Cloned (Comment) | Organism |
---|---|
expression of mutant enzymes in Escherichia coli strain RC50 | Escherichia coli |
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 | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | - |
Escherichia coli |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
42000 | - |
1 * 42000, small subunit, + 118000, large subunit | Escherichia coli |
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 | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant mutant enzymes from Escherichia coli strain RC50 to over 95% purity | Escherichia coli |
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 |
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 | 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 | Comment | Organism |
---|---|---|
carbamoyl phosphate synthetase | - |
Escherichia coli |
CPS | - |
Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.6 | - |
assay at | Escherichia coli |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Escherichia coli |