Literature summary for 2.1.3.2 extracted from

Guo, W.; West, J.M.; Dutton, A.S.; Tsuruta, H.; Kantrowitz, E.R.
Trapping and structure determination of an intermediate in the allosteric transition of aspartate transcarbamoylase (2012), Proc. Natl. Acad. Sci. USA, 109, 7741-7746.

Cloned(Commentary)

Cloned (Comment)	Organism
expression of wild-type and mutant enzymes in Escherichia coli strain EK1104 containing the plasmid pEK695	Escherichia coli

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant mutant enzyme K164E/E239K, mixing of 10 mg/ml enzyme in 50 mM Tris-acetate, pH 8.3, with 0.002 ml of crystallization buffer containing 16% w/v PEG 4000, 0.04 M Na2MoO4-2H2O, 0.04 M N-cyclohexyl-3-aminopropanesulfonic acid, and 30 mM Tris-acetate, pH 8.75, and equilibration over a reservoir of crystallization buffer of 1.0 ml, 20°C, 2 weeks, X-ray diffraction structure determination and analysis	Escherichia coli

Crystallization (Comment)

Organism

purified recombinant mutant enzyme K164E/E239K, mixing of 10 mg/ml enzyme in 50 mM Tris-acetate, pH 8.3, with 0.002 ml of crystallization buffer containing 16% w/v PEG 4000, 0.04 M Na2MoO4-2H2O, 0.04 M N-cyclohexyl-3-aminopropanesulfonic acid, and 30 mM Tris-acetate, pH 8.75, and equilibration over a reservoir of crystallization buffer of 1.0 ml, 20°C, 2 weeks, X-ray diffraction structure determination and analysis

Escherichia coli

Protein Variants

Protein Variants	Comment	Organism
D236A	site-directed mutagenesis, the wild-type enzyme shows o change in structure by SAXS through the temperature range of 4°C to 55°C, whereas the D236A ATCase exhibits a large shift toward the T state between 4°C and 30°C, with a minor shift back toward the R state between 30°C and 45°C	Escherichia coli
K164E/E239K	site-directed mutagenesis, a mutant aspartate transcarbamoylase exists in an intermediate quaternary structure between the canonical T and R structures, crystal structure and quaternary conformation analysis, detailed overview. pH-Dependent structural alteration consistent with either a pH-induced conformational change or a pH-induced alteration in the T to R equilibrium	Escherichia coli

Natural Substrates/ Products (Substrates)

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

Organism

Organism	UniProt	Comment	Textmining
Escherichia coli	P0A786	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant wild-type and mutant enzymes from Escherichia coli strain EK1104 by isoelectric precipitation, anion exchange, and hydrophobic interaction chromatography, and gel filtration	Escherichia coli

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
L-aspartate + carbamoyl phosphate	-	Escherichia coli	phosphate + N-carbamoyl-L-aspartate	-	?

Synonyms

Synonyms	Comment	Organism
aspartate transcarbamoylase	-	Escherichia coli

Temperature Optimum [°C]

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

Temperature Range [°C]

Temperature Minimum [°C]	Temperature Maximum [°C]	Comment	Organism
additional information	-	the wild-type enzyme shows o change in structure by SAXS through the temperature range of 4°C to 55°C, whereas the D236A ATCase exhibits a large shift toward the T state between 4°C and 30°C, with a minor shift back toward the R state between 30°C and 45°C	Escherichia coli

pH Optimum

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

General Information

General Information	Comment	Organism
additional information	allosteric transition between the T and R enzyme states	Escherichia coli