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Literature summary extracted from

  • Cockrell, G.M.; Kantrowitz, E.R.
    Metal ion involvement in the allosteric mechanism of Escherichia coli aspartate transcarbamoylase (2012), Biochemistry, 51, 7128-7137.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.1.3.2 overexpression in Escherichia coli strain EK1104 transformed with plasmid pEK15212 containing the Escherichia coli pyrBI gene Escherichia coli

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.1.3.2 purified recombinant ATCase in complex with UTP, CTP, or dCTP, dialysis of 20 mg/ml protein against 40 mM sodium citrate, 1 mM 2-mercaptoethanol, 0.2 mM EDTA, and 1.0 mM CTP, pH 5.7, at 20°C, 1 week, transfer of dialysis buttons to 2 mL of crystallization buffer with 5 mM UTP and 5 mM MgCl2 and equilibration for 12 h, and to crystallization buffer containing 5 mM UTP and 5 mM MgCl2 for 12 h, respectively, cryoprotection in 20% 2-methyl-2,4-pentanediol in UTP-Mg2+ crystallization buffer, X-ray diffraction structure determination and analysis at 1.9-2.1 A resolution Escherichia coli

Protein Variants

EC Number Protein Variants Comment Organism
2.1.3.2 D19A a regulatory mutant, which does not exhibit UTP synergistic inhibition Escherichia coli
2.1.3.2 H20A a regulatory mutant, which does not exhibit UTP synergistic inhibition Escherichia coli
2.1.3.2 K56A a regulatory mutant, which does not exhibit UTP synergistic inhibition Escherichia coli
2.1.3.2 K60A a regulatory mutant, which does not exhibit UTP synergistic inhibition Escherichia coli
2.1.3.2 K6A a regulatory mutant, which does not exhibit UTP synergistic inhibition Escherichia coli
2.1.3.2 L7A a regulatory mutant, which does not exhibit UTP synergistic inhibition Escherichia coli

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.1.3.2 CTP demetaled CTP, synergistic inhibition with UTP, while UTP alone has little or no influence on the enzyme activity, mechanism, overview. Binding of UTP can enhance the binding of CTP and presence of a metal ion such as Mg2+ is required for synergistic inhibition. Structure of the ATCase-CTP-UTP-Mg2+ complex Escherichia coli
2.1.3.2 dCTP
-
Escherichia coli
2.1.3.2 additional information conformational changes due to nucleotide binding, overview Escherichia coli
2.1.3.2 UTP inhibition with CTP, while UTP alone has little or no influence on the enzyme activity, mechanism, overview. UTP, in the presence of dCTP or CTP, binds at a site on a regulatory side chain that does not overlap the CTP/dCTP site, and the triphosphates of the two nucleotides are parallel to each other with a metal ion, in this case Mg2+, coordinated between the beta- and gamma-phosphates of the two nucleotides. UTP binds more tightly in the presence of CTP. Structure of the ATCase-CTP-UTP-Mg2+ complex Escherichia coli

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.1.3.2 additional information
-
additional information allosteric mechanism with metal ion involvement, overview Escherichia coli

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.1.3.2 Mg2+ required for synergistic inhibition of the enzyme by CTP and UTP, metal binding site in the allosteric regulatory site of ATCase, overview Escherichia coli

Natural Substrates/ Products (Substrates)

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

Organism

EC Number Organism UniProt Comment Textmining
2.1.3.2 Escherichia coli P0A786
-
-

Purification (Commentary)

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

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.1.3.2 L-aspartate + carbamoyl phosphate
-
Escherichia coli phosphate + N-carbamoyl-L-aspartate
-
?
2.1.3.2 additional information conformational changes due to nucleotide binding, overview Escherichia coli ?
-
?

Synonyms

EC Number Synonyms Comment Organism
2.1.3.2 aspartate transcarbamoylase
-
Escherichia coli
2.1.3.2 ATCase
-
Escherichia coli

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.1.3.2 25
-
assay at Escherichia coli

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.1.3.2 7
-
assay at Escherichia coli

General Information

EC Number General Information Comment Organism
2.1.3.2 metabolism aspartate transcarbamoylase allosterically regulates pyrimidine nucleotide biosynthesis Escherichia coli
2.1.3.2 physiological function aspartate transcarbamoylase allosterically regulates pyrimidine nucleotide biosynthesis Escherichia coli