Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 6.3.5.5 extracted from

  • Fan, Y.; Lund, L.; Shao, Q.; Gao, Y.Q.; Raushel, F.M.
    A combined theoretical and experimental study of the ammonia tunnel in carbamoyl phosphate synthetase (2009), J. Am. Chem. Soc., 131, 10211-10219.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 6.3.5.5

Cloned(Commentary)

Cloned (Comment) Organism
expression of mutant enzymes in Escherichia coli strain RC50 Escherichia coli

Protein Variants

Protein Variants Comment Organism
A251C site-directed mutagenesis in the ammonia tunnel, analysis of secondary structure by circular dichroism measurements Escherichia coli
A311L site-directed mutagenesis in the ammonia tunnel, analysis of secondary structure by circular dichroism measurements Escherichia coli
A314C site-directed mutagenesis in the ammonia tunnel, analysis of secondary structure by circular dichroism measurements Escherichia coli
C232G/A251G/A314G site-directed mutagenesis in the ammonia tunnel, analysis of secondary structure by circular dichroism measurements Escherichia coli
C232V/A251V/A314V site-directed mutagenesis in the ammonia tunnel, analysis of secondary structure by circular dichroism measurements Escherichia coli
I352F site-directed mutagenesis in the ammonia tunnel, analysis of secondary structure by circular dichroism measurements Escherichia coli
S35Y site-directed mutagenesis in the ammonia tunnel, analysis of secondary structure by circular dichroism measurements Escherichia coli
T249V site-directed mutagenesis in the ammonia tunnel, analysis of secondary structure by circular dichroism measurements Escherichia coli

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+
-
 Escherichia coli

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
42000
-
 1 * 42000, small subunit, + 118000, large subunit Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2 ATP + L-glutamine + HCO3- + H2O Escherichia coli
-
 2 ADP + phosphate + L-glutamate + carbamoyl phosphate carbamoyl phosphate is utilized in the biosynthesis of arginine and pyrimidine nucleotides ?

Organism

Organism UniProt Comment Textmining
Escherichia coli
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant mutant enzymes from Escherichia coli strain RC50 to over 95% purity Escherichia coli

Reaction

Reaction Comment Organism Reaction ID
2 ATP + L-glutamine + hydrogencarbonate + H2O = 2 ADP + phosphate + L-glutamate + carbamoyl phosphate reaction mechanism with four separate reactions and three discrete, reactive, and unstable intermediates involved, overview. The small subunit contains the active site for the hydrolysis of glutamine, which is hydrolyzed to glutamate and ammonia via a thioester intermediate. The ammonia derived from this reaction travels 45 A to the active site for the synthesis of the carboxy phosphate intermediate, located in the N-terminal half of the large subunit. The carbamate intermediate, formed by the reaction of ammonia with carboxy phosphate, is subsequently channeled 45 A to the active site, located at the C-terminus of the large subunit, for the synthesis of carbamoyl phosphate, which is phosphorylated by the second molecule of MgATP to form the ultimate product carbamoyl phosphate. Molecular dynamics simulations, kinetic and dynamic of the mechanism for ammonia migration at the atomic level, free-energy profile for the remainder of the ammonia tunnel within CPS using the umbrella sampling technique, overview Escherichia coli
a

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2 ATP + L-glutamine + HCO3- + H2O
-
 Escherichia coli 2 ADP + phosphate + L-glutamate + carbamoyl phosphate
-
 ?
2 ATP + L-glutamine + HCO3- + H2O
-
 Escherichia coli 2 ADP + phosphate + L-glutamate + carbamoyl phosphate carbamoyl phosphate is utilized in the biosynthesis of arginine and pyrimidine nucleotides ?

Subunits

Subunits Comment Organism
heterodimer 1 * 42000, small subunit, + 118000, large subunit Escherichia coli
More presence of three spatially distinct active sites that are linked by two long molecular tunnels that extend approximately 100 A from one end of the protein to the other Escherichia coli

Synonyms

Synonyms Comment Organism
carbamoyl phosphate synthetase
-
 Escherichia coli
CPS
-
 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