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Literature summary for 6.3.5.5 extracted from

  • Hart, E.J.; Powers-Lee, S.G.
    Mutation analysis of carbamoyl phosphate synthetase: does the structurally conserved glutamine amidotransferase triad act as a functional dyad? (2008), Protein Sci., 17, 1120-1128.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expression of wild-type and mutant enzymes in Escherichia coli strain L673 Escherichia coli

Protein Variants

Protein Variants Comment Organism
D334A site-directed mutagenesis, the mutation has essentially no effect on the Km for L-glutamine Escherichia coli
N311A site-directed mutagenesis, the mutant shows increased Km for L-glutamine compared to the wild-type enzyme Escherichia coli
Q310A site-directed mutagenesis, the mutant shows increased Km for L-glutamine compared to the wild-type enzyme Escherichia coli
Q351A site-directed mutagenesis, the mutant shows highly increased Km for L-glutamine compared to the wild-type enzyme Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information kinetics of wild-type and mutant enzymes in L-glutamine-dependent and bicarbonate-dependent ATPase activities, overview Escherichia coli
0.027
-
ATP pH 7.6, 25°C, mutant D334A, L-glutamine hydrolysis Escherichia coli
0.044
-
ATP pH 7.6, 25°C, wild-type enzyme, L-glutamine hydrolysis Escherichia coli
0.052
-
ATP pH 7.6, 25°C, mutant Q351A, L-glutamine hydrolysis Escherichia coli
0.067
-
ATP pH 7.6, 25°C, mutant Q310A, L-glutamine hydrolysis Escherichia coli
0.075
-
ATP pH 7.6, 25°C, mutant N311A, L-glutamine hydrolysis Escherichia coli
0.13
-
L-glutamine pH 7.6, 25°C, wild-type enzyme, L-glutamine hydrolysis Escherichia coli
0.15
-
L-glutamine pH 7.6, 25°C, mutant Q351A, L-glutamine hydrolysis Escherichia coli
1.99
-
L-glutamine pH 7.6, 25°C, mutant Q310A, L-glutamine hydrolysis Escherichia coli
3.45
-
L-glutamine pH 7.6, 25°C, mutant N311A, L-glutamine hydrolysis Escherichia coli
18.87
-
L-glutamine pH 7.6, 25°C, mutant D334A, L-glutamine hydrolysis Escherichia coli

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+
-
Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + L-glutamine + HCO3- + H2O Escherichia coli
-
ADP + phosphate + L-glutamate + carbamoyl phosphate
-
?

Organism

Organism UniProt Comment Textmining
Escherichia coli
-
-
-

Reaction

Reaction Comment Organism Reaction ID
2 ATP + L-glutamine + hydrogencarbonate + H2O = 2 ADP + phosphate + L-glutamate + carbamoyl phosphate mechanism for the generation of ammonia within the L-glutamine amidotransferase domain of CPS, detailed overview Escherichia coli

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
-
Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + L-glutamine + HCO3- + H2O
-
Escherichia coli ADP + phosphate + L-glutamate + carbamoyl phosphate
-
?
ATP + L-glutamine + HCO3- + H2O evolutionarily conserved triad glutamine amidotransferase, GAT, domains catalyze the cleavage of L-glutamine to yield ammonia and sequester the ammonia in a tunnel until delivery to a variety of acceptor substrates in synthetase domains of variable structure. The Cys269 and His353 catalytic triad residues are essential for L-glutamine hydrolysis, whereas Glu355 is not critical for eCPS activity, Gln351 plays a key role in L-glutamine binding, overview Escherichia coli ADP + phosphate + L-glutamate + carbamoyl phosphate
-
?

Synonyms

Synonyms Comment Organism
carbamoyl phosphate synthetase
-
Escherichia coli
CPS
-
Escherichia coli

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.53
-
L-glutamine pH 7.6, 25°C, mutant Q351A, L-glutamine hydrolysis Escherichia coli
1.05
-
L-glutamine pH 7.6, 25°C, mutant N311A, L-glutamine hydrolysis Escherichia coli
1.17
-
L-glutamine pH 7.6, 25°C, mutant D334A, L-glutamine hydrolysis Escherichia coli
1.33
-
ATP pH 7.6, 25°C, mutant D334A, L-glutamine hydrolysis Escherichia coli
1.6
-
L-glutamine pH 7.6, 25°C, mutant Q310A, L-glutamine hydrolysis Escherichia coli
2.13
-
ATP pH 7.6, 25°C, mutant N311A, L-glutamine hydrolysis Escherichia coli
2.13
-
L-glutamine pH 7.6, 25°C, wild-type enzyme, L-glutamine hydrolysis Escherichia coli
3.19
-
ATP pH 7.6, 25°C, mutant Q351A, L-glutamine hydrolysis Escherichia coli
4.79
-
ATP pH 7.6, 25°C, wild-type enzyme, L-glutamine hydrolysis Escherichia coli
5.75
-
ATP pH 7.6, 25°C, mutant Q310A, L-glutamine hydrolysis Escherichia coli

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.6
-
assay at Escherichia coli

Cofactor

Cofactor Comment Organism Structure
ATP
-
Escherichia coli