Literature summary for 2.1.3.2 extracted from

Harris, K.M.; Cockrell, G.M.; Puleo, D.E.; Kantrowitz, E.R.
Crystallographic snapshots of the complete catalytic cycle of the unregulated aspartate transcarbamoylase from Bacillus subtilis (2011), J. Mol. Biol., 411, 190-200.

Cloned(Commentary)

Cloned (Comment)	Organism
ATCase overexpression in Escherichia coli strain strain EK1104	Bacillus subtilis

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant enzyme alone or complex with substrate carbamoyl phosphate or inhibitor N-phosphonacetyl-L-aspartate, hanging drop vapor diffusion method, 12 mg/ml protein is mixed with an equal volume of crystallization buffer containing 1.7 M (NH4)2SO4, 0.1 M Tris-HCl, pH 8.5, and 2.0% PEG 200, and equilibratopn over a reservoir of 0.5 ml of crystallization buffer at 20°C, 1 week, for substrate bound enzyme the crystals are soaked in mother liquor containing the ligand at 13.3 mM, for inhibitor bound form, the enzyme is crystallized as described using crystallization buffer containing 0.1 M potassium bromide, 0.1 M N-cyclohexyl-3-aminopropanesulfonic acid, pH 10.0, and 18% PEG 8000, 20°C, 1 week, X-ray diffraction structure determination and analysis at 2.1-2.6 A resolution	Bacillus subtilis

Crystallization (Comment)

Organism

purified recombinant enzyme alone or complex with substrate carbamoyl phosphate or inhibitor N-phosphonacetyl-L-aspartate, hanging drop vapor diffusion method, 12 mg/ml protein is mixed with an equal volume of crystallization buffer containing 1.7 M (NH4)2SO4, 0.1 M Tris-HCl, pH 8.5, and 2.0% PEG 200, and equilibratopn over a reservoir of 0.5 ml of crystallization buffer at 20°C, 1 week, for substrate bound enzyme the crystals are soaked in mother liquor containing the ligand at 13.3 mM, for inhibitor bound form, the enzyme is crystallized as described using crystallization buffer containing 0.1 M potassium bromide, 0.1 M N-cyclohexyl-3-aminopropanesulfonic acid, pH 10.0, and 18% PEG 8000, 20°C, 1 week, X-ray diffraction structure determination and analysis at 2.1-2.6 A resolution

Bacillus subtilis

Inhibitors

Inhibitors	Comment	Organism	Structure
N-phosphonacetyl-L-aspartate	a bisubstrate/transition state analogue, binding structure, in silico docking and electrostatic calculations, overview	Bacillus subtilis

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
L-aspartate + carbamoyl phosphate	Bacillus subtilis	-	phosphate + N-carbamoyl-L-aspartate	-	?

Organism

Organism	UniProt	Comment	Textmining
Bacillus subtilis	P05654	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant enzyme from Escherichia coli strain EK114 by anion exchange chromatography, ammonium sulfate fractionation, and hydrophobic interaction chromatography	Bacillus subtilis

Reaction

Reaction	Comment	Organism	Reaction ID
carbamoyl phosphate + L-aspartate = phosphate + N-carbamoyl-L-aspartate	the catalytic cycle of ATCase leads from the ordered binding of the substrates to the formation and decomposition of the tetrahedral intermediate and to the ordered release of the products from the active site, ordered-binding mechanism, detailed overview	Bacillus subtilis	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
L-aspartate + carbamoyl phosphate	-	Bacillus subtilis	phosphate + N-carbamoyl-L-aspartate	-	?
L-aspartate + carbamoyl phosphate	carbamoyl phosphate binding structure, in silico docking and electrostatic calculations, overview	Bacillus subtilis	phosphate + N-carbamoyl-L-aspartate	-	?

Synonyms

Synonyms	Comment	Organism
aspartate transcarbamoylase	-	Bacillus subtilis
ATCase	-	Bacillus subtilis

General Information

General Information	Comment	Organism
malfunction	analysis of the conformational changes shows that there is a lack of cooperativity in trimeric ATCases that do not possess regulatory subunits	Bacillus subtilis
metabolism	ATCase catalyzes one of the first reactions in pyrimidine nucleotide biosynthesis	Bacillus subtilis