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Literature summary for 6.3.4.2 extracted from

  • 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 (2006), Biochim. Biophys. Acta, 1764, 199-210.
    View publication on PubMed
Search for more literature for 6.3.4.2

Cloned(Commentary)

Cloned (Comment) Organism
cloned in Escherichia coli BL-21 as an N-terminal 6xHis-tag fusion protein Escherichia coli

Protein Variants

Protein Variants Comment Organism
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 Escherichia coli
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 Escherichia coli
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 Escherichia coli

Inhibitors

Inhibitors Comment Organism Structure
2',3'-dialdehyde adenosine 5'-triphosphate irreversible inhibitor of CTPS Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.196
-
 glutamine mutant K306A, presence of 1 mM ATP, 1 mM UTP Escherichia coli
0.206
-
 glutamine wild-type protein, presence of 3 mM ATP, 2 mM UTP Escherichia coli
0.345
-
 glutamine wild-type protein, presence of 1 mM ATP, 1 mM UTP Escherichia coli
0.424
-
 glutamine mutant K306A, presence of 3 mM ATP, 2 mM UTP Escherichia coli
0.627
-
 NH3 mutant K306A, presence of 1 mM ATP, 1 mM UTP Escherichia coli
1.47
-
 NH3 mutant K306A, presence of 3 mM ATP, 2 mM UTP Escherichia coli
1.54
-
 NH3 mutant K297A, presence of 1 mM ATP, 1 mM UTP Escherichia coli
2.15
-
 NH3 wild-type protein, presence of 1 mM ATP, 1 mM UTP Escherichia coli
2.79
-
 NH3 wild-type protein, presence of 3 mM ATP, 2 mM UTP Escherichia coli

Metals/Ions

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

Organism

Organism UniProt Comment Textmining
Escherichia coli A0A140N932 BL21(DE3)
-

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 + NH3
-
 Escherichia coli ADP + CTP + phosphate
-
 ?

Synonyms

Synonyms Comment Organism
CTPS
-
 Escherichia coli
cytidine 5'-triphosphate synthetase
-
 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
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 Escherichia coli
0.031 0.51 NH3 mutant K306A, presence of 3 mM ATP, 2 mM UTP Escherichia coli
0.054
-
 2',3'-dialdehyde adenosine 5'-triphosphate in the presence of 10 mM UTP Escherichia coli
1.28
-
 glutamine mutant K306A, presence of 1 mM ATP, 1 mM UTP Escherichia coli
1.35
-
 glutamine mutant K306A, presence of 3 mM ATP, 2 mM UTP Escherichia coli
1.5
-
 glutamine wild-type protein, presence of 3 mM ATP, 2 mM UTP Escherichia coli
2.19
-
 NH3 mutant K306A, presence of 1 mM ATP, 1 mM UTP Escherichia coli
6.1
-
 glutamine wild-type protein, presence of 1 mM ATP, 1 mM UTP Escherichia coli
6.26
-
 ATP K306A, presence of 2 mM UTP Escherichia coli
6.9
-
 UTP K306A, presence of 2 mM ATP Escherichia coli
7.59
-
 NH3 mutant K306A, presence of 3 mM ATP, 2 mM UTP Escherichia coli
8.59
-
 NH3 mutant K297A, presence of 1 mM ATP, 1 mM UTP Escherichia coli
9.5
-
 NH3 wild-type protein, presence of 1 mM ATP, 1 mM UTP Escherichia coli
10.8
-
 ATP K297A, presence of 2 mM UTP Escherichia coli
11.3
-
 NH3 wild-type protein, presence of 3 mM ATP, 2 mM UTP Escherichia coli
12.8
-
 ATP wild-type protein, presence of 2 mM UTP Escherichia coli
13.7
-
 UTP wild-type protein, presence of 3 mM ATP Escherichia coli
14
-
 UTP K297A, presence of 3 mM ATP Escherichia coli

pH Optimum

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

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
3.36
-
 2',3'-dialdehyde adenosine 5'-triphosphate in the presence of 10 mM UTP Escherichia coli
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 Escherichia coli