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Information on EC 6.3.5.5 - carbamoyl-phosphate synthase (glutamine-hydrolysing)
for references in articles please use BRENDA:EC6.3.5.5
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EC Tree 6 Ligases
6.3 Forming carbon-nitrogen bonds
6.3.5 Carbon-nitrogen ligases with glutamine as amido-N-donor
6.3.5.5 carbamoyl-phosphate synthase (glutamine-hydrolysing)
IUBMB Comments
The product carbamoyl phosphate is an intermediate in the biosynthesis of arginine and the pyrimidine nucleotides . The enzyme from Escherichia coli has three separate active sites, which are connected by a molecular tunnel that is almost 100 Å in length . The amidotransferase domain within the small subunit of the enzyme hydrolyses glutamine to ammonia via a thioester intermediate. The ammonia migrates through the interior of the protein, where it reacts with carboxyphosphate to produce the carbamate intermediate. The carboxyphosphate intermediate is formed by the phosphorylation of hydrogencarbonate by ATP at a site contained within the N-terminal half of the large subunit. The carbamate intermediate is transported through the interior of the protein to a second site within the C-terminal half of the large subunit, where it is phosphorylated by another ATP to yield the final product, carbamoyl phosphate . cf. EC 6.3.4.16, carbamoyl-phosphate synthase (ammonia).
The enzyme appears in viruses and cellular organisms
- 6.3.5.5
- pyrimidine
- dihydroorotase
- cpsases
-
transcarbamoylase
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2.1.3.2
-
ornithine-urea
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2.7.2.9
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3.5.2.3
- acivicin
-
caspase-activated
- dhoase
-
ureotelic
- atcase
- lungfish
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ammonia-dependent
-
carab
-
5-phosphoribosyl
- 1-pyrophosphate
- protopterus
- n-acetyl-l-glutamate
- drug development
- medicine
Reaction Schemes
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Synonyms
cps iii, cad protein, carbamoyl phosphate synthetase iii, carbamoyl phosphate synthetase ii, cpsii, carbamoyl-phosphate synthetase 2, glutamine-dependent carbamyl phosphate synthetase, carbamoyl phosphate synthetase (glutamine-hydrolyzing), cpsase type ii, carbamylphosphate synthetase - aspartate transcarbamylase, more
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SYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
CAD
Carbamoyl phosphate synthase (glutamine)
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-
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carbamoyl phosphate synthetase
Carbamoyl phosphate synthetase (glutamine-hydrolyzing)
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carbamoyl phosphate synthetase III
carbamoyl-phosphate synthetase
Carbamoyl-phosphate synthetase (glutamine-hydrolysing)
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Carbamoylphosphate synthase
Carbamoylphosphate synthetase
Carbamoylphosphate synthetase II
Carbamyl phosphate synthetase (glutamine)
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Carbamyl phosphate sythetase II
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carbamylphosphate synthetase - aspartate transcarbamylase
multifunctional protein
CPA2
CPS
CPS III
CPS1
CPSase
CPSase-A
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-
-
-
CPSase-P
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-
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EC 2.7.2.9
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formerly
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Glutamine-dependent carbamyl phosphate synthetase
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MGG_04503
Synthase, carbamoylphosphate (glutamine)
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Synthetase, carbamoylphosphate (glutamine-hydrolyzing)
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additional information
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carbamoyl phosphate synthetase is a member of the amidotransferase family of enzymes
Carbamoylphosphate synthase
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Carbamoylphosphate synthetase
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Carbamoylphosphate synthetase II
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CPS
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-
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CPSase
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REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
2 ATP + L-glutamine + hydrogencarbonate + H2O = 2 ADP + phosphate + L-glutamate + carbamoyl phosphate
ATP + carbamate = ADP + carbamoyl phosphate
(1d)
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-
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ATP + hydrogencarbonate = ADP + carboxyphosphate
(1b)
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L-glutamine + H2O = L-glutamate + NH3
(1a)
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-
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NH3 + carboxyphosphate = carbamate + phosphate
(1c)
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-
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2 ATP + L-glutamine + hydrogencarbonate + H2O = 2 ADP + phosphate + L-glutamate + carbamoyl phosphate
mechanism, formation of enzyme-bound carboxy phosphate
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2 ATP + L-glutamine + hydrogencarbonate + H2O = 2 ADP + phosphate + L-glutamate + carbamoyl phosphate
partially random sequential mechanism in which the ordered addition of MgATP2- , HCO3-, and Gln is followed by the ordered release of Glu and phosphate. The formation of carbamate from MgATP2-, HCO3-, and Gln proceeds via a stepwise, not concerted mechanism, involving at least one kinetically competent covalent intermediate, such as carboxyphosphate
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2 ATP + L-glutamine + hydrogencarbonate + H2O = 2 ADP + phosphate + L-glutamate + carbamoyl phosphate
mechanism involves at least three steps, the first of which is an ATP-dependent formation of enzyme-bound activated carbon dioxide
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2 ATP + L-glutamine + hydrogencarbonate + H2O = 2 ADP + phosphate + L-glutamate + carbamoyl phosphate
mechanism
-
2 ATP + L-glutamine + hydrogencarbonate + H2O = 2 ADP + phosphate + L-glutamate + carbamoyl phosphate
two residues, Ser948 and Thr1042, appear crucial for allosteric regulation of enzyme
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2 ATP + L-glutamine + hydrogencarbonate + H2O = 2 ADP + phosphate + L-glutamate + carbamoyl phosphate
effects of pressure on the partial reactions, study of the function of monomers of enzyme domains under conditions of pressure-induced dissociation
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2 ATP + L-glutamine + hydrogencarbonate + H2O = 2 ADP + phosphate + L-glutamate + carbamoyl phosphate
mechanism
-
2 ATP + L-glutamine + hydrogencarbonate + H2O = 2 ADP + phosphate + L-glutamate + carbamoyl phosphate
catalytic mechanism
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2 ATP + L-glutamine + hydrogencarbonate + H2O = 2 ADP + phosphate + L-glutamate + carbamoyl phosphate
An ordered uni bi mechanism for glutamine hydrolysis that is consistent with the isotope effects and with the catalytic properties of the enzyme is proposed
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2 ATP + L-glutamine + hydrogencarbonate + H2O = 2 ADP + phosphate + L-glutamate + carbamoyl phosphate
four partial reaction occurs at three active sites connected by a molecular tunnel of 100 Angstrom, reaction mechanism
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2 ATP + L-glutamine + hydrogencarbonate + H2O = 2 ADP + phosphate + L-glutamate + carbamoyl phosphate
the mammalian enzyme is part of carbamoyl-phosphate synthase-aspartate carbamoyltransferase-dihydroorotase, CAD, the carbamoyl phosphate synthesis requires the concerted action of the glutaminase and carbamoyl-phosphate synthetase domains of CAD
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2 ATP + L-glutamine + hydrogencarbonate + H2O = 2 ADP + phosphate + L-glutamate + carbamoyl phosphate
steps of reaction pathway for glutamine hydrolysis, the Michalis complex, the tetrahedral intermediate and glutamyl-thioester intermediate models, stereochemistry of the overall glutaminase reaction
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2 ATP + L-glutamine + hydrogencarbonate + H2O = 2 ADP + phosphate + L-glutamate + carbamoyl phosphate
pathway
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2 ATP + L-glutamine + hydrogencarbonate + H2O = 2 ADP + phosphate + L-glutamate + carbamoyl phosphate
reaction pathway of the enzyme small subunit proceeds through the formation of the glutamyl thioester intermediate
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2 ATP + L-glutamine + hydrogencarbonate + H2O = 2 ADP + phosphate + L-glutamate + carbamoyl phosphate
in glutamine utilizing enzymes, the hydrolysis of glutamine to yield ammonia is catalyzed at a triad-type glutamine amidotransferase domain
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2 ATP + L-glutamine + hydrogencarbonate + H2O = 2 ADP + phosphate + L-glutamate + carbamoyl phosphate
pathway, relationship between mechanism and structure
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2 ATP + L-glutamine + hydrogencarbonate + H2O = 2 ADP + phosphate + L-glutamate + carbamoyl phosphate
the chemical mechanism of the bicarbonate-dependent ATPase reaction of enzyme involves carboxy phosphate, but not CO2, as an reactive intermediate
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2 ATP + L-glutamine + hydrogencarbonate + H2O = 2 ADP + phosphate + L-glutamate + carbamoyl phosphate
reaction mechanism, the ammonium is channeled as ammonia through a channel formed by the small subunit by forming and breaking hydrogen bonds to Gly292, Ser35, Pro358, Gly293, and Thr37 in a stepwise fashion as it travels through the hydrophilic passage toward the subunit interface
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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
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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
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2 ATP + L-glutamine + hydrogencarbonate + H2O = 2 ADP + phosphate + L-glutamate + carbamoyl phosphate
carbamate, the product of the reaction involving ATP, bicarbonate, and ammonia, must be delivered from the site of formation to the site of utilization by traveling nearly 40 A within the enzyme. The tunnel is composed of three continuous water pockets and two narrow connecting parts, near residues A23 and G575. The two narrow parts render two free energy barriers of 6.7 and 8.4 kcal/mol, respectively. Three water pockets are filled with about 21, 9, and 9 waters, respectively, and the corresponding relative free energies of carbamate residing in these free energy minima are 5.8, 0, and 1.6 kcal/mol, respectively. The release of phosphate into solution at the site for the formation of carbamate allows the side chain of R306 to rotate toward E25, E383, and E604. This rotation is virtually prohibited by a barrier of at least 23 kcal/mol when phosphate remains bound. This conformational change not only opens the entrance of the tunnel but also shields the charge-charge repulsion from the three glutamate residues when carbamate passes through the tunnel
2 ATP + L-glutamine + hydrogencarbonate + H2O = 2 ADP + phosphate + L-glutamate + carbamoyl phosphate
mechanism involves at least three steps, the first of which is an ATP-dependent formation of enzyme-bound activated carbon dioxide
Escherichia coli B / ATCC 11303
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-
2 ATP + L-glutamine + hydrogencarbonate + H2O = 2 ADP + phosphate + L-glutamate + carbamoyl phosphate
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
amide group transfer
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amination
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Phosphorylation
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PATHWAY SOURCE
PATHWAYS
BRENDA
MetaCyc
L-arginine biosynthesis I (via L-ornithine), L-arginine biosynthesis II (acetyl cycle), L-arginine biosynthesis III (via N-acetyl-L-citrulline), L-arginine biosynthesis IV (archaea), UMP biosynthesis I, UMP biosynthesis II, UMP biosynthesis III
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SYSTEMATIC NAME
IUBMB Comments
hydrogen-carbonate:L-glutamine amido-ligase (ADP-forming, carbamate-phosphorylating)
The product carbamoyl phosphate is an intermediate in the biosynthesis of arginine and the pyrimidine nucleotides [4]. The enzyme from Escherichia coli has three separate active sites, which are connected by a molecular tunnel that is almost 100 A in length [8]. The amidotransferase domain within the small subunit of the enzyme hydrolyses glutamine to ammonia via a thioester intermediate. The ammonia migrates through the interior of the protein, where it reacts with carboxyphosphate to produce the carbamate intermediate. The carboxyphosphate intermediate is formed by the phosphorylation of hydrogencarbonate by ATP at a site contained within the N-terminal half of the large subunit. The carbamate intermediate is transported through the interior of the protein to a second site within the C-terminal half of the large subunit, where it is phosphorylated by another ATP to yield the final product, carbamoyl phosphate [6]. cf. EC 6.3.4.16, carbamoyl-phosphate synthase (ammonia).
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CAS REGISTRY NUMBER
COMMENTARY
37233-48-0
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SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
LITERATURE
COMMENTARY
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
2 ATP + hydrazine + HCO3-
2 ADP + phosphate + N-amino carbamoyl phosphate
-
Substrates: -
Products: -
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?
2 ATP + hydroxylamine + HCO3-
2 ADP + phosphate + N-hydroxy carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-glutamine + HCO3- + H2O + H+
2 ADP + phosphate + L-glutamate + carbamoyl phosphate
ATP + L-glutamine + HCO3- + H2O
2 ADP + phosphate + L-glutamate + carbamoyl phosphate
-
Substrates: there is no preferential partitioning of carbamoyl phosphate between the arginine and pyrimidine biosynthetic pathways. Channeling must occur during the dynamic association of coupled enzymes pairs. The interaction of carbamoyl-phosphate synthetase/aspartate transcarbamoylase is demonstrated by the unexpectedly weak inhibition of the coupled reaction by the bisubstrate analog, N-(phosphonacetyl)-L-aspartate. In the coupled reaction, the effective concentration of carbamoyl phosphate in the vicinity of the aspartate transcarbamoylase active site is 96fold higher than the concentration in the bulk phase. Channeling probably plays an essential role in protecting this very unstable intermediate of metabolic pathways performing at extreme temperatures
Products: -
Products: -
?
ATP + L-glutamine + hydrogencarbonate + H2O
ADP + phosphate + L-glutamate + carbamoyl phosphate
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: the product carbamoyl phosphate is the initial intermediate in the biosynthesis of both pyrimidine and arginine
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: enzyme is a key enzyme in the pyrimidine nucleotide and arginine biosynthetic pathways
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
Substrates: -
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Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
ir
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
649457, 649849, 649877, 649883, 649886, 649895, 649964, 649986, 650020, 650110, 650289, 650798, 650936, 651292, 651471, 651882, 651901, 651952, 652028, 652201, 652334, 652351, 652418, 652788, 652823
Substrates: -
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: overall reaction is irreversible
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: enzyme is a key enzyme in the pyrimidine nucleotide and arginine biosynthetic pathways
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: the enzyme catalyses the entry and rate-limiting step of the urea cycle
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: the enzyme catalyses the first step in the synthesis of arginine and pyrimidine
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: the product carbamoyl phosphate is an essential precursor of arginine and pyrimidine biosynthesis
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: the product carbamoyl phosphate is utilized for the arginine biosynthesis
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: the product carbamoyl phosphate plays a critical role in both arginine and pyrimidine biosynthesis
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: the product carbamoylphosphate is utilized in the synthesis of arginine and pyrimidine nucleotides and in the urea cycle
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: carbamoyl phosphate synthetase contains an internal molecular tunnel, which has been proposed to facilitate the translocation of reaction intermediates from one active site to another. Ammonia, the product from the hydrolysis of glutamine in the small subunit, is apparently transported to the next active site in the large subunit of CPS over a distance of about 45 Å. The ammonia tunnel that connects these two active sites provides a direct path for the guided diffusion of ammonia and protection from protonation
Products: -
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?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
Escherichia coli B / ATCC 11303
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: enzyme is a key enzyme in the pyrimidine nucleotide and arginine biosynthetic pathways
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
Fresh-water teleost
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
Substrates: -
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
Substrates: the enzyme catalyses the first step in the arginine and pyrimidine biosynthesis
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
Substrates: -
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
Substrates: enzyme is a key enzyme in the pyrimidine nucleotide and arginine biosynthetic pathways, possible role in the pathway for the synthesis of exopolysaccharides
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: enzyme catalyses the first step of urea cycle in fish
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
Substrates: -
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: cross-talk between domain N and C is required for product release from subunit C
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: the product carbamoyl phosphate is utilized for the arginine biosynthesis
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: the product carbamoyl phosphate is utilized for the arginine biosynthesis
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
Substrates: -
Products: -
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?
2 ATP + L-Gln + HCO3- + H+ + H2O
2 ADP + phosphate + L-glutamate + carbamoyl phosphate
-
Substrates: the product carbamoyl phosphate is utilized in the pyrimidine and arginine biosynthetic pathways
Products: -
Products: -
?
2 ATP + L-Gln + HCO3- + H+ + H2O
2 ADP + phosphate + L-glutamate + carbamoyl phosphate
Substrates: -
Products: -
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?
2 ATP + L-glutamine + HCO3- + H2O
2 ADP + phosphate + L-glutamate + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-glutamine + HCO3- + H2O
2 ADP + phosphate + L-glutamate + carbamoyl phosphate
-
Substrates: -
Products: carbamoyl phosphate is utilized in the biosynthesis of arginine and pyrimidine nucleotides
Products: carbamoyl phosphate is utilized in the biosynthesis of arginine and pyrimidine nucleotides
?
2 ATP + L-glutamine + HCO3- + H2O
2 ADP + phosphate + L-glutamate + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-glutamine + HCO3- + H2O
2 ADP + phosphate + L-glutamate + carbamoyl phosphate
Substrates: -
Products: -
Products: -
?
2 ATP + L-glutamine + HCO3- + H2O + H+
2 ADP + phosphate + L-glutamate + carbamoyl phosphate
-
Substrates: a single carbamoyl phosphate synthetase supplies both the pyrimidine and arginine pathways of this organism
Products: -
Products: -
?
2 ATP + L-glutamine + HCO3- + H2O + H+
2 ADP + phosphate + L-glutamate + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-glutamine + HCO3- + H2O + H+
2 ADP + phosphate + L-glutamate + carbamoyl phosphate
Sulfolobus acidocaldarius DG6 / ATCC 49426
-
Substrates: a single carbamoyl phosphate synthetase supplies both the pyrimidine and arginine pathways of this organism
Products: -
Products: -
?
2 ATP + L-glutamine + HCO3- + H2O + H+
2 ADP + phosphate + L-glutamate + carbamoyl phosphate
Sulfolobus acidocaldarius DG6 / ATCC 49426
-
Substrates: -
Products: -
Products: -
?
2 ATP + NH4+ + HCO3-
2 ADP + phosphate + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + NH4+ + HCO3-
2 ADP + phosphate + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + NH4+ + HCO3-
2 ADP + phosphate + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + NH4+ + HCO3-
2 ADP + phosphate + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + NH4+ + HCO3-
2 ADP + phosphate + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
ir
2 ATP + NH4+ + HCO3-
2 ADP + phosphate + carbamoyl phosphate
-
649849, 649877, 649883, 649886, 649895, 649964, 650936, 651471, 651901, 651952, 652028, 652334, 652418, 652823, 1744
Substrates: -
Products: -
Products: -
?
2 ATP + NH4+ + HCO3-
2 ADP + phosphate + carbamoyl phosphate
-
Substrates: the enzyme is able to utilize external NH4+ as an alternative nitrogen source when glutamine is absent
Products: -
Products: -
?
2 ATP + NH4+ + HCO3-
2 ADP + phosphate + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + NH4+ + HCO3-
2 ADP + phosphate + carbamoyl phosphate
Escherichia coli B / ATCC 11303
-
Substrates: -
Products: -
Products: -
?
2 ATP + NH4+ + HCO3-
2 ADP + phosphate + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + NH4+ + HCO3-
2 ADP + phosphate + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + NH4+ + HCO3-
2 ADP + phosphate + carbamoyl phosphate
Substrates: -
Products: -
Products: -
?
2 ATP + NH4+ + HCO3-
2 ADP + phosphate + carbamoyl phosphate
-
Substrates: CPS III shows low activity in comparison to L-glutamine
Products: -
Products: -
?
2 ATP + NH4+ + HCO3-
2 ADP + phosphate + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + NH4+ + HCO3-
2 ADP + phosphate + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + NH4+ + HCO3-
2 ADP + phosphate + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + NH4+ + HCO3-
2 ADP + phosphate + carbamoyl phosphate
-
Substrates: utilizes NH4+ with 600times lower affinity than Gln
Products: -
Products: -
?
2 ATP + NH4+ + HCO3-
2 ADP + phosphate + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + NH4+ + HCO3-
2 ADP + phosphate + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + NH4+ + HCO3-
2 ADP + phosphate + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + NH4+ + HCO3-
2 ADP + phosphate + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + NH4+ + HCO3-
2 ADP + phosphate + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + NH4+ + HCO3-
2 ADP + phosphate + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
ATP + L-Gln + HCO3-
?
-
Substrates: catalyzes the first step of de novo pyrimidine biosynthesis
Products: -
Products: -
?
ATP + L-Gln + HCO3-
?
-
Substrates: functions in the synthesis of urea used in osmoregulation
Products: -
Products: -
?
ATP + L-Gln + HCO3-
?
-
Substrates: catalyzes the first step of de novo pyrimidine biosynthesis
Products: -
Products: -
?
ATP + L-Gln + HCO3-
?
-
Substrates: important role of insulin in regulation of liver carbamoyl-phosphate synthase II and lack of responsiveness of the hepatoma enzyme to insulin concentration
Products: -
Products: -
?
ATP + L-Gln + HCO3-
?
-
Substrates: catalyzes the first step of de novo pyrimidine biosynthesis
Products: -
Products: -
?
ATP + L-Gln + HCO3-
?
-
Substrates: catalyzes the synthesis of all carbamoyl phosphate which serves two metabolic functions: it is an intermediate in synthesis of arginine and in the synthesis of pyrimidines
Products: -
Products: -
?
ATP + L-glutamine + HCO3- + H2O
ADP + phosphate + L-glutamate + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
ATP + L-glutamine + HCO3- + H2O
ADP + phosphate + L-glutamate + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
ATP + L-glutamine + HCO3- + H2O
ADP + phosphate + L-glutamate + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
ATP + L-glutamine + HCO3- + H2O
ADP + phosphate + L-glutamate + carbamoyl phosphate
-
Substrates: substrate-channelling, the enzyme is also active with NH3 as nitrogen source, cf. EC 6.3.4.16
Products: -
Products: -
?
ATP + L-glutamine + HCO3- + H2O
ADP + phosphate + L-glutamate + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
ATP + L-glutamine + HCO3- + H2O
ADP + phosphate + L-glutamate + carbamoyl phosphate
-
Substrates: substrate-channelling, the enzyme is also active with NH3 as nitrogen source, cf. EC 6.3.4.16
Products: -
Products: -
?
ATP + L-glutamine + HCO3- + H2O
ADP + phosphate + L-glutamate + carbamoyl phosphate
-
Substrates: CPS1 plays a critical role in the biosynthesis of pyrimidine nucleotides and the detoxification of ammonia
Products: -
Products: -
?
ATP + L-glutamine + HCO3- + H2O
ADP + phosphate + L-glutamate + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
ATP + L-glutamine + HCO3- + H2O
ADP + phosphate + L-glutamate + carbamoyl phosphate
-
Substrates: the enzyme from Escherichia coli catalyzes the synthesis of carbamoyl phosphate from bicarbonate, glutamine, and two molecules of ATP via a chemical mechanism that involves four separate reactions and three unstable intermediates, e.g. carboxyphosphate and carbamate
Products: -
Products: -
r
ATP + L-glutamine + HCO3- + H2O
ADP + phosphate + L-glutamate + carbamoyl phosphate
-
Substrates: neither residue P909 nor residue G919 is critical for eCPS function due to the absence of vicinal cysteinyl residues in wild-type eCPS
Products: -
Products: -
?
ATP + L-glutamine + HCO3- + H2O
ADP + phosphate + L-glutamate + carbamoyl phosphate
-
Substrates: 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
Products: -
Products: -
?
ATP + L-glutamine + HCO3- + H2O
ADP + phosphate + L-glutamate + carbamoyl phosphate
-
Substrates: allosteric regulation and CPSII regulatory domains, overview
Products: -
Products: -
?
ATP + L-glutamine + HCO3- + H2O
ADP + phosphate + L-glutamate + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
ATP + L-glutamine + hydrogencarbonate + H2O
ADP + phosphate + L-glutamate + carbamoyl phosphate
-
Substrates: ATP in form of MgATP2-
Products: -
Products: -
?
ATP + L-glutamine + hydrogencarbonate + H2O
ADP + phosphate + L-glutamate + carbamoyl phosphate
-
Substrates: ATP in form of MgATP2-
Products: -
Products: -
?
additional information
?
-
-
Substrates: assay method development based on the fixation of 14C from the substrate bicarbonate to give radioactive products. In the coupled assay the carbamoyl phosphate product of the enzyme is converted to carbamoyl aspartate in the presence of aspartate and aspartate transcarbamoylase, substrate-channelling between aspartate aminotransferase and CPSase, overview
Products: -
Products: -
?
additional information
?
-
-
Substrates: assay method development based on the fixation of 14C from the substrate bicarbonate to give radioactive products. In the coupled assay the carbamoyl phosphate product of the enzyme is converted to carbamoyl aspartate in the presence of aspartate and aspartate transcarbamoylase, substrate-channelling between aspartate aminotransferase and CPSase, overview
Products: -
Products: -
?
additional information
?
-
-
Substrates: existence of an alternate route to the bicarbonate phosphorylation site when ammonia is provided as an external nitrogen source
Products: -
Products: -
?
additional information
?
-
-
Substrates: CPS uses the hydrolysis of glutamine as a localized source of ammonia for biosynthetic transformations
Products: -
Products: -
?
additional information
?
-
-
Substrates: the smaller subunit contains the binding site for the hydrolysis of glutamine, whereas the large subunit catalyzes the formation of carbamoyl phosphate. Molecular dynamics simulations and free-energy profile for the transfer of ammonia and ammonium through a tunnel in the small subunit of CPS, by use of the enzyme's crystal structure, resulting in five successful trajectories for ammonia transfer, while ammonium is immobilized in a water pocket inside the small subunit of the heterodimeric protein, the carbamate tunnel connects the two active sites within the large subunit to one another and guides the unstable carbamate intermediate to the site of phosphorylation, mechanism, overview
Products: -
Products: -
?
additional information
?
-
-
Substrates: the enzyme is organized as CAD, i.e. carbamoyl-phosphate synthetase 2, aspartate transcarbamylase, and dihydroorotase, a tripartite enzyme that catalyzes the first three steps of pyrimidine biosynthesis
Products: -
Products: -
?
additional information
?
-
-
Substrates: hydrolysis of L-Gln in absence of the other substrates, at 5.9% the rate of glutamine hydrolysis in the complete system
Products: -
Products: -
?
additional information
?
-
-
Substrates: the gene for carbamoylphosphate synthetase (carAB) is induced by FlhD and FlhC
Products: -
Products: -
?
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
LITERATURE
COMMENTARY
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
2 ATP + L-Gln + HCO3- + H+ + H2O
2 ADP + phosphate + L-glutamate + carbamoyl phosphate
-
Substrates: the product carbamoyl phosphate is utilized in the pyrimidine and arginine biosynthetic pathways
Products: -
Products: -
?
2 ATP + L-glutamine + HCO3- + H2O + H+
2 ADP + phosphate + L-glutamate + carbamoyl phosphate
ATP + L-glutamine + HCO3- + H2O
2 ADP + phosphate + L-glutamate + carbamoyl phosphate
-
Substrates: there is no preferential partitioning of carbamoyl phosphate between the arginine and pyrimidine biosynthetic pathways. Channeling must occur during the dynamic association of coupled enzymes pairs. The interaction of carbamoyl-phosphate synthetase/aspartate transcarbamoylase is demonstrated by the unexpectedly weak inhibition of the coupled reaction by the bisubstrate analog, N-(phosphonacetyl)-L-aspartate. In the coupled reaction, the effective concentration of carbamoyl phosphate in the vicinity of the aspartate transcarbamoylase active site is 96fold higher than the concentration in the bulk phase. Channeling probably plays an essential role in protecting this very unstable intermediate of metabolic pathways performing at extreme temperatures
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: the product carbamoyl phosphate is the initial intermediate in the biosynthesis of both pyrimidine and arginine
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: enzyme is a key enzyme in the pyrimidine nucleotide and arginine biosynthetic pathways
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
ir
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: enzyme is a key enzyme in the pyrimidine nucleotide and arginine biosynthetic pathways
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: the enzyme catalyses the entry and rate-limiting step of the urea cycle
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: the enzyme catalyses the first step in the synthesis of arginine and pyrimidine
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: the product carbamoyl phosphate is an essential precursor of arginine and pyrimidine biosynthesis
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: the product carbamoyl phosphate is utilized for the arginine biosynthesis
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: the product carbamoyl phosphate plays a critical role in both arginine and pyrimidine biosynthesis
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: the product carbamoylphosphate is utilized in the synthesis of arginine and pyrimidine nucleotides and in the urea cycle
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: enzyme is a key enzyme in the pyrimidine nucleotide and arginine biosynthetic pathways
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
Substrates: the enzyme catalyses the first step in the arginine and pyrimidine biosynthesis
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
Substrates: enzyme is a key enzyme in the pyrimidine nucleotide and arginine biosynthetic pathways, possible role in the pathway for the synthesis of exopolysaccharides
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: enzyme catalyses the first step of urea cycle in fish
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: the product carbamoyl phosphate is utilized for the arginine biosynthesis
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: the product carbamoyl phosphate is utilized for the arginine biosynthesis
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-Gln + HCO3-
2 ADP + phosphate + L-Glu + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-glutamine + HCO3- + H2O
2 ADP + phosphate + L-glutamate + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
2 ATP + L-glutamine + HCO3- + H2O
2 ADP + phosphate + L-glutamate + carbamoyl phosphate
-
Substrates: -
Products: carbamoyl phosphate is utilized in the biosynthesis of arginine and pyrimidine nucleotides
Products: carbamoyl phosphate is utilized in the biosynthesis of arginine and pyrimidine nucleotides
?
2 ATP + L-glutamine + HCO3- + H2O
2 ADP + phosphate + L-glutamate + carbamoyl phosphate
Substrates: -
Products: -
Products: -
?
2 ATP + L-glutamine + HCO3- + H2O + H+
2 ADP + phosphate + L-glutamate + carbamoyl phosphate
-
Substrates: a single carbamoyl phosphate synthetase supplies both the pyrimidine and arginine pathways of this organism
Products: -
Products: -
?
2 ATP + L-glutamine + HCO3- + H2O + H+
2 ADP + phosphate + L-glutamate + carbamoyl phosphate
Sulfolobus acidocaldarius DG6 / ATCC 49426
-
Substrates: a single carbamoyl phosphate synthetase supplies both the pyrimidine and arginine pathways of this organism
Products: -
Products: -
?
ATP + L-Gln + HCO3-
?
-
Substrates: catalyzes the first step of de novo pyrimidine biosynthesis
Products: -
Products: -
?
ATP + L-Gln + HCO3-
?
-
Substrates: functions in the synthesis of urea used in osmoregulation
Products: -
Products: -
?
ATP + L-Gln + HCO3-
?
-
Substrates: catalyzes the first step of de novo pyrimidine biosynthesis
Products: -
Products: -
?
ATP + L-Gln + HCO3-
?
-
Substrates: important role of insulin in regulation of liver carbamoyl-phosphate synthase II and lack of responsiveness of the hepatoma enzyme to insulin concentration
Products: -
Products: -
?
ATP + L-Gln + HCO3-
?
-
Substrates: catalyzes the first step of de novo pyrimidine biosynthesis
Products: -
Products: -
?
ATP + L-Gln + HCO3-
?
-
Substrates: catalyzes the synthesis of all carbamoyl phosphate which serves two metabolic functions: it is an intermediate in synthesis of arginine and in the synthesis of pyrimidines
Products: -
Products: -
?
ATP + L-glutamine + HCO3- + H2O
ADP + phosphate + L-glutamate + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
ATP + L-glutamine + HCO3- + H2O
ADP + phosphate + L-glutamate + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
ATP + L-glutamine + HCO3- + H2O
ADP + phosphate + L-glutamate + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
ATP + L-glutamine + HCO3- + H2O
ADP + phosphate + L-glutamate + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
ATP + L-glutamine + HCO3- + H2O
ADP + phosphate + L-glutamate + carbamoyl phosphate
-
Substrates: CPS1 plays a critical role in the biosynthesis of pyrimidine nucleotides and the detoxification of ammonia
Products: -
Products: -
?
ATP + L-glutamine + HCO3- + H2O
ADP + phosphate + L-glutamate + carbamoyl phosphate
-
Substrates: -
Products: -
Products: -
?
ATP + L-glutamine + HCO3- + H2O
ADP + phosphate + L-glutamate + carbamoyl phosphate
-
Substrates: allosteric regulation and CPSII regulatory domains, overview
Products: -
Products: -
?
additional information
?
-
-
Substrates: CPS uses the hydrolysis of glutamine as a localized source of ammonia for biosynthetic transformations
Products: -
Products: -
?
additional information
?
-
-
Substrates: the enzyme is organized as CAD, i.e. carbamoyl-phosphate synthetase 2, aspartate transcarbamylase, and dihydroorotase, a tripartite enzyme that catalyzes the first three steps of pyrimidine biosynthesis
Products: -
Products: -
?
additional information
?
-
-
Substrates: the gene for carbamoylphosphate synthetase (carAB) is induced by FlhD and FlhC
Products: -
Products: -
?
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
MgUTP2-
-
in presence of high concentrations of dimethyl sulfoxide, MgUTP2- stimulates at low concentrations, while it is inhibitory at higher concentrations
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
K+
Mg2+
Mn2+
-
maximal activity at concentration of Mn2+ approximately equal to the ATP concentration
NH4+
Mg2+
-
requires high concentrations of MgATP2- for optimal activity, half-maximal velocity at 10 mM MgATP2-
NH4+
-
positive allosteric effector of Gln-dependent activity, promotes self-association of the enzyme
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INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
acivicin
-
selective time-dependent inhibition of L-Gln-dependent activity, L-Gln protects the enzyme from inactivation. Stimulation of NH4+-dependent activity
Ala
-
inhibits glutamine hydrolysis and Gln-dependent carbamoyl phosphate synthesis
Alkyl hydrazines
-
inhibits Gln-dependent activity, but not NH4+-dependent activity
Arg
-
arginine-specific carbamoyl-phosphate synthase is repressed, pyrimidine-specific enzyme not
H2O2
-
0.2 mM, inhibits Gln-dependent activity. No effect on the activity with NH4+ in carbamoyl-phosphate synthase reaction
L-2-Amino-4-oxo-5-chloropentanoate
-
selective inactivation of Gln-dependent activity
MgUTP2-
-
in presence of high concentrations of dimethyl sulfoxide, MgUTP2- at low concentrations stimulates, while it is inhibitory at higher concentrations
N-(phosphonacetyl)-L-aspartate
-
there is no preferential partitioning of carbamoyl phosphate between the arginine and pyrimidine biosynthetic pathways. Channeling must occur during the dynamic association of coupled enzymes pairs. The interaction of carbamoyl-phosphate synthetase/aspartate transcarbamoylase is demonstrated by the unexpectedly weak inhibition of the coupled reaction by the bisubstrate analog, N-(phosphonacetyl)-L-aspartate
NEM
-
irreversible inactivation of synthetase activity. Increase of glutaminase activity
P1,P5-di(adenosine 5')-pentaphosphate
-
inhibits two partial reactions catalyzed by the enzyme: bicarbonate-dependent ATPase and ATP synthesis from carbamoyl phosphate
potassium cyanate
-
inhibits Gln-dependent activity, but not NH4+-dependent activity
hydroxylamine
-
inhibits Gln-dependent activity, but not NH4+-dependent activity; the inhibitory effect is much greater at pH 9 than at pH 6
hydroxylamine
-
inhibits Gln-dependent activity, but not NH4+-dependent activity
polyamines
-
inhibition is partially reversed by increasing the concentration of Mg2+ or MgATP2-, or by adding low concentrations of 5-phosphoribosyl 1-diphosphate
-
UMP
-
the IMP activator and the UMP inhibitor bind to the same site on the enzyme
UMP
-
product of the pyrimidine nucleotide pathway, acts as a negative allosteric effector for eCPS, which also catalyzes the first step of this pathway
UMP
-
carbamoyl-phosphate synthase belonging to the pyrimidine pathway, carbamoyl-phosphate synthase belonging to the arginine pathway is not inhibited
UMP
-
the nonresponsive enzyme can bind UMP at domain D and allosteric effect on chimeric enzyme
UTP
slight UTP inhibition of SU(b) mutant enzyme is probably due to the chelating of Mg2+ by UTP
UTP
-
when L-glutamine is the substrate: 94% inhibition of enzyme activity peak 1: CPS II, 6% inhibition of enzyme activity peak 2: CPS III, when NH4Cl is the substrate: 100% inhibition of enzyme activity peak 1: CPS II, 46% inhibition of enzyme activity peak 2: CPS III
UTP
competitive, feedback inhibitor, binds to a regulatory site located in the vicinity of the carbamylphosphate synthetase catalytic subsite
additional information
-
IMP binds to enzyme but the actual effect of this ligand is dependent upon temperature and assay conditions
-
additional information
-
IMP binds to enzyme but the actual effect of this ligand is dependent upon temperature and assay conditions
-
additional information
-
both the Luc-1b replicon and JFH1-2a virus expression inhibit CAD
-
additional information
-
acivicin inhibits the growth of Toxoplasma gondii
-
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
acivicin
-
selective time-dependent inhibition of L-Gln-dependent activity, L-Gln protects the enzyme from inactivation. Stimulation of NH4+-dependent activity
human Rad9 checkpoint protein
-
stimulates the carbamoyl phosphate synthetase activity of the multifunctional protein CAD. Rad9 binds to the Cpsase domain, and, moreover, this binding results in a 2fold stimulation of the CPSase activity of CAD
-
NEM
-
250fold activation of glutaminase activity. Irreversible inactivation of synthetase activity
phosphoribosyl 5'-diphosphate
-
activation of Escherichia coli and chimeric enzyme, the presence of UTP reduces the PRPP binding
5-phospho-alpha-D-ribosyl 1-diphosphate
-
pyrimidine-specific carbamoyl-phosphate synthase is activated, arginine-specific enzyme not
5-phospho-alpha-D-ribosyl 1-diphosphate
-
stimulates
IMP
-
the IMP activator and the UMP inhibitor bind to the same site on the enzyme
IMP
-
His995, located in the C-terminal regulatory domain, is cross-linked with IMP
IMP
-
allosteric effect on Escherichia coli enzyme in dependence of temperature, no response to chimeric enzyme
IMP
-
the nonresponsive enzyme can bind UMP at domain D and allosteric effect on chimeric enzyme
L-ornithine
-
binds to the enzymes large subunit at the interface located between the allosteric and the carbamoyl phosphate domains
L-ornithine
-
positive allosteric activation, E783, T1042, and T1043 are primarily responsible for the binding of ornithine to enzyme
L-ornithine
-
effector on ATP synthesis reaction, when ornithine is bound to the enzyme, its activation dominates the actions of UMP and IMP
L-ornithine
-
allosteric effect on chimeric enzyme and Escherichia coli enzyme
L-ornithine
-
allosteric effect on chimeric enzyme, no effect on Saccharomyces cerevisae enzyme
N-acetylglutamate
-
carbamoyl-phosphate synthase II: no requirement for N-acetylglutamate
N-acetylglutamate
-
carbamoyl-phosphate synthase II: no requirement for N-acetylglutamate
N-acetylglutamate
-
carbamoyl-phosphate synthase III: requirement for N-acetylglutamate
ornithine
-
co-substrate with carbamoyl phosphate for the second step of arginine biosynthesis, acts as a positive allosteric effector for eCPS, the catalyst for the first pathway step
phosphate
-
multifunctional proteins are phosphorylated in vivo and in vitro. This covalent modification has only a small effect on the enzymic activity, and might be a signal for protein degradation
phosphate
-
multifunctional proteins are phosphorylated in vivo and in vitro. This covalent modification has only a small effect on the enzymic activity, and might be a signal for protein degradation
additional information
-
activicin inhibits the Toxoplasma gondii growth
-
additional information
-
CPSII GATase domain as well as the C-terminal allosteric regulatory domain are essential, critical dependence on the apicomplexan CPSII GATase domain in vivo, overview
-
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DISEASE
TITLE OF PUBLICATION
LINK TO PUBMED
Carcinoma
Inactivation by acivicin of carbamoyl-phosphate synthetase II of human colon carcinoma.
Carcinoma
Initial steps in pyrimidine synthesis in Ehrlich ascites carcinoma in vitro. II. The synthesis of carbamyl phosphate by a soluble, glutamine-dependent carbamyl phosphate synthetase.
Carcinoma
RNAi knockdown of caspase-activated DNase inhibits rotenone-induced DNA fragmentation in HeLa cells.
Carcinoma, Hepatocellular
A multienzyme complex of carbamoyl-phosphate synthase (glutamine): aspartate carbamoyltransferase: dihydoorotase (rat ascites hepatoma cells and rat liver).
Carcinoma, Hepatocellular
Carbamoyl phosphate synthetase (glutamine-hydrolyzing): increased activity in cancer cells.
Carcinoma, Hepatocellular
In vivo inactivation by acivicin of carbamoyl-phosphate synthetase II in rat hepatoma.
Carcinoma, Hepatocellular
Increased carbamoyl-phosphate synthetase II concentration in rat hepatomas: immunological evidence.
Carcinoma, Hepatocellular
Increased messenger RNA concentration for carbamoyl-phosphate synthase II in hepatoma 3924A.
Carcinoma, Hepatocellular
Increased synthesis of carbamoyl-phosphate synthase II (EC 6.3.5.5) in hepatoma 3924A.
Carcinoma, Hepatocellular
Phosphorylation and dephosphorylation of carbamoyl-phosphate synthetase II complex of rat ascites hepatoma cells.
Carcinoma, Hepatocellular
Purification of homogeneous glutamine-dependent carbamyl phosphate synthetase from ascites hepatoma cells as a complex with aspartate transcarbamylase and dihydroorotase.
Carcinoma, Hepatocellular
Regulatory properties and behavior of activity of carbamoyl phosphate synthetase II (glutamine-hydrolyzing) in normal and proliferating tissues.
Chagas Disease
Molecular interaction of the first 3 enzymes of the de novo pyrimidine biosynthetic pathway of Trypanosoma cruzi.
Cholera
Effect of pre-existing immunity for systemic and mucosal immune responses to intranasal immunization with group B Streptococcus type III capsular polysaccharide-cholera toxin B subunit conjugate.
Colonic Neoplasms
Preparation and characterization of a novel monoclonal antibody specific to human CAD protein.
Colorectal Neoplasms
Antibody Responses to Streptococcus Gallolyticus Subspecies Gallolyticus Proteins in a Large Prospective Colorectal Cancer Cohort Consortium.
Infections
A Carbamoyl Phosphate Synthetase II (CPSII) Deletion Mutant of Toxoplasma gondii Induces Partial Protective Immunity in Mice.
Infections
Characterization of the type III capsular polysaccharide produced by Burkholderia pseudomallei.
Infections
Distinct Group B Streptococcus Sequence and Capsule Types Differentially Impact Macrophage Stress and Inflammatory Signaling Responses.
Neoplasms
Carbamoyl phosphate synthetase (glutamine-hydrolyzing): increased activity in cancer cells.
Neoplasms
Critical importance of the de novo pyrimidine biosynthesis pathway for Trypanosoma cruzi growth in the mammalian host cell cytoplasm.
Neoplasms
Targeting pyrimidine synthesis accentuates molecular therapy response in glioblastoma stem cells.
Neoplasms
[Histogenesis of gallbladder cancer with special reference to metaplastic changes and distribution of various mucins and CEA]
Sepsis
Recurrent sepsis and neuroinvasive disease in a neonate culture-positive for a Group B Streptococcus CPS III serotype, hvgA+ strain.
Starvation
Effects of phosphate limitation on expression of genes involved in pyrimidine synthesis and salvaging in Arabidopsis.
Stomach Neoplasms
Preparation and characterization of a novel monoclonal antibody specific to human CAD protein.
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.022
ADP
mutant A23K, ATP-synthesis, pH 7.6, temperature not specified in the publication
0.03
ADP
mutant G575F, ATP-synthesis, pH 7.6, temperature not specified in the publication
0.054
ADP
mutant G575K, ATP-synthesis, pH 7.6, temperature not specified in the publication
0.088
ADP
mutant M174E, ATP-synthesis, pH 7.6, temperature not specified in the publication
0.13
ADP
wild-type, ATP-synthesis, pH 7.6, temperature not specified in the publication
0.13
ADP
mutant M378E, ATP-synthesis, pH 7.6, temperature not specified in the publication
0.17
ADP
mutant M174E/M378E, ATP-synthesis, pH 7.6, temperature not specified in the publication
0.18
ADP
mutant I18W/A23F/C24F, ATP-synthesis, pH 7.6, temperature not specified in the publication
0.34
ADP
mutant L648E, ATP-synthesis, pH 7.6, temperature not specified in the publication
0.42
ADP
mutant L720E, ATP-synthesis, pH 7.6, temperature not specified in the publication
0.53
ADP
mutant A23F, ATP-synthesis, pH 7.6, temperature not specified in the publication
0.013
ATP
mutant G575K, HCO3-dependent ATPase activity, pH 7.6, temperature not specified in the publication
0.014
ATP
mutant G575F, Gln-dependent ATPase activity, pH 7.6, temperature not specified in the publication
0.014
ATP
mutant G575F, HCO3-dependent ATPase activity, pH 7.6, temperature not specified in the publication
0.03
ATP
mutant M378E, HCO3-dependent ATPase activity, pH 7.6, temperature not specified in the publication
0.038
ATP
-
pH 7.6, wild-type enzyme in presence of ornithine and in comparison to mutant enzymes
0.051
ATP
wild-type, HCO3-dependent ATPase activity, pH 7.6, temperature not specified in the publication
0.07
ATP
mutant G575K, Gln-dependent ATPase activity, pH 7.6, temperature not specified in the publication
0.12
ATP
mutant A23F, HCO3-dependent ATPase activity, pH 7.6, temperature not specified in the publication
0.16
ATP
mutant M378E, Gln-dependent ATPase activity, pH 7.6, temperature not specified in the publication
0.21
ATP
mutant I18W/A23F/C24F, HCO3-dependent ATPase activity, pH 7.6, temperature not specified in the publication
0.22
ATP
mutant L720E, HCO3-dependent ATPase activity, pH 7.6, temperature not specified in the publication
0.24
ATP
wild-type, Gln-dependent ATPase activity, pH 7.6, temperature not specified in the publication
0.25
ATP
mutant I18W/A23F/C24F, Gln-dependent ATPase activity, pH 7.6, temperature not specified in the publication
0.26
ATP
mutant M911E, HCO3-dependent ATPase activity, pH 7.6, temperature not specified in the publication
0.28
ATP
mutant M174E/M378E, Gln-dependent ATPase activity, pH 7.6, temperature not specified in the publication
0.29
ATP
mutant M174E/M378E, HCO3-dependent ATPase activity, pH 7.6, temperature not specified in the publication
0.33
ATP
mutant A23K, HCO3-dependent ATPase activity, pH 7.6, temperature not specified in the publication
0.36
ATP
mutant M174E, HCO3-dependent ATPase activity, pH 7.6, temperature not specified in the publication
0.37
ATP
-
pH 7.6, wild-type enzyme in presence of IMP and in comparison to mutant enzymes
0.38
ATP
mutant A23K, Gln-dependent ATPase activity, pH 7.6, temperature not specified in the publication
0.46
ATP
-
chimeric enzyme, in comparison to effect of phosphorylation and presence of effectors
0.47
ATP
-
pH 7.6, wild-type enzyme in absence of effectors and in comparison to mutant enzymes
0.5
ATP
mutant L648E, HCO3-dependent ATPase activity, pH 7.6, temperature not specified in the publication
0.69
ATP
mutant M174E, Gln-dependent ATPase activity, pH 7.6, temperature not specified in the publication
1
ATP
mutant M911E, Gln-dependent ATPase activity, pH 7.6, temperature not specified in the publication
1.2
ATP
mutant L648E, Gln-dependent ATPase activity, pH 7.6, temperature not specified in the publication
1.6
ATP
-
pH 7.6, wild-type enzyme in presence of UMP and in comparison to mutant enzymes
1.6
ATP
mutant A23F, Gln-dependent ATPase activity, pH 7.6, temperature not specified in the publication
1.7
ATP
-
HCO3-, at 2.0% dimethyl sulfoxide plus 0.3% glycerol or 8.0% glycerol
1.7
ATP
mutant L720E, Gln-dependent ATPase activity, pH 7.6, temperature not specified in the publication
1.2
HCO3-
-
pH 7.5, 37°C, value of wild-type enzyme in comparison to values of mutant enzymes
0.1
L-Gln
mutant A23K, glutaminase activity pH 7.6, temperature not specified in the publication
0.15
L-Gln
wild-type, glutaminase activity pH 7.6, temperature not specified in the publication
0.16
L-Gln
mutant I18W/A23F/C24F, glutaminase activity pH 7.6, temperature not specified in the publication
0.17
L-Gln
mutant G575K, glutaminase activity pH 7.6, temperature not specified in the publication
0.18
L-Gln
mutant L720E, glutaminase activity pH 7.6, temperature not specified in the publication
0.18
L-Gln
mutant M911E, glutaminase activity pH 7.6, temperature not specified in the publication
0.19
L-Gln
mutant G575F, glutaminase activity pH 7.6, temperature not specified in the publication
0.21
L-Gln
mutant A23F, glutaminase activity pH 7.6, temperature not specified in the publication
0.23
L-Gln
mutant L648E, glutaminase activity pH 7.6, temperature not specified in the publication
0.35
L-Gln
mutant M378E, glutaminase activity pH 7.6, temperature not specified in the publication
0.39
L-Gln
mutant M174E, glutaminase activity pH 7.6, temperature not specified in the publication
0.47
L-Gln
mutant M174E/M378E, glutaminase activity pH 7.6, temperature not specified in the publication
22
L-Gln
-
carbamoyl-phosphate synthase belonging to the arginine pathway
0.12
L-glutamine
-
pH 7.5, 37°C, value of wild-type enzyme in comparison to values of mutant enzymes
0.23
L-glutamine
-
wild type enzyme, in 50 mM HEPES (pH 7.5), at 37°C
0.26
L-glutamine
-
mutant enzyme T970A, in 50 mM HEPES (pH 7.5), at 37°C
0.28
L-glutamine
-
mutant enzyme T941F, in 50 mM HEPES (pH 7.5), at 37°C
0.3
L-glutamine
-
mutant enzyme K986I, in 50 mM HEPES (pH 7.5), at 37°C
0.361
L-glutamine
-
chimeric enzyme, in presence of ornithine and absence of UMP
0.45
L-glutamine
-
mutant enzyme T941F/K986I, in 50 mM HEPES (pH 7.5), at 37°C
0.5
L-glutamine
-
mutant enzyme T941F/T970A/K986I, in 50 mM HEPE (pH 7.5), at 37°C
0.89
L-glutamine
-
Escherichia coli enzyme, in presence of ornithine and absence of UMP
1.49
L-glutamine
-
Escherichia coli enzyme, in absence of ornithine and presence of UMP
1.86
L-glutamine
-
chimeric enzyme, in absence of ornithine and presence of UMP
9.8
NH4+
-
carbamoyl-phosphate synthase belonging to the pyrimidine pathway
additional information
additional information
-
effect of cryoprotectants on the kinetic parameters
-
additional information
additional information
-
effect of Orn or UMP on Km-values of mutant and wild type enzyme
-
additional information
additional information
-
kinetic constants of mutant and wild type enzymes
-
additional information
additional information
-
kinetic constants of mutant and wild type enzymes
-
additional information
additional information
-
Km values for the formation of ADP, glutamate and carbonyl phosphate of the wild-type and mutant enzymes, Km values for the hydrolysis of gamma-glutamyl hydroxamate, gamma-glutamyl hydrazide, gamma-glutamyl hydroxamate and hydroxylamine of the wild-type and mutant enzymes
-
additional information
additional information
-
comparison of KM of partial reactions of wild-type and mutant enzymes
-
additional information
additional information
-
comparison of Km-values for wild-type and mutant enzymes in absence and presence of ornithine
-
additional information
additional information
-
kinetics of wild-type and mutant enzymes in L-glutamine-dependent and bicarbonate-dependent ATPase activities, overview
-
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.01
ADP
mutant M911E, value below 0.01, ATP-synthesis, pH 7.6, temperature not specified in the publication
0.024
ADP
mutant L648E, ATP-synthesis, pH 7.6, temperature not specified in the publication
0.031
ADP
mutant L720E, ATP-synthesis, pH 7.6, temperature not specified in the publication
0.058
ADP
mutant G575F, ATP-synthesis, pH 7.6, temperature not specified in the publication
0.086
ADP
mutant I18W/A23F/C24F, ATP-synthesis, pH 7.6, temperature not specified in the publication
0.1
ADP
mutant M174E, ATP-synthesis, pH 7.6, temperature not specified in the publication
0.13
ADP
wild-type, ATP-synthesis, pH 7.6, temperature not specified in the publication
0.15
ADP
mutant G575K, ATP-synthesis, pH 7.6, temperature not specified in the publication
0.2
ADP
mutant M378E, ATP-synthesis, pH 7.6, temperature not specified in the publication
0.22
ADP
mutant M174E/M378E, ATP-synthesis, pH 7.6, temperature not specified in the publication
0.23
ADP
mutant A23F, ATP-synthesis, pH 7.6, temperature not specified in the publication
0.32
ADP
mutant A23K, ATP-synthesis, pH 7.6, temperature not specified in the publication
0.006
ATP
mutant I18W/A23F/C24F, Gln-dependent ATPase activity, pH 7.6, temperature not specified in the publication
0.01
ATP
mutant I18W/A23F/C24F, value below 0.01, CP-synthesis (co-substrate: L-Gln, HCO3-), pH 7.6, temperature not specified in the publication
0.01
ATP
mutant M174E/M378E, value below 0.01, CP-synthesis (co-substrate: L-Gln, HCO3-), pH 7.6, temperature not specified in the publication
0.01
ATP
mutant L648E,value below 0.01, CP-synthesis (co-substrate: L-Gln, HCO3-), pH 7.6, temperature not specified in the publication
0.01
ATP
mutant L720E, value below 0.01, CP-synthesis (co-substrate: L-Gln, HCO3-), pH 7.6, temperature not specified in the publication
0.012
ATP
mutant M378E, CP-synthesis (co-substrate: L-Gln, HCO3-), pH 7.6, temperature not specified in the publication
0.013
ATP
mutant M378E, HCO3-dependent ATPase activity, pH 7.6, temperature not specified in the publication
0.015
ATP
mutant M911E, value below 0.01, CP-synthesis (co-substrate: L-Gln, HCO3-), pH 7.6, temperature not specified in the publication
0.028
ATP
mutant M174E/M378E, HCO3-dependent ATPase activity, pH 7.6, temperature not specified in the publication
0.033
ATP
mutant A23F, CP-synthesis (co-substrate: L-Gln, HCO3-), pH 7.6, temperature not specified in the publication
0.035
ATP
mutant M378E, Gln-dependent ATPase activity, pH 7.6, temperature not specified in the publication
0.039
ATP
mutant I18W/A23F/C24F, HCO3-dependent ATPase activity, pH 7.6, temperature not specified in the publication
0.043
ATP
mutant M174E/M378E, Gln-dependent ATPase activity, pH 7.6, temperature not specified in the publication
0.053
ATP
mutant M174E, HCO3-dependent ATPase activity, pH 7.6, temperature not specified in the publication
0.072
ATP
mutant G575F, CP-synthesis (co-substrate: L-Gln, HCO3-), pH 7.6, temperature not specified in the publication
0.097
ATP
mutant A23F, HCO3-dependent ATPase activity, pH 7.6, temperature not specified in the publication
0.11
ATP
mutant M174E, Gln-dependent ATPase activity, pH 7.6, temperature not specified in the publication
0.16
ATP
mutant A23K, CP-synthesis (co-substrate: L-Gln, HCO3-), pH 7.6, temperature not specified in the publication
0.17
ATP
wild-type, HCO3-dependent ATPase activity, pH 7.6, temperature not specified in the publication
0.17
ATP
mutant A23K, HCO3-dependent ATPase activity, pH 7.6, temperature not specified in the publication
0.18
ATP
mutant G575K, HCO3-dependent ATPase activity, pH 7.6, temperature not specified in the publication
0.19
ATP
mutant G575F, HCO3-dependent ATPase activity, pH 7.6, temperature not specified in the publication
0.28
ATP
mutant A23K, Gln-dependent ATPase activity, pH 7.6, temperature not specified in the publication
0.33
ATP
mutant M911E, HCO3-dependent ATPase activity, pH 7.6, temperature not specified in the publication
0.39
ATP
mutant L720E, HCO3-dependent ATPase activity, pH 7.6, temperature not specified in the publication
0.5
ATP
mutant M911E, Gln-dependent ATPase activity, pH 7.6, temperature not specified in the publication
0.59
ATP
mutant L648E, HCO3-dependent ATPase activity, pH 7.6, temperature not specified in the publication
1.3
ATP
mutant A23F, Gln-dependent ATPase activity, pH 7.6, temperature not specified in the publication
1.9
ATP
wild-type, CP-synthesis (co-substrate: L-Gln, HCO3-), pH 7.6, temperature not specified in the publication
1.9
ATP
mutant G575K, CP-synthesis (co-substrate: L-Gln, HCO3-), pH 7.6, temperature not specified in the publication
1.9
ATP
mutant M174E, CP-synthesis (co-substrate: L-Gln, HCO3-), pH 7.6, temperature not specified in the publication
2
ATP
mutant L720E, Gln-dependent ATPase activity, pH 7.6, temperature not specified in the publication
3.2
ATP
mutant L648E, Gln-dependent ATPase activity, pH 7.6, temperature not specified in the publication
3.3
ATP
mutant G575F, Gln-dependent ATPase activity, pH 7.6, temperature not specified in the publication
3.3
ATP
mutant G575K, Gln-dependent ATPase activity, pH 7.6, temperature not specified in the publication
3.35
ATP
-
chimeric enzyme, in comparison to effect of phosphorylation and presence of effectors
3.6
ATP
wild-type, Gln-dependent ATPase activity, pH 7.6, temperature not specified in the publication
0.1
L-Gln
mutant M174E/M378E, glutaminase activity pH 7.6, temperature not specified in the publication
0.13
L-Gln
mutant I18W/A23F/C24F, glutaminase activity pH 7.6, temperature not specified in the publication
0.13
L-Gln
mutant M378E, glutaminase activity pH 7.6, temperature not specified in the publication
0.14
L-Gln
mutant M911E, glutaminase activity pH 7.6, temperature not specified in the publication
0.15
L-Gln
mutant M174E, glutaminase activity pH 7.6, temperature not specified in the publication
0.21
L-Gln
mutant A23K, glutaminase activity pH 7.6, temperature not specified in the publication
0.84
L-Gln
mutant L720E, glutaminase activity pH 7.6, temperature not specified in the publication
1.08
L-Gln
mutant A23F, glutaminase activity pH 7.6, temperature not specified in the publication
1.3
L-Gln
mutant L648E, glutaminase activity pH 7.6, temperature not specified in the publication
2.2
L-Gln
wild-type, glutaminase activity pH 7.6, temperature not specified in the publication
3
L-Gln
mutant G575F, glutaminase activity pH 7.6, temperature not specified in the publication
3.1
L-Gln
mutant G575K, glutaminase activity pH 7.6, temperature not specified in the publication
additional information
additional information
-
kcat values for the formation of ADP, glutamate and carbonyl phosphate of the wild-type and mutant enzymes, kcat values for the hydrolysis of gamma-glutamyl hydroxamate, gamma-glutamyl hydrazide, gamma-glutamyl hydroxamate and hydroxylamine of the wild-type and mutant enzymes
-
additional information
additional information
-
comparison of kcat of partial reactions of wild-type and mutant enzymes
-
additional information
additional information
-
comparison of kcat in absence and presence of L-ornithine, UMP and IMP for wild-type and mutant enzymes
-
additional information
additional information
-
comparison of kcat-values for wild-type and mutant enzymes in absence and presence of ornithine
-
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.005
UMP
-
pH 7.5, 37°C, value of wild-type enzyme in comparison to values of mutant enzymes
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IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0907
UMP
Bacillus amyloliquefaciens
-
mutant enzyme T941F, in 50 mM HEPES (pH 7.5), at 37°C
0.1504
UMP
Bacillus amyloliquefaciens
-
mutant enzyme T970A, in 50 mM HEPES (pH 7.5), at 37°C
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
additional information
-
the catalytic activity is dependent on the presence or absence of specific allosteric ligands but independent of the oligomeric state of the protein
additional information
-
constant specific activity of wild-type enzyme irrespective of the enzyme concentration or of the elution position of the enzyme in the gel filtration system under conditions at which different oligomeric forms predominate
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
4.2
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pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
6.7 - 8.5
-
6.7: highest activity in the pH-range 6.7-8.5, pH 7.2: 95% of the activity at pH 6.7, pH 8.5: 64% of the activity at pH 6.7
7.5 - 8.5
-
7.5: about 65% of maximal activity, 8.5: about 40% of maximal activity
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TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
37
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ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
genes carA and carB encoding the small and large subunit of the CPSase
-
-
brenda
possess 2 carbamoyl-phosphate synthases, one belonging to the arginine pathway, CPSase-A, and another belonging to the pyrimidine pathway, CPSase-P
-
-
brenda
atlantic halibut, yolk-sac larvae and adult, two enzymes: CPS II and CPS III
-
-
brenda
trifunctional carbamoyl-phosphate synthetase 2, aspartate transcarbamylase, and dihydroorotase
Q5EI67
UniProt
brenda
-
1725, 1729, 1731, 1738, 1741, 1742, 1744, 1746, 1748, 1751, 1754, 1755, 649457, 649849, 649877, 649883, 649886, 649895, 649964, 649986, 649994, 650020, 650110, 650289, 650512, 650798, 650927, 650936, 651292, 651471, 651882, 651901, 651937, 651952, 652028, 652201, 652351, 652418, 660796, 661134, 662650, 663329, 663332, 672238, 674273, 690935, 693247, 695002, 704173, 715279
-
-
brenda
and construction of a chimeric enzyme, in which the C-terminal 136 residues of Escherichia coli enzyme are replaced by the corresponding residues of Saccharomyces cerevisiae enzyme
-
-
brenda
2 carbamoyl-phosphate synthases, one specific for pyrimidine biosynthesis and the other for arginine biosynthesis
-
-
brenda
gene encodes four enzymatic activities of the pyrimidine pathway, i.e. GATase, CPSase, ATCase and DHOase
SwissProt
brenda
multifunctional protein, catalyzing the first three steps in de novo pyrimidine biosynthesis
-
-
brenda
multifunctional protein, catalyzing the first three steps in de novo pyrimidine biosynthesis
-
-
brenda
carbamoyl-phosphate-synthase/aspartate transcarbamoylase complex
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brenda
construction of a chimeric enzyme, in which the C-terminal 136 residues of Escherichia coli enzyme are replaced by the corresponding residues of Saccharomyces cerevisae enzyme
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brenda
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SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
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primary mouse hepatocytes release CPS1 during Fas ligand-induced apoptotic injury, and CPS1 is detected in hepatocytes upon hypoosmotic stress
brenda
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mammary carcinoma DMBA-1C, DMBA-5A, 205A, R3230AC, RNC-211
brenda
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about 10fold higher activity than in normal liver
brenda
additional information
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serum samples from patients with acute liver failure due to acetaminophen, Wilson disease, or ischemia show readily detectable CPS1 that i not observed in patients with chronic viral hepatitis or in control donors. CPS1 rapidly decreases to undetectable levels in sera of patients with acetaminophen-related acute liver failure who ultimately recover
brenda
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CPS1 is detected in mouse serum upon acute challenge with Fas-ligand or acetaminophen, or in sera of mice chronically fed the hepatotoxin 3,5-diethoxycarbonyl-1,4-dihydrocollidine. Half-life in serum is about 126 min
brenda
additional information
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rapidly growing tissues, e.g. fetal liver, spleen, and testis have much more carbamoyl-phosphate synthase activity than other tissues testes, e.g. lung, uterus, heart, brain and muscle. Tumors are more active than most normal tissues
brenda
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LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
coexpression of small GTPase Rheb with CAD leads to increased accumulation of CAD on lysosomes
brenda
presence in the cytosol surrounding the nucleus, plus partial localization in the lysosome
brenda
both a farnesyltransferase inhibitor that blocks membrane association of Rheb and knockdown of Rheb mislocalize CAD to the nucleus
brenda
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carbamoyl-phosphate-synthase/aspartate-transcarbamoylase complex
brenda
Highest Expressing Human Cell Lines
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GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
metabolism
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the enzyme is organized as CAD, i.e. carbamoyl-phosphate synthetase 2, aspartate transcarbamylase, and dihydroorotase, a tripartite enzyme that catalyzes the first three steps of pyrimidine biosynthesis
physiological function
physiological function
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carbamoylphosphate synthetase plays a specific physiological role in ammonium assimilation in the mesophyll and phloem for the synthesis and transport of glutamine, glutamate, and derived amino acids, overview
physiological function
carbamoylphosphate synthetase activity is essential for the optimal growth and acidification of Streptococcus thermophilus in cow milk
physiological function
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the enzyme is essential for HCV infection and replication in human liver cells
physiological function
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a single carbamoyl phosphate synthetase supplies both the pyrimidine and arginine pathways of this organism
physiological function
gene deletion mutants are complemented by arginine but not ornithine. The Cpa2 deletion mutant exhibits defects in asexual development and pathogenicity but not appressorium formation. The invasive hyphae of the mutant are restricted mainly to the primary infected cells. The mutant is unable to induce a plant defense response and has the ability to scavenge ROS during pathogen-plant interactions. The ATP grasp domain and each CPS domain are indispensable for the proper localization and full function of Cpa2
physiological function
small GTPases Rheb1 and Rheb2 bind to CAD. Binding is more pronounced with Rheb2 than with Rheb1. Rheb binds CAD in a GTP- and effector domain-dependent manner. Rheb binds at the C-terminal region of the carbamoyl-phosphate synthetase domain and not in the dihydroorotase and aspartate transcarbamoylase domains. Rheb stimulates carbamoyl-phosphate synthetase activity of CAD in vitro. Expression of Rheb leads to increased accumulation of CAD on lysosomes
physiological function
-
deletion of CPS1 results in loss of pathogenicity. The deletion mutants show slow growth rate, defects in appressorium formation and failed to develop lesions on apple leaves and fruits. Mutant strains show extreme sensitivity to high osmotic stress
physiological function
deletion of the CPA1 gene affects vegetative growth, arginine biosynthesis, and fungal pathogenicity. Dut1 interacts with Cpa1 but not Cpa2 in vivo. The Dut1-Cpa1 complex is localized at the mitochondria and regulates Cpa1-Cpa2 interaction in response to arginine
physiological function
Sulfolobus acidocaldarius DG6 / ATCC 49426
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a single carbamoyl phosphate synthetase supplies both the pyrimidine and arginine pathways of this organism
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physiological function
Pyricularia oryzae ATCC MYA-4617
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gene deletion mutants are complemented by arginine but not ornithine. The Cpa2 deletion mutant exhibits defects in asexual development and pathogenicity but not appressorium formation. The invasive hyphae of the mutant are restricted mainly to the primary infected cells. The mutant is unable to induce a plant defense response and has the ability to scavenge ROS during pathogen-plant interactions. The ATP grasp domain and each CPS domain are indispensable for the proper localization and full function of Cpa2
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physiological function
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deletion of the CPA1 gene affects vegetative growth, arginine biosynthesis, and fungal pathogenicity. Dut1 interacts with Cpa1 but not Cpa2 in vivo. The Dut1-Cpa1 complex is localized at the mitochondria and regulates Cpa1-Cpa2 interaction in response to arginine
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Show AA Sequence and Transmembrane Helices (48423 entries)
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PDB
SCOP
CATH
UNIPROT
ORGANISM
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MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
110000
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1 * 45000, beta, + 1 * 110000, alpha
117700
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1 * 41400 + 1 * 117700, the large subunit catalyses the carbamoyl phosphate synthesis from ammonia in three steps, and binds the effectors in its 15000 domain
118000
120000
133000
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1 * 40000, subunit contains the binding site for glutamine, + 1 * 133000, subunit contains the binding sites for NH4+, HCO3-, ATP, and the allosteric effectors. The light and heavy subunits are encoded by the genetically linked car A and car B genes, respectively
40000
41400
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1 * 41400 + 1 * 117700, the large subunit catalyses the carbamoyl phosphate synthesis from ammonia in three steps, and binds the effectors in its 15000 domain
42000
45000
56000
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x * 56000 (subunit N) + x * 71000 (subunit C), SDS-PAGE
63000
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CPS.A-subunit, CPS.B-subunit and glutaminase-subunit, 1* 64000 Da, 1 * 63000 Da and 1 * 45000 Da, but only the CPS.A-CPS.B-dimer has catalytic activity, SDS-PAGE
71000
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x * 56000 (subunit N) + x * 71000 (subunit C), SDS-PAGE
additional information
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the multienzyme complex of carbamoyl-phosphate synthase(glutamine), aspartate carbamoyltransferase and dihydroorotase has a MW of 870000 as determined by sedimentation equilibrium studies and 210000 by SDS-PAGE
118000
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000 + 1 * 118000, carbamoyl-phosphate synthase belonging to the pyrimidine pathway, SDS-PAGE
118000
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alpha,beta, 1 * 42000 + 1 * 118000, the ammonia tunnel is responsible for the migration of ammonia, that is produced at the active site of the small subunit and migrates to the large subunit, the carbamate tunnel connects the two active sites contained within the carboxy phosphate and carbamoyl phosphate domains of the large subunit
118000
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alpha,beta, 1 * 42000 + 1 * 118000, the effectors ornithine, IMP and UMP modulate the oligomerization of the heterodimer to higher ordered species
120000
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1 * 40000, glutaminase subunit, + 1 * 120000, synthase subunit. The synthase subunit consists of two homologous domains, CPS.A and CPS.B, which catalyze the two different ATP-dependent partial reactions involved in carbamoyl-phosphate synthesis
40000
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1 * 40000, glutaminase subunit, + 1 * 120000, synthase subunit. The synthase subunit consists of two homologous domains, CPS.A and CPS.B, which catalyze the two different ATP-dependent partial reactions involved in carbamoyl-phosphate synthesis
40000
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1 * 40000, subunit contains the binding site for glutamine, + 1 * 133000, subunit contains the binding sites for NH4+, HCO3-, ATP, and the allosteric effectors. The light and heavy subunits are encoded by the genetically linked car A and car B genes, respectively
42000
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alpha,beta, 1 * 42000 + 1 * 118000, the ammonia tunnel is responsible for the migration of ammonia, that is produced at the active site of the small subunit and migrates to the large subunit, the carbamate tunnel connects the two active sites contained within the carboxy phosphate and carbamoyl phosphate domains of the large subunit
42000
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alpha,beta, 1 * 42000 + 1 * 118000, the effectors ornithine, IMP and UMP modulate the oligomerization of the heterodimer to higher ordered species
45000
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1 * 45000, beta, + 1 * 110000, alpha
45000
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CPS.A-subunit, CPS.B-subunit and glutaminase-subunit, 1* 64000 Da, 1 * 63000 Da and 1 * 45000 Da, but only the CPS.A-CPS.B-dimer has catalytic activity, SDS-PAGE
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SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
dimer
heterodimer
heterotrimer
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CPS.A-subunit, CPS.B-subunit and glutaminase-subunit, 1* 64000 Da, 1 * 63000 Da and 1 * 45000 Da, but only the CPS.A-CPS.B-dimer has catalytic activity, SDS-PAGE
homotrimer
additional information
dimer
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1 * 40000, glutaminase subunit, + 1 * 120000, synthase subunit. The synthase subunit consists of two homologous domains, CPS.A and CPS.B, which catalyze the two different ATP-dependent partial reactions involved in carbamoyl-phosphate synthesis
dimer
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1 * 40000, subunit contains the binding site for glutamine, + 1 * 133000, subunit contains the binding sites for NH4+, HCO3-, ATP, and the allosteric effectors. The light and heavy subunits are encoded by the genetically linked car A and car B genes, respectively
dimer
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The enzyme consists of an amidotransferase domain or subunit, GLN, 40000 Da, that hydrolyzes glutamine and transfers the ammonia to the synthetase component, CPS, 120000 Da, where the biosynthetic reaction occurs. The CPS domain is composed of two homologous subdomains, CPS.A and CPS.B, that catalyse different ATP-dependent reactions involved in the carbamoyl phosphate synthesis. The CPS.A and CPS.B monomers could each catalyse all of the partial reactions, but catalysis of the overall synthesis of carbamoyl phosphate requires a dimer composed of two of the equivalent domains.
dimer
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1 * 41400 + 1 * 117700, the large subunit catalyses the carbamoyl phosphate synthesis from ammonia in three steps, and binds the effectors in its 15000 domain
dimer
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1 * 41400 + 1 * 117700, the large subunit catalyses the carbamoyl phosphate synthesis from ammonia in three steps, and binds the effectors in its 15000 domain
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dimer
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000 + 1 * 118000, carbamoyl-phosphate synthase belonging to the pyrimidine pathway, SDS-PAGE
dimer
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1 * 45000, beta, + 1 * 110000, alpha
heterodimer
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small and large subunit, the small subunit contains a class I (trpG)-type glutaminase domain to hydrolyze glutamine to ammonia. carB is composed of three exons encoding a 120 kDa large subunit, consisting of the duplicated synthetase regions and the ATP-binding domains to synthesize carbamoyl phosphate
heterodimer
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alpha,beta, that readily converts to an tetrameric species: 4 * alpha, beta in presence of positive allosteric effectors, the smaller of two subunits contains 382 amino acid residues and is responsible for the hydrolysis of free ammonia, the larger subunit contains 1073 amino acid residues and provides the binding sites for the two MgATP2 molecules required for the assembly of carbamoyl phosphate, structure and reaction mechanism
heterodimer
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alpha,beta, that readly coverts to an tetrameric species: 4 * alpha, beta in presence of positive allosteric effectors
heterodimer
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alpha,beta, 1 * 42000 + 1 * 118000, the effectors ornithine, IMP and UMP modulate the oligomerization of the heterodimer to higher ordered species
heterodimer
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alpha,beta, 1 * 42000 + 1 * 118000, the ammonia tunnel is responsible for the migration of ammonia, that is produced at the active site of the small subunit and migrates to the large subunit, the carbamate tunnel connects the two active sites contained within the carboxy phosphate and carbamoyl phosphate domains of the large subunit
heterodimer
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alpha,beta, the heteodimer aggregates to form tetramers of dimers, structure
heterodimer
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the small subunit belongs to the class I amidotransferase family of enzymes, the reaction mechanism of the small subunit proceeds through the formation of the glutamyl thioester intermediate
heterodimer
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alpha,beta, the small subunit is able to catalyze the hydrolysis of glutamine, the large subunit catalyses the formation of carbamoyl phosphate using ammonia as a nitrogen source
additional information
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enzyme can exist in different monomer conformations and states of association
additional information
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reversible concentration-dependent self-association. When the enzyme concentration is lower than 0.01 g/L the enzyme exists as monomer under all conditions
additional information
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enzyme exists in monomer, dimer, and higher oligomeric forms, which are associated with its regulation by allosteric effectors. In barbital buffer the enzyme is present as a monomer. In sodium phosphate buffer, the enzyme exists as a partially dissociating dimer. In presence of phosphate and a positive allosteric effector, the maximum association state of the enzyme is a tetramer. UMP promotes conversion to a dimer
additional information
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NH4+ or Orn promotes oligomer formation. Tetramer or higher oligomeric species in potassium phosphate buffer containing Orn
additional information
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in absence of effectors the enzyme behaves as a dissociating system possibly reflecting the dimer-tetramer equilibrium
additional information
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the enzyme consists of two polypeptide chains referred to as small and large subunits, which contain a total of three separate active sites that are connected by an intramolecular tunnel
additional information
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the smaller subunit contains the binding site for the hydrolysis of glutamine, whereas the large subunit catalyzes the formation of carbamoyl phosphate. The enzyme posssesses three distinct active sites and an ammonia tunnel, that starts at the active site in the small subunit and leads to the active site in the N-terminal domain of the large subunit
additional information
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crystal structure modelling and analysis, A2 is the glutamine amidotransferase domain and A1 is involved in communicating active site occupancy between the amidotransferase and synthetase active sites. Domains B and C are regions of internal duplication andeach contains an ATP grasp fold. Domains D' and D contain the interfaces for eCPS self-association. In addition to this role, domain D is the site for allosteric regulation of eCPS. The intramolecular tunnel connecting the three active sites
additional information
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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
additional information
Escherichia coli B / ATCC 11303
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enzyme exists in monomer, dimer, and higher oligomeric forms, which are associated with its regulation by allosteric effectors. In barbital buffer the enzyme is present as a monomer. In sodium phosphate buffer, the enzyme exists as a partially dissociating dimer. In presence of phosphate and a positive allosteric effector, the maximum association state of the enzyme is a tetramer. UMP promotes conversion to a dimer
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additional information
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the homologous CPS.A. and CPS.B. subdomains are functionally equivalent, although in the native enzyme they may have different functions resulting from their juxtaposition relative to the other components in the complex
additional information
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self-association does not play a role in regulation of enzyme activity
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POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
side-chain modification
side-chain modification
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construction of a chimeric enzyme consisting of Escherichia coli subdomains A1 and A2 fused to the mammalian enzyme subdomain B3
side-chain modification
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construction of a chimeric enzyme, in which the C-terminal 136 residues of Escherichia coli enzyme are replaced by the corresponding residues of Saccharomyces cerevisae enzyme
side-chain modification
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formation of a hybrid complex consisting of the mammalian glutaminase domain and the isolated Escherichia coli carbamoyl-phosphate synthetase domains, the steady state kinetic parameters of the hybrid are similar to those obtained for carbamoyl-phosphate synthase-aspartate carbamoyltransferase-dihydroorotase
side-chain modification
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construction of a chimeric enzyme, in which the C-terminal 136 residues of Escherichia coli enzyme are replaced by the corresponding residues of Saccharomyces cerevisae enzyme
side-chain modification
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construction of a chimeric enzyme, in which the C-terminal 136 residues of Escherichia coli enzyme are replaced by the corresponding residues of Saccharomyces cerevisae enzyme
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CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
C248D mutant of small subunit, crystals are grown at 4°C by batch from 8% poly(ethylene glycol) 8000, 0.65 M tetraethylammonium chloride, 0.5 mM MnCl2, 100 mM KCl, 1.5 mM ADP, 25 mM GEPES, pH 7.4, 0.5 mM L-Orn. The crystals belong to the space group P2(1)2(1)2(1) with unit cell dimensions of a = 151.1 A, b = 164.2 A, and c = 331.5A and one complete (alphabeta)4-heeterotetramer per asymmetric unit
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in presence of both IMP and ornithine, structural analysis of the nucleotide monophosphate allosteric binding site for enzyme
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molecular dynamics simulation. Carbamate, the product of the reaction involving ATP, bicarbonate, and ammonia, must be delivered from the site of formation to the site of utilization by traveling nearly 40 A within the enzyme. The tunnel is composed of three continuous water pockets and two narrow connecting parts, near residues A23 and G575. The two narrow parts render two free energy barriers of 6.7 and 8.4 kcal/mol, respectively. Three water pockets are filled with about 21, 9, and 9 waters, respectively, and the corresponding relative free energies of carbamate residing in these free energy minima are 5.8, 0, and 1.6 kcal/mol, respectively. The release of phosphate into solution at the site for the formation of carbamate allows the side chain of R306 to rotate toward E25, E383, and E604. This rotation is virtually prohibited by a barrier of at least 23 kcal/mol when phosphate remains bound. This conformational change not only opens the entrance of the tunnel but also shields the charge-charge repulsion from the three glutamate residues when carbamate passes through the tunnel
the structure of enzyme cocrystallized in the presence of the 5-adenylylimidodiphosphate, is determined to 2.1 A by X-ray crystallographic analysis, the three active sites of enzyme communicate via domain movements, the first example of such a domain movement is described
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the three active sites within the alpha,beta heterodimer are separated by a linear distance of nearly 100 A and are interconnected by two molecular tunnels
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X-ray crystallographic analysis, the three active sites on the protein are widely separated from one another
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PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
K986I
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the substrate kinetics (Km for L-glutamine) increases nearly 1.3fold compared to the wild type enzyme
T941F
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the substrate kinetics (Km for L-glutamine) increases nearly 1.2fold compared to the wild type enzyme
T941F/K986I
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the substrate kinetics (Km for L-glutamine) increases nearly 1.96fold compared to the wild type enzyme
T941F/T970A/K986I
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the substrate kinetics (Km for L-glutamine) increases nearly 2.17fold compared to the wild type enzyme
T970A
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the substrate kinetics (Km for L-glutamine) increases nearly 1.1fold compared to the wild type enzyme
K986I
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the substrate kinetics (Km for L-glutamine) increases nearly 1.3fold compared to the wild type enzyme
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T941F
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the substrate kinetics (Km for L-glutamine) increases nearly 1.2fold compared to the wild type enzyme
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T941F/K986I
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the substrate kinetics (Km for L-glutamine) increases nearly 1.96fold compared to the wild type enzyme
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T941F/T970A/K986I
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the substrate kinetics (Km for L-glutamine) increases nearly 2.17fold compared to the wild type enzyme
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T970A
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the substrate kinetics (Km for L-glutamine) increases nearly 1.1fold compared to the wild type enzyme
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K954E
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enzyme activity in absence of effectors is slightly lower when compared with the enzyme from wild-type. UTP inhibition is reduced
S974A
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under conditions of saturating Utp (2 mM), mutant enzyme shows similar degree of inhibition as wild-type enzyme. Activation by 5-phosphoribosyl-alpha-pyrophosphate is completely abolished at 0.05 mM, a concentration where near maximal activation is observed with wild-type enzyme
P1850T
Q5EI67
mutation affects a conserved residue in the linker region of Cad. Mutant animals display reduced lymphatic vessel development and hyperbranching arteries, reminiscent of Notch pathway mutants. Notch signaling is significantly reduced in the mutants
A126M
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mutation in large subunit significantly decreases synthesis of carbamoyl phosphate without completely inactivating the enzyme
A182V
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reduced apparent affinity for HCO3-, sensitivity toward UMP is unchanched in comparison to wild-type enzyme
A182V/S948F
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mutant is insensitive towards pyrimidine and purine nucleosides, activation by ornithine, although the affinity for this ligand is fivefold reduced in comparison to wild-type enzyme
A23F
A23K
A23K mutation decreases the glutamine-dependent ATPase activity by an order of magnitude. While there is a decrease in the rate of carbamoyl phosphate formation, the enzyme utilizes two molecules of ATP for every molecule of carbamoyl phosphate synthesized
A251C
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site-directed mutagenesis in the ammonia tunnel, analysis of secondary structure by circular dichroism measurements
A309C/S35C
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kinetic properties are similar to those of the wild-type enzyme
A311L
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site-directed mutagenesis in the ammonia tunnel, analysis of secondary structure by circular dichroism measurements
A314C
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site-directed mutagenesis in the ammonia tunnel, analysis of secondary structure by circular dichroism measurements
C232G/A251G/A314G
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site-directed mutagenesis in the ammonia tunnel, analysis of secondary structure by circular dichroism measurements
C232V/A251V/A314V
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site-directed mutagenesis in the ammonia tunnel, analysis of secondary structure by circular dichroism measurements
C248D
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partial glutaminase activity of the mutant protein is increased 40fold relative to the wild-type enzyme
C269G
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Cys269Gly and Cys269Ser mutants bind significant amounts of Gln but do not hydrolyze Gln. The mutants are able to catalyze carbamoyl-phosphate formation with NH4+ as nitrogen donor, at a rate equal to that of the wild type enzyme. The mutant enzyme catalyzes ATP synthesis from ADP and carbamoyl phosphate at the usual rates. Substantial increase in bicarbonate-dependent ATPase
C269S
D207A
D334A
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site-directed mutagenesis, the mutation has essentially no effect on the Km for L-glutamine
D362A
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mutation in alpha subunit, 2fold increase in turnover number for NH4+, 5.4fold decrease in KM-value for NH4+, 1.7fold increase in turnover number for Gln, 1.7fold increase in KM-value for Gln
D362A/betaR265A
-
mutation S362A in alpha-subunit, mutation R265A in beta-subunit,3fold increase in turnover number for NH4+, 2.2fold decrease in KM-value for NH4+, 1.2fold decrease in turnover number for Gln, 71fold increase in KM-value for Gln
D753X
-
residues within the carbamate domain of the large subunit of CPS are selected as targets for mutagenesis. Mutant enzymes E761A, E841Q, N843D, R845Q have diminished ability to synthesize carbamoyl phosphate. Mutant enzymes R715A, Q829A, and R675A show elevated Michaelis constants for MgATP2- in the partial back reaction. The mutant enzymes E761A, N827A, E841Q, N843D, and R845Q show significant increases in the Michaelis constant for either bicarbonate or carbamoyl phosphate. No significant alterations are noted upon mutation of either R571 or D753
DELTA119
-
truncation mutant DELTA14, with a removal of 14 amino acids at the carboxy terminus of the large subunit shows a 40fold decrease in Gln-dependent ATPase activity. Similar losses in activity are observed for the DELTA50, DELTA65, DELTA91, and DELTA119 mutant proteins. However formation of carbamoyl phosphate is detected even after the deletion of 119 amino acids from the carboxy-terminal end of the large subunit. No allosteric effects are observed in the presence of Orn even after the removal of the last 119 amino acids from the large subunit of CPS. Mutant enzymes G921V and G921I are unstable and are defective for the synthesis of carbamoyl phosphate. The T977A mutant is not regulated by UMP, but the full allosteric effect is observed with Orn. The R1030A and R1031A mutants enzymes exhibit wild type properties, mutant G921A shows no alteration in any of the allosteric properties. Mutant N1015A cannot be purified
DELTA14
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truncation mutant DELTA14, with a removal of 14 amino acids at the carboxy terminus of the large subunit shows a 40fold decrease in Gln-dependent ATPase activity. Similar losses in activity are observed for the DELTA50, DELTA65, DELTA91, and DELTA119 mutant proteins. However formation of carbamoyl phosphate is detected even after the deletion of 119 amino acids from the carboxy-terminal end of the large subunit. No allosteric effects are observed in the presence of Orn even after the removal of the last 119 amino acids from the large subunit of CPS. Mutant enzymes G921V and G921I are unstable and are defective for the synthesis of carbamoyl phosphate. The T977A mutant is not regulated by UMP, but the full allosteric effect is observed with Orn. The R1030A and R1031A mutants enzymes exhibit wild type properties, mutant G921A shows no alteration in any of the allosteric properties. Mutant N1015A cannot be purified
DELTA50
-
truncation mutant DELTA14, with a removal of 14 amino acids at the carboxy terminus of the large subunit shows a 40fold decrease in Gln-dependent ATPase activity. Similar losses in activity are observed for the DELTA50, DELTA65, DELTA91, and DELTA119 mutant proteins. However formation of carbamoyl phosphate is detected even after the deletion of 119 amino acids from the carboxy-terminal end of the large subunit. No allosteric effects are observed in the presence of Orn even after the removal of the last 119 amino acids from the large subunit of CPS. Mutant enzymes G921V and G921I are unstable and are defective for the synthesis of carbamoyl phosphate. The T977A mutant is not regulated by UMP, but the full allosteric effect is observed with Orn. The R1030A and R1031A mutants enzymes exhibit wild type properties, mutant G921A shows no alteration in any of the allosteric properties. Mutant N1015A cannot be purified
DELTA65
-
truncation mutant DELTA14, with a removal of 14 amino acids at the carboxy terminus of the large subunit shows a 40fold decrease in Gln-dependent ATPase activity. Similar losses in activity are observed for the DELTA50, DELTA65, DELTA91, and DELTA119 mutant proteins. However formation of carbamoyl phosphate is detected even after the deletion of 119 amino acids from the carboxy-terminal end of the large subunit. No allosteric effects are observed in the presence of Orn even after the removal of the last 119 amino acids from the large subunit of CPS. Mutant enzymes G921V and G921I are unstable and are defective for the synthesis of carbamoyl phosphate. The T977A mutant is not regulated by UMP, but the full allosteric effect is observed with Orn. The R1030A and R1031A mutants enzymes exhibit wild type properties, mutant G921A shows no alteration in any of the allosteric properties. Mutant N1015A cannot be purified
DELTA91
-
truncation mutant DELTA14, with a removal of 14 amino acids at the carboxy terminus of the large subunit shows a 40fold decrease in Gln-dependent ATPase activity. Similar losses in activity are observed for the DELTA50, DELTA65, DELTA91, and DELTA119 mutant proteins. However formation of carbamoyl phosphate is detected even after the deletion of 119 amino acids from the carboxy-terminal end of the large subunit. No allosteric effects are observed in the presence of Orn even after the removal of the last 119 amino acids from the large subunit of CPS. Mutant enzymes G921V and G921I are unstable and are defective for the synthesis of carbamoyl phosphate. The T977A mutant is not regulated by UMP, but the full allosteric effect is observed with Orn. The R1030A and R1031A mutants enzymes exhibit wild type properties, mutant G921A shows no alteration in any of the allosteric properties. Mutant N1015A cannot be purified
E215A
-
mutations of the residues E215A, N283A, E299Q, N301D, and R303Q result in proteins which are unable to synthesize carbamoyl phosphate at a significant rate. The binding of bicarbonate is most affected by the mutagenesis of residues E215A, E299Q, N301D, and R303Q. The Km for ATP is most affected in the mutant enzymes R129A, R169A, Q285A, and N301D. No significant changes in the catalytic constants are observed in the mutants R82A and D207A. All of the mutations, with the exception of the N301D mutant, primarily effect the enzyme by altering the step for the phosphorylation of bicarbonate. Mutation N301D also disrupts the catalytic step involved in the phosphorylation of carbamate
E25Q/E383Q
-
46.7fold decreased turnover number for carbamoyl-phosphate synthesis
E25Q/E383Q/E604Q
-
more than 700fold decreased turnover number for carbamoyl-phosphate synthesis
E299Q
-
mutations of the residues E215A, N283A, E299Q, N301D, and R303Q result in proteins which are unable to synthesize carbamoyl phosphate at a significant rate. The binding of bicarbonate is most affected by the mutagenesis of residues E215A, E299Q, N301D, and R303Q. The Km for ATP is most affected in the mutant enzymes R129A, R169A, Q285A, and N301D. No significant changes in the catalytic constants are observed in the mutants R82A and D207A. All of the mutations, with the exception of the N301D mutant, primarily effect the enzyme by altering the step for the phosphorylation of bicarbonate. Mutation N301D also disrupts the catalytic step involved in the phosphorylation of carbamate
E383Q/E916Q
-
3.9fold decreased turnover number for carbamoyl-phosphate synthesis
E577Q
-
437.5fold decreased turnover number for carbamoyl-phosphate synthesis
E761A
E783A
-
the allosteric activation of enzyme by ornithine is completely suppressed
E783K
-
the allosteric activation of enzyme by ornithine is completely suppressed
E841K
-
comparison of 15N-isotope effects in mutant and wild-type enzyme on the hydrolysis of glutamine, the rate of glutamine hydrolysis in the mutant is not affected by MgATP2 and HCO3-, with the wild-type enzyme in the absence of MgATP2 and HCO3- the isotope effect id reduced
E841Q
-
residues within the carbamate domain of the large subunit of CPS are selected as targets for mutagenesis. Mutant enzymes E761A, E841Q, N843D, R845Q have diminished ability to synthesize carbamoyl phosphate. Mutant enzymes R715A, Q829A, and R675A show elevated Michaelis constants for MgATP2- in the partial back reaction. The mutant enzymes E761A, N827A, E841Q, N843D, and R845Q show significant increases in the Michaelis constant for either bicarbonate or carbamoyl phosphate. No significant alterations are noted upon mutation of either R571 or D753
E892A
-
the allosteric activation of enzyme by ornithine is completely suppressed
E892K
-
the allosteric activation of enzyme by ornithine is completely suppressed
G1008A
-
mutation abolishs IMP activation and UMP inhibition in comparison to wild-type enzyme
G359F
G359Y
-
glutaminase and bicarbonate-dependent ATPase reaction are uncoupled from one another, the mutant enzyme is fully functional when external ammonia is utilized as the nitrogen source but is unable to use glutamine for the synthesis of carbamoyl phosphate
G575F
G575K
mutation to G575 does not exhibit significant pertubations to the kinetic constants of the partial reactions, mutant has similar catalytic properties as the wild-type protein, suggesting that the conformational change of Arg-848 may not be crucial for the transport of carbamate
G824D
-
strongly reduced affinity for ornithine in comparison to wild-type enzyme
G919C
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
G921A
-
truncation mutant DELTA14, with a removal of 14 amino acids at the carboxy terminus of the large subunit shows a 40fold decrease in Gln-dependent ATPase activity. Similar losses in activity are observed for the DELTA50, DELTA65, DELTA91, and DELTA119 mutant proteins. However formation of carbamoyl phosphate is detected even after the deletion of 119 amino acids from the carboxy-terminal end of the large subunit. No allosteric effects are observed in the presence of Orn even after the removal of the last 119 amino acids from the large subunit of CPS. Mutant enzymes G921V and G921I are unstable and are defective for the synthesis of carbamoyl phosphate. The T977A mutant is not regulated by UMP, but the full allosteric effect is observed with Orn. The R1030A and R1031A mutants enzymes exhibit wild type properties, mutant G921A shows no alteration in any of the allosteric properties. Mutant N1015A cannot be purified
G921I
-
truncation mutant DELTA14, with a removal of 14 amino acids at the carboxy terminus of the large subunit shows a 40fold decrease in Gln-dependent ATPase activity. Similar losses in activity are observed for the DELTA50, DELTA65, DELTA91, and DELTA119 mutant proteins. However formation of carbamoyl phosphate is detected even after the deletion of 119 amino acids from the carboxy-terminal end of the large subunit. No allosteric effects are observed in the presence of Orn even after the removal of the last 119 amino acids from the large subunit of CPS. Mutant enzymes G921V and G921I are unstable and are defective for the synthesis of carbamoyl phosphate. The T977A mutant is not regulated by UMP, but the full allosteric effect is observed with Orn. The R1030A and R1031A mutants enzymes exhibit wild type properties, mutant G921A shows no alteration in any of the allosteric properties. Mutant N1015A cannot be purified
G921V
-
truncation mutant DELTA14, with a removal of 14 amino acids at the carboxy terminus of the large subunit shows a 40fold decrease in Gln-dependent ATPase activity. Similar losses in activity are observed for the DELTA50, DELTA65, DELTA91, and DELTA119 mutant proteins. However formation of carbamoyl phosphate is detected even after the deletion of 119 amino acids from the carboxy-terminal end of the large subunit. No allosteric effects are observed in the presence of Orn even after the removal of the last 119 amino acids from the large subunit of CPS. Mutant enzymes G921V and G921I are unstable and are defective for the synthesis of carbamoyl phosphate. The T977A mutant is not regulated by UMP, but the full allosteric effect is observed with Orn. The R1030A and R1031A mutants enzymes exhibit wild type properties, mutant G921A shows no alteration in any of the allosteric properties. Mutant N1015A cannot be purified
G997A
-
mutation abolishs IMP activation and UMP inhibition in comparison to wild-type enzyme
H975L
-
mutation abolishes oligomer formation even high enzyme concentrations or in the presence of ornithine
H975L/N987V
-
mutation abolishes oligomer formation even high enzyme concentrations or in the presence of ornithine
H995A
I18W/A23F/C24F
triple mutant I18W/A23F/C24F is made to disrupt the water pocket that may facilitate the passage of carbamate through the carbamate tunnel. This mutant significantly hinders the overall rate of carbamoyl phosphate synthesis and it diminishes all of the other partial reactions
I352F
-
site-directed mutagenesis in the ammonia tunnel, analysis of secondary structure by circular dichroism measurements
K1061A
-
mutation abolishs IMP activation and UMP inhibition in comparison to wild-type enzyme
K954A
-
mutation has little effect on enzyme activity in comparison to wild-type enzyme
K993W
-
mutation has little effect on enzyme activity in comparison to wild-type enzyme
K993W/H995A
-
mutation abolishs IMP activation and UMP inhibition in comparison to wild-type enzyme
L270K
-
ammonia-dependent carbamoyl phosphate synthesis activity is very similar to that of the wild-type enzyme, L-glutamine-dependent carbamoyl phosphate synthesis activity is 5fold decreased in comparison to the wild-type enzyme, the glutamine binding is almost entirely abolished
L421E
L421E/H975L/N987V
-
mutation abolishes oligomer formation even high enzyme concentrations or in the presence of ornithine
L648E
mutant shows no detectable rate of carbamoyl phosphate formation. Little effect on the rates of all partial reactions is observed. Thus the reactions at the small subunit and the carboxy phosphate active sites remains unperturbed. The L648E mutant exhibits a 10fold drop in the rate of the glutaminase reaction which is due to the uncoupling between the carboxy phosphate and glutaminase active sites
L720E
the rates for both the glutamine- and HCO3--dependent ATPase reactions are largely unaffected by the mutation. The rate of the partial ATP-synthesis reaction is decreased 4fold. These perturbations may be due to an altered active site environment which diminishes the rate ADP phosphorylation by carbamoyl phosphate. No carbamoyl phosphate formation is detected. Mutant can structurally block the exit of the carbamate tunnel although the presence of the glutamate also weakens the assistance of Arg-848 during the synthesis of carbamoyl phosphate
L990A
M174E
mutant has significant reductions in the rates of the ATPase and carbamoyl phosphate synthesis reactions. Mutation does not affect the rate of the partial ATP synthesis reaction
M174E/M378E
catalytic properties of the double mutant, M174E/M378E, are similar to the single mutants with regard to the various partial reactions. Double mutant is unable to synthesize carbamoyl phosphate
M378E
mutant has significant reductions in the rates of the ATPase and carbamoyl phosphate synthesis reactions. Mutation does not affect the rate of the partial ATP synthesis reaction
M911E
the rate for both the glutamine- and HCO3--dependent ATPase reactions are largely unaffected by the mutation. The rate of the partial ATP-synthesis reaction is virtually undetectable. This perturbation may be due to an altered active site environment which diminishes the rate ADP phosphorylation by carbamoyl phosphate. Only limited carbamoyl phosphate formation is detected with mutant M911E. M911E mutant can structurally block the exit of the carbamate tunnel although the presence of glutamate also weakens the assistance of Arg-848 during the synthesis of carbamoyl phosphate
N1015A
-
truncation mutant DELTA14, with a removal of 14 amino acids at the carboxy terminus of the large subunit shows a 40fold decrease in Gln-dependent ATPase activity. Similar losses in activity are observed for the DELTA50, DELTA65, DELTA91, and DELTA119 mutant proteins. However formation of carbamoyl phosphate is detected even after the deletion of 119 amino acids from the carboxy-terminal end of the large subunit. No allosteric effects are observed in the presence of Orn even after the removal of the last 119 amino acids from the large subunit of CPS. Mutant enzymes G921V and G921I are unstable and are defective for the synthesis of carbamoyl phosphate. The T977A mutant is not regulated by UMP, but the full allosteric effect is observed with Orn. The R1030A and R1031A mutants enzymes exhibit wild type properties, mutant G921A shows no alteration in any of the allosteric properties. Mutant N1015A cannot be purified
N283A
-
mutations of the residues E215A, N283A, E299Q, N301D, and R303Q result in proteins which are unable to synthesize carbamoyl phosphate at a significant rate. The binding of bicarbonate is most affected by the mutagenesis of residues E215A, E299Q, N301D, and R303Q. The Km for ATP is most affected in the mutant enzymes R129A, R169A, Q285A, and N301D. No significant changes in the catalytic constants are observed in the mutants R82A and D207A. All of the mutations, with the exception of the N301D mutant, primarily effect the enzyme by altering the step for the phosphorylation of bicarbonate. Mutation N301D also disrupts the catalytic step involved in the phosphorylation of carbamate
N301D
-
mutations of the residues E215A, N283A, E299Q, N301D, and R303Q result in proteins which are unable to synthesize carbamoyl phosphate at a significant rate. The binding of bicarbonate is most affected by the mutagenesis of residues E215A, E299Q, N301D, and R303Q. The Km for ATP is most affected in the mutant enzymes R129A, R169A, Q285A, and N301D. No significant changes in the catalytic constants are observed in the mutants R82A and D207A. All of the mutations, with the exception of the N301D mutant, primarily effect the enzyme by altering the step for the phosphorylation of bicarbonate. Mutation N301D also disrupts the catalytic step involved in the phosphorylation of carbamate
N301K
-
mutation in large subunit significantly decreases synthesis of carbamoyl phosphate without completely inactivating the enzyme
N311A
-
site-directed mutagenesis, the mutant shows increased Km for L-glutamine compared to the wild-type enzyme
N827A
-
residues within the carbamate domain of the large subunit of CPS are selected as targets for mutagenesis. Mutant enzymes E761A, E841Q, N843D, R845Q have diminished ability to synthesize carbamoyl phosphate. Mutant enzymes R715A, Q829A, and R675A show elevated Michaelis constants for MgATP2- in the partial back reaction. The mutant enzymes E761A, N827A, E841Q, N843D, and R845Q show significant increases in the Michaelis constant for either bicarbonate or carbamoyl phosphate. No significant alterations are noted upon mutation of either R571 or D753
N843Q
-
residues within the carbamate domain of the large subunit of CPS are selected as targets for mutagenesis. Mutant enzymes E761A, E841Q, N843D, R845Q have diminished ability to synthesize carbamoyl phosphate. Mutant enzymes R715A, Q829A, and R675A show elevated Michaelis constants for MgATP2- in the partial back reaction. The mutant enzymes E761A, N827A, E841Q, N843D, and R845Q show significant increases in the Michaelis constant for either bicarbonate or carbamoyl phosphate. No significant alterations are noted upon mutation of either R571 or D753
N987D
-
mutant forms a (alpha/beta)-monomer regardless of the presence of any allosteric effectors
N992A
-
mutation abolishes oligomer formation even high enzyme concentrations or in the presence of ornithine
P165S
-
reduced apparent affinity for HCO3-, sensitivity toward UMP is increased in comparison to wild-type enzyme
P170L
-
reduced apparent affinity for HCO3-, sensitivity toward UMP is increased in comparison to wild-type enzyme
P360A/H361A
-
mutation in beta-subunit, turnover number for NH4+ is nearly identical to wild-type value, 1.6fold increase in turnover number for Gln, 3.2fold increase in KM-value for Gln
P360A/H361A/R265A
-
mutations P360A and H361A in alpha-subunit, mutation R265A in beta-subunit, mutant enzyme is unable to utilize glutamine for the synthesis of carbamoyl phosphate1.3fold increase in turnover number for NH4+, 11.8fold decrease in KM-value for NH4+
P360L
P909C
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
P909C/G919C
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Q262A/R265A
-
mutation in beta-subunit 1.9fold increase in turnover number for NH4+, 5fold decrease in KM-value for NH4+, 1.4fold decrease in turnover number for Gln, 43fold increase in KM-value for Gln
Q262A/R265A/N266A
-
mutation in beta-subunit, 1.6fold decrease in turnover number for Gln, 13.5fold increase in KM-value for Gln
Q262P
-
mutation causes marked enzyme instability, mutation in large subunit significantly decreases synthesis of carbamoyl phosphate without completely inactivating the enzyme
Q273E
-
ammonia-dependent carbamoyl phosphate synthesis activity is very similar to that of the wild-type enzyme, L-glutamine-dependent carbamoyl phosphate synthesis activity is equivalent to the wild-type enzyme, but the mutant is 10fold impaired in its L-glutamine binding ability in comparison to wild-type enzyme
Q273E/L270K
-
ammonia-dependent carbamoyl phosphate synthesis activity is very similar to that of the wild-type enzyme, L-glutamine-dependent carbamoyl phosphate synthesis activity is 25fold decreased in comparison to the wild-type enzyme, the glutamine binding is almost entirely abolished
Q273E/N240S
-
ammonia-dependent carbamoyl phosphate synthesis activity is very similar to that of the wild-type enzyme, L-glutamine-dependent carbamoyl phosphate synthesis activity is equivalent to the wild-type enzyme
Q285A
-
mutations of the residues E215A, N283A, E299Q, N301D, and R303Q result in proteins which are unable to synthesize carbamoyl phosphate at a significant rate. The binding of bicarbonate is most affected by the mutagenesis of residues E215A, E299Q, N301D, and R303Q. The Km for ATP is most affected in the mutant enzymes R129A, R169A, Q285A, and N301D. No significant changes in the catalytic constants are observed in the mutants R82A and D207A. All of the mutations, with the exception of the N301D mutant, primarily effect the enzyme by altering the step for the phosphorylation of bicarbonate. Mutation N301D also disrupts the catalytic step involved in the phosphorylation of carbamate
Q310A
-
site-directed mutagenesis, the mutant shows increased Km for L-glutamine compared to the wild-type enzyme
Q351A
-
site-directed mutagenesis, the mutant shows highly increased Km for L-glutamine compared to the wild-type enzyme
Q829A
-
residues within the carbamate domain of the large subunit of CPS are selected as targets for mutagenesis. Mutant enzymes E761A, E841Q, N843D, R845Q have diminished ability to synthesize carbamoyl phosphate. Mutant enzymes R715A, Q829A, and R675A show elevated Michaelis constants for MgATP2- in the partial back reaction. The mutant enzymes E761A, N827A, E841Q, N843D, and R845Q show significant increases in the Michaelis constant for either bicarbonate or carbamoyl phosphate. No significant alterations are noted upon mutation of either R571 or D753
R1020A
-
mutation has little effect on enzyme activity in comparison to wild-type enzyme
R1021A
-
mutation has little effect on enzyme activity in comparison to wild-type enzyme
R1030A
-
truncation mutant DELTA14, with a removal of 14 amino acids at the carboxy terminus of the large subunit shows a 40fold decrease in Gln-dependent ATPase activity. Similar losses in activity are observed for the DELTA50, DELTA65, DELTA91, and DELTA119 mutant proteins. However formation of carbamoyl phosphate is detected even after the deletion of 119 amino acids from the carboxy-terminal end of the large subunit. No allosteric effects are observed in the presence of Orn even after the removal of the last 119 amino acids from the large subunit of CPS. Mutant enzymes G921V and G921I are unstable and are defective for the synthesis of carbamoyl phosphate. The T977A mutant is not regulated by UMP, but the full allosteric effect is observed with Orn. The R1030A and R1031A mutants enzymes exhibit wild type properties, mutant G921A shows no alteration in any of the allosteric properties. Mutant N1015A cannot be purified
R1031A
-
truncation mutant DELTA14, with a removal of 14 amino acids at the carboxy terminus of the large subunit shows a 40fold decrease in Gln-dependent ATPase activity. Similar losses in activity are observed for the DELTA50, DELTA65, DELTA91, and DELTA119 mutant proteins. However formation of carbamoyl phosphate is detected even after the deletion of 119 amino acids from the carboxy-terminal end of the large subunit. No allosteric effects are observed in the presence of Orn even after the removal of the last 119 amino acids from the large subunit of CPS. Mutant enzymes G921V and G921I are unstable and are defective for the synthesis of carbamoyl phosphate. The T977A mutant is not regulated by UMP, but the full allosteric effect is observed with Orn. The R1030A and R1031A mutants enzymes exhibit wild type properties, mutant G921A shows no alteration in any of the allosteric properties. Mutant N1015A cannot be purified
R129A
-
mutations of the residues E215A, N283A, E299Q, N301D, and R303Q result in proteins which are unable to synthesize carbamoyl phosphate at a significant rate. The binding of bicarbonate is most affected by the mutagenesis of residues E215A, E299Q, N301D, and R303Q. The Km for ATP is most affected in the mutant enzymes R129A, R169A, Q285A, and N301D. No significant changes in the catalytic constants are observed in the mutants R82A and D207A. All of the mutations, with the exception of the N301D mutant, primarily effect the enzyme by altering the step for the phosphorylation of bicarbonate. Mutation N301D also disrupts the catalytic step involved in the phosphorylation of carbamate
R169A
-
mutations of the residues E215A, N283A, E299Q, N301D, and R303Q result in proteins which are unable to synthesize carbamoyl phosphate at a significant rate. The binding of bicarbonate is most affected by the mutagenesis of residues E215A, E299Q, N301D, and R303Q. The Km for ATP is most affected in the mutant enzymes R129A, R169A, Q285A, and N301D. No significant changes in the catalytic constants are observed in the mutants R82A and D207A. All of the mutations, with the exception of the N301D mutant, primarily effect the enzyme by altering the step for the phosphorylation of bicarbonate. Mutation N301D also disrupts the catalytic step involved in the phosphorylation of carbamate
R169H
-
mutation in large subunit significantly decreases synthesis of carbamoyl phosphate without completely inactivating the enzyme
R265A
-
mutation in beta-subunit, 1.3fold increase in turnover number for NH4+, 5fold decrease in KM-value for NH4+, 1.3fold decrease in turnover number for Gln, 69fold increase in KM-value for Gln
R303Q
-
mutations of the residues E215A, N283A, E299Q, N301D, and R303Q result in proteins which are unable to synthesize carbamoyl phosphate at a significant rate. The binding of bicarbonate is most affected by the mutagenesis of residues E215A, E299Q, N301D, and R303Q. The Km for ATP is most affected in the mutant enzymes R129A, R169A, Q285A, and N301D. No significant changes in the catalytic constants are observed in the mutants R82A and D207A. All of the mutations, with the exception of the N301D mutant, primarily effect the enzyme by altering the step for the phosphorylation of bicarbonate. Mutation N301D also disrupts the catalytic step involved in the phosphorylation of carbamate
R306A
-
more than 700fold decreased turnover number for carbamoyl-phosphate synthesis
R571X
-
residues within the carbamate domain of the large subunit of CPS are selected as targets for mutagenesis. Mutant enzymes E761A, E841Q, N843D, R845Q have diminished ability to synthesize carbamoyl phosphate. Mutant enzymes R715A, Q829A, and R675A show elevated Michaelis constants for MgATP2- in the partial back reaction. The mutant enzymes E761A, N827A, E841Q, N843D, and R845Q show significant increases in the Michaelis constant for either bicarbonate or carbamoyl phosphate. No significant alterations are noted upon mutation of either R571 or D753
R675A
-
residues within the carbamate domain of the large subunit of CPS are selected as targets for mutagenesis. Mutant enzymes E761A, E841Q, N843D, R845Q have diminished ability to synthesize carbamoyl phosphate. Mutant enzymes R715A, Q829A, and R675A show elevated Michaelis constants for MgATP2- in the partial back reaction. The mutant enzymes E761A, N827A, E841Q, N843D, and R845Q show significant increases in the Michaelis constant for either bicarbonate or carbamoyl phosphate. No significant alterations are noted upon mutation of either R571 or D753
R675L
-
mutation in large subunit significantly decreases synthesis of carbamoyl phosphate without completely inactivating the enzyme
R715A
-
residues within the carbamate domain of the large subunit of CPS are selected as targets for mutagenesis. Mutant enzymes E761A, E841Q, N843D, R845Q have diminished ability to synthesize carbamoyl phosphate. Mutant enzymes R715A, Q829A, and R675A show elevated Michaelis constants for MgATP2- in the partial back reaction. The mutant enzymes E761A, N827A, E841Q, N843D, and R845Q show significant increases in the Michaelis constant for either bicarbonate or carbamoyl phosphate. No significant alterations are noted upon mutation of either R571 or D753
R82A
-
mutations of the residues E215A, N283A, E299Q, N301D, and R303Q result in proteins which are unable to synthesize carbamoyl phosphate at a significant rate. The binding of bicarbonate is most affected by the mutagenesis of residues E215A, E299Q, N301D, and R303Q. The Km for ATP is most affected in the mutant enzymes R129A, R169A, Q285A, and N301D. No significant changes in the catalytic constants are observed in the mutants R82A and D207A. All of the mutations, with the exception of the N301D mutant, primarily effect the enzyme by altering the step for the phosphorylation of bicarbonate. Mutation N301D also disrupts the catalytic step involved in the phosphorylation of carbamate
R845Q
-
residues within the carbamate domain of the large subunit of CPS are selected as targets for mutagenesis. Mutant enzymes E761A, E841Q, N843D, R845Q have diminished ability to synthesize carbamoyl phosphate. Mutant enzymes R715A, Q829A, and R675A show elevated Michaelis constants for MgATP2- in the partial back reaction. The mutant enzymes E761A, N827A, E841Q, N843D, and R845Q show significant increases in the Michaelis constant for either bicarbonate or carbamoyl phosphate. No significant alterations are noted upon mutation of either R571 or D753
S35F
-
kinetic properties are similar to the wild-type enzyme, only KM-value of L-glutamine is 5fold increased
S35Y
-
site-directed mutagenesis in the ammonia tunnel, analysis of secondary structure by circular dichroism measurements
S743N
-
minor modification of kinetic parameters in comparison to wild-type enzyme
S743N/G824D
-
strongly reduced affinity for ornithine in comparison to wild-type enzyme
S789P
-
mutation in large subunit significantly decreases synthesis of carbamoyl phosphate without completely inactivating the enzyme
S948A
-
mutation has little effect on enzyme activity in comparison to wild-type enzyme
S948F
-
mutant enzyme is unsensitive to UMP and IMP, but is still activated by ornithine, although to a reduced extent
T1042I
T1043K
-
the allosteric activation of enzyme by ornithine is completely suppressed
T249V
-
site-directed mutagenesis in the ammonia tunnel, analysis of secondary structure by circular dichroism measurements
T800F
-
reduced affinity for ornithine, increased sensitivity for UMP in comparison to wild-type enzyme
T974A
-
mutation abolishs IMP activation and UMP inhibition in comparison to wild-type enzyme
T977A
-
truncation mutant DELTA14, with a removal of 14 amino acids at the carboxy terminus of the large subunit shows a 40fold decrease in Gln-dependent ATPase activity. Similar losses in activity are observed for the DELTA50, DELTA65, DELTA91, and DELTA119 mutant proteins. However formation of carbamoyl phosphate is detected even after the deletion of 119 amino acids from the carboxy-terminal end of the large subunit. No allosteric effects are observed in the presence of Orn even after the removal of the last 119 amino acids from the large subunit of CPS. Mutant enzymes G921V and G921I are unstable and are defective for the synthesis of carbamoyl phosphate. The T977A mutant is not regulated by UMP, but the full allosteric effect is observed with Orn. The R1030A and R1031A mutants enzymes exhibit wild type properties, mutant G921A shows no alteration in any of the allosteric properties. Mutant N1015A cannot be purified
V640R
-
mutation in large subunit significantly decreases synthesis of carbamoyl phosphate without completely inactivating the enzyme
V991A
-
mutation has little effect on enzyme activity in comparison to wild-type enzyme
K954A
-
mutation has little effect on enzyme activity in comparison to wild-type enzyme
-
S948A
-
mutation has little effect on enzyme activity in comparison to wild-type enzyme
-
V991A
-
mutation has little effect on enzyme activity in comparison to wild-type enzyme
-
S44A
-
mutant of hybrid enzyme shows km value similar to the wild-type hybrid, the kcat values of glutamine and ATP are 14fold lower in comparison to wild-type hybrid, the functional linkage that coordinates the reactions on the glutaminase and carbamoyl-phosphate synthetase domains is lost as a result of mutation
D1220Y
aspartate transcarbamylase activity of the multifunctional mutant protein carbamylphosphate synthetase/aspartate transcarbamylase is no longer inhibited by UTP, carbamylphosphate synthetase activity retains fully sensitive to this effector
N1094D
aspartate transcarbamylase activity of the multifunctional mutant protein carbamylphosphate synthetase/aspartate transcarbamylase is no longer inhibited by UTP, carbamylphosphate synthetase activity retains fully sensitive to this effector
R1076S
aspartate transcarbamylase activity of the multifunctional mutant protein carbamylphosphate synthetase/aspartate transcarbamylase is no longer inhibited by UTP, carbamylphosphate synthetase activity retains fully sensitive to this effector
Y1096C
aspartate transcarbamylase activity of the multifunctional mutant protein carbamylphosphate synthetase/aspartate transcarbamylase is no longer inhibited by UTP, carbamylphosphate synthetase activity retains fully sensitive to this effector
Y1112L
aspartate transcarbamylase activity of the multifunctional mutant protein carbamylphosphate synthetase/aspartate transcarbamylase is no longer inhibited by UTP, carbamylphosphate synthetase activity retains fully sensitive to this effector
C345A
-
site-directed mutagenesis of an essential catalytic triad residue of the GATase domain, the mutation completely abolishes complementation activity and GATase activity
N348A
-
site-directed mutagenesis, the mutation reduces complementation activity, GATase activity and the initial growth rate, but the N348A mutant exhibits the high virulence phenotype of the parental RH strain in C57Bl/6 mice
N348R
-
site-directed mutagenesis, the mutation completely abolishes complementation activity and GATase activity
additional information
A23F
mutant designed to block the migration of carbamate through the narrowest parts of the carbamate tunnel. Mutant retains 1.7% of the catalytic activity for the synthesis of carbamoyl phosphate relative to the wild type CPS
A23F
migration of carbamate through the narrowest part of the carbamate tunnel is blocked. From the kinetic data the only reaction significantly affected by this mutation is the overall synthesis of carbamoyl phosphate (only 1.7% compared to wild-type)
C269S
-
Cys269Gly and Cys269Ser mutants bind significant amounts of Gln but do not hydrolyze Gln. The mutants are able to catalyze carbamoyl-phosphate formation with NH4+ as nitrogen donor, at a rate equal to that of the wild type enzyme. The mutant enzyme catalyzes ATP synthesis from ADP and carbamoyl phosphate at the usual rates. Substantial increase in bicarbonate-dependent ATPase
C269S
-
mutant is unable to hydrolyse glutamine, the mutant is proved to be an adequate mimic for studies of the Michaelis complex
D207A
-
mutations of the residues E215A, N283A, E299Q, N301D, and R303Q result in proteins which are unable to synthesize carbamoyl phosphate at a significant rate. The binding of bicarbonate is most affected by the mutagenesis of residues E215A, E299Q, N301D, and R303Q. The Km for ATP is most affected in the mutant enzymes R129A, R169A, Q285A, and N301D. No significant changes in the catalytic constants are observed in the mutants R82A and D207A. All of the mutations, with the exception of the N301D mutant, primarily effect the enzyme by altering the step for the phosphorylation of bicarbonate. Mutation N301D also disrupts the catalytic step involved in the phosphorylation of carbamate
E761A
-
residues within the carbamate domain of the large subunit of CPS are selected as targets for mutagenesis. Mutant enzymes E761A, E841Q, N843D, R845Q have diminished ability to synthesize carbamoyl phosphate. Mutant enzymes R715A, Q829A, and R675A show elevated Michaelis constants for MgATP2- in the partial back reaction. The mutant enzymes E761A, N827A, E841Q, N843D, and R845Q show significant increases in the Michaelis constant for either bicarbonate or carbamoyl phosphate. No significant alterations are noted upon mutation of either R571 or D753
E761A
-
the residue is a key in the allosteric signal transduction pathway from ornithine to ATP
G359F
-
glutaminase and bicarbonate-dependent ATPase reaction are uncoupled from one another, the mutant enzyme is fully functional when external ammonia is utilized as the nitrogen source but is unable to use glutamine for the synthesis of carbamoyl phosphate
G575F
mutant designed to block the migration of carbamate through the narrowest parts of the carbamate tunnel. Mutant retains 3.8% of the catalytic activity for the synthesis of carbamoyl phosphate relative to the wild type CPS
G575F
mutation to G575 does not exhibit significant pertubations to the kinetic constants of the partial reactions, G575F mutant has a 50fold reduction in the rate of carbamoyl phosphate formation (attributed to the restricted passage of carbamate through the tunnel). The insertion of a larger phenylalanine side chain is anticipated to create a more efficient blockage of the tunnel. Of the mutants made in the carbamate tunnel, G575F is the most efficacious at blocking the passage of carbamate without disrupting any of the active sites
H995A
-
mutation has little effect on enzyme activity in comparison to wild-type enzyme
H995A
-
mutant exhibits normal kinetics for the activator ornithine and for the substrates in presence of ornithine, the mutation selectiviely decrease 5.6 and 2.3fold the apparent affinity for UMP and IMP, mutation abolishes the ability to be photoaffinity labeled with IMP
L421E
-
mutant forms a (alpha/beta)2-dimer in presence of ornithine, IMP and UMP
L990A
-
mutation abolishs IMP activation and UMP inhibition in comparison to wild-type enzyme
L990A
-
mutation abolishes oligomer formation even high enzyme concentrations or in the presence of ornithine
P360L
-
UMP still inhibits the activity of the mutant enzyme, 30fold reduced affinity for ornithine and 20fold reduced affinity for IMP in comparison to wild-type enzyme
P360L
-
mutation in large subunit significantly decreases synthesis of carbamoyl phosphate without completely inactivating the enzyme
T1042I
-
mutation reduces activation by ornithine, the mutated enzyme is still sensitive to UMP and IMP
T1042I
-
greatly reduced activation by ornithine, the affinities for both UMP and IMP are reduced in comparison to wild-type enzyme in comparison to wild-type enzyme
additional information
-
construction of carB mutants lacking a functional large subunit. The carB mutant shows negligible effects on overall arginine levels. The levels of ornithine, citrulline and arginine remain low in the mutant. the ammonium level of carB mutant cells are similar to the wild-type in air, overview
additional information
Q5EI67
mutant allele sl23 is a 20 bp deletion in exon 5 of the carbamoyl-phosphate synthetase 2-aspartate transcarbamylase-dihydroorotase corresponding to aa 170-175 of the wild-type protein. In the embryos, axons arising from the epibranchial sensory ganglia do not form their stereotypical terminal fields in the hindbrain. The embryos also have small eyes and deformed jaws. Mutants do not exhibit elevated cell death, or gross morphological changes, in their hindbrains. Interference with glycosylation via tunicamycin treatment mimicks the phenotype
additional information
supressor of black mutation, Su(b), which causes a loss of UTP feedback inhibition, is a missense mutation resulting in a glutamate to lysine substitution within the second ATP binding site of enzyme
additional information
-
smaller recombinant proteins in which the subdomains A1, A2, A3, B1, B2, or B3 of the synthetase subunit are deleted, show that subdomains A1 and B1 are attenuation domains which suppress the intrinsically high activity and are required for the physical association with the glutaminase subunit
additional information
-
the 120000 MW synthetase subunit and the 40000 MW glutaminase subunit, in which the intergenic region between the contiguous carA and carB genes is deleted and the sequence encoding the carbamoyl-phosphate synthase subunits is fused in frame. The resulting fusion protein is activated 10fold relative to the native protein, it is responsive to the allosteric activator Orn, and cannot use NH4+ as a nitrogen donor. The functional linkage that coordinates the rate of glutamine hydrolysis with the activation of bicarbonate is abolished
additional information
-
A series of mutants created within the ammonia tunnel are prepared by placement of a constriction via site-directed mutagenesis. The degree of constriction within the ammonia tunnel of these enzymes is found to correlate to the extent of the uncoupling of the partial reactions, the diminution of carbamoyl phosphate formation, and the percentage of the internally derived ammonia that is channelled through the ammonia tunnel
additional information
-
A series of mutants created and probes the direct transport of carbamate through the carbamate tunnel, the partial reactions of enzyme have not been significantly impaired by these mutations, and thus, the catalytic machinery at the individual active site has not been functionally pertured
additional information
-
a chimeric deletion mutant A1A2B3 lacks 59 of 163 residues from the carboxyl end of the Escherichia coli /mammalian chimera, this mutant is somewhat more active than the full-length chimeric protein but has reduced sensitivity to allosteric effectors
additional information
-
several mutations increase, reduce, suppresse, or reverse the allosteric effects of ornithine, UMP and IMP, the mutational analysis of ornithine, and potassium binding site strongly supports the proposal for a coupling between the allosteric transduction signal pathways for UMP,IMP and ornithine
additional information
-
the mutants are constructed to mimic the potentially significant substitutions which is observed in the frog enzyme and to allow determination of the effects of these changes on the glutamine binding ability of enzyme
additional information
-
downregulation of CAD, i.e. carbamoyl-phosphate synthetase 2, aspartate transcarbamylase, and dihydroorotase, by specific siRNA
additional information
-
the 29-amino acid linker, which connects the glutaminase domain and the synthetase domain is deleted. The deletion mutant catalyzes glutamine-dependent carbamoyl phosphate synthesis, but is activated 10fold relative to its wild type counterpart. NH4+-dependent synthesis of carbamoyl phosphate is abolished. The mutant enzyme is still sensitive to inhibition by the allosteric effector UTP, but is no longer activated by the allosteric effector phosphoribosyl diphosphate. The linker appears to serve as a spacer that allows the complex to cycle between two conformations, an open low activity form in which the NH4+ site on the synthetase domain is accessible and an activated conformation in which the NH4+ generated in situ from Gln is directly channeled to the synthetase domain active site and access to exogenous NH4+ is blocked
additional information
phenotype of deletion mutants lacking the ATP domain or GPS1 or GPS2 domain is similar to mutants lacking the entire gene. Presence of 2.5 mM exogenous arginine can restore the defects of each transformant
additional information
Pyricularia oryzae ATCC MYA-4617
-
phenotype of deletion mutants lacking the ATP domain or GPS1 or GPS2 domain is similar to mutants lacking the entire gene. Presence of 2.5 mM exogenous arginine can restore the defects of each transformant
-
additional information
construction by gene replacement and phenotype of a carbamoylphosphate synthetase-negative mutant A17, i.e. DELTAcarB, overview. CO2 depletion or carB gene inactivation determines the auxotrophy of Streptococcus thermophilus for L-arginine and uracil. The parent strain grows faster than the mutant, even when milk is supplemented with uracil or arginine
additional information
-
disruption of Toxoplasma gondii CPSII induces a severe uracil auxotrophy with no detectable parasite replication in vitro and complete attenuation of virulence in mice. A CPSII cDNA minigene efficiently complements the uracil auxotrophy of CPSII-deficient mutants, restoring parasite growth and virulence, engineered mutations within, or proximal to, but the catalytic triad of the N-terminal glutamine amidotransferase domain inactivates the complementation activity of CPSII and demonstrates a critical dependence on the apicomplexan CPSII GATase domain in vivo
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pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
25 - 80
-
The activity of the CPS.A-CPS.B complex remains unchanged between 25-60 °C and then increases 54% at 70°C. The loss of catalytic activity above 80°C is indicated of thermal denaturation. The large increase in thermal stability at temperatures prior to thermal denaturation is not observed in the CPS.A-CPS.B-glutaminase-subunit complex.
additional information
-
the thermal stability of cloned enzyme is identical regardless of the growth temperature of Bacillus stearothermophilus between 42°C and 63°C. The thermal stability of the cloned enzyme is not affected by expression at 37°C in Bacillus subtilis or E. coli
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GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
enzyme is most stable in the presence of 30% dimethyl sulfoxide, 5% glycerol, 1 mM dithiothreitol. MgATP2-, MgUTP2-, and 5-phospho-alpha-D-ribosyl 1-diphosphate effectively stabilize
-
treatment with 1 M potassium thiocyanate results in reversible dissociation into its subunits which retain catalytic activity
-
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OXIDATION STABILITY
ORGANISM
UNIPROT
LITERATURE
Gln-dependent function of carbamoyl-phosphate synthase is inactivated by incubating the enzyme in air with low concentrations of dithiothreitol, glutathione, Cys, homocysteine, or 2-mercaptoethanol
-
1731
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-80°C, 30% dimethyl sulfoxide, 5% glycerol, 1 mM dithiothreitol, stable for 6 months
-
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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
multienzyme complex of carbamoyl-phosphate synthase(glutamine), aspartate carbamoyltransferase and dihydroorotase
-
multifunctional protein bearing carbamoyl-phosphate synthase, aspartate transcarbamoylase, and dihydroorotase activities
-
recombinant mutant enzymes from Escherichia coli strain RC50 to over 95% purity
-
recombinant wild-type and mutant enzymes
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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
carbamoyl-phosphate synthase belonging to the pyrimidine pathway
-
construction and expression of a functional minigene of CPSII for functional complementation of mutants, overview
-
Escherichia coli and chimeric enzyme are expressed in Escherichia coli L673
-
expression in Escherichia coli
expression of wild-type and mutant enzymes in Escherichia coli strain L673
-
gene carB, DNA and amino acid sequence determination and analysis
multifunctional protein, that catalyzes the first three steps in de novo pyrimidine biosynthesis, expression in Escherichia coli
-
subcloning of each of the subunits into the pET-28b vector, expression in Escherichia coli
-
the plasmids containing the carAB genes are transformed in the RC50 cell line of Escherichia coli for expression of the wild-type and mutant forms of CPS
-
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
accumulation of CPSase gene transcripts is highly reduced following seedling incubation with uridine
carB expression is partly downregulated by ammonium treatment, while carA expression is unaffected
-
mRNA levels of MtCPSs1 and MtCPSs2 are significantly increased after supplying plants with ornithine alone or in combination with uridine or arginine compared to plants treated with only uridine or arginine, respectively
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
drug development
-
CPSII is a potential parasite-selective target for drug development
medicine
-
serum samples from patients with acute liver failure due to acetaminophen, Wilson disease, or ischemia show readily detectable CPS1 that i not observed in patients with chronic viral hepatitis or in control donors. CPS1 rapidly decreases to undetectable levels in sera of patients with acetaminophen-related acute liver failure who ultimately recover
EXTERNAL LINKS (specific for EC-Number 6.3.5.5)
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Release 2026.1 (March 2026)
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