactor Comment Organism Structure
ATP dependent on Escherichia coli

Ki Value [mM]

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 : 0.2–0.5 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 : 4–12 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 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