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Literature summary extracted from

  • Brugarolas, P.; Duguid, E.M.; Zhang, W.; Poor, C.B.; He, C.
    Structural and biochemical characterization of N5-carboxyaminoimidazole ribonucleotide synthetase and N5-carboxyaminoimidazole ribonucleotide mutase from Staphylococcus aureus (2011), Acta Crystallogr. Sect. D, 67, 707-715.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
5.4.99.18 gene purE, expression of His-tagged enzyme in Escherichia coli strain BL21 Star (DE3) Staphylococcus aureus
6.3.4.18 expressed in Escherichia coli BL21 Star (DE3) cells Staphylococcus aureus

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
5.4.99.18 purified recombinant detagged enzyme, crystallization at room temperature, 10 mg/ml protein is mixed with a reservoir solution consisting of 0.1 M sodium acetate trihydrate, pH 4.5, and 2 M ammonium sulfate, 24 h, X-ray diffraction structure determination and analysis at 1.45 Aresolution, molecular replacement using the structure of Bacillus anthracis PurE, PDB ID 1xmp, as template Staphylococcus aureus
6.3.4.18 hanging drop vapor diffusion method, using 0.1 M bis-Tris pH 6.5, 28% (w/v) polyethylene glycol monomethyl ether 2000 Staphylococcus aureus

Protein Variants

EC Number Protein Variants Comment Organism
5.4.99.18 H38F site-directed mutagenesis of the catalytic residue renders the enzyme inactive Staphylococcus aureus
5.4.99.18 H38N site-directed mutagenesis of the catalytic residue renders the enzyme inactive Staphylococcus aureus
5.4.99.18 H38W site-directed mutagenesis of the catalytic residue renders the enzyme inactive Staphylococcus aureus
6.3.4.18 F78I the mutant shows strongly reduced catalytic efficiency compared to the wild type enzyme Staphylococcus aureus
6.3.4.18 F78W the mutant shows strongly reduced catalytic efficiency compared to the wild type enzyme Staphylococcus aureus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
6.3.4.18 0.0139
-
5-amino-1-(5-phospho-D-ribosyl)imidazole wild type enzyme, in 50 mM HEPES, 80 mM KCl, 20 mM MgCl2, at 22°C, pH not specified in the publication Staphylococcus aureus
6.3.4.18 0.0432
-
ATP wild type enzyme, in 50 mM HEPES, 80 mM KCl, 20 mM MgCl2, at 22°C, pH not specified in the publication Staphylococcus aureus
6.3.4.18 0.0513
-
5-amino-1-(5-phospho-D-ribosyl)imidazole mutant enzyme F78I, in 50 mM HEPES, 80 mM KCl, 20 mM MgCl2, at 22°C, pH not specified in the publication Staphylococcus aureus
6.3.4.18 0.103
-
5-amino-1-(5-phospho-D-ribosyl)imidazole mutant enzyme F78W, in 50 mM HEPES, 80 mM KCl, 20 mM MgCl2, at 22°C, pH not specified in the publication Staphylococcus aureus
6.3.4.18 18.8
-
HCO3- wild type enzyme, in 50 mM HEPES, 80 mM KCl, 20 mM MgCl2, at 22°C, pH not specified in the publication Staphylococcus aureus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
5.4.99.18 Mg2+ required Staphylococcus aureus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
5.4.99.18 5-carboxyamino-1-(5-phospho-D-ribosyl)imidazole Staphylococcus aureus
-
5-amino-1-(5-phospho-D-ribosyl)imidazole-4-carboxylate
-
?
6.3.4.18 ATP + 5-amino-1-(5-phospho-D-ribosyl)imidazole + HCO3- + H+ Staphylococcus aureus
-
ADP + phosphate + 5-carboxyamino-1-(5-phospho-D-ribosyl)imidazole
-
?
6.3.4.18 ATP + 5-amino-1-(5-phospho-D-ribosyl)imidazole + HCO3- + H+ Staphylococcus aureus Newman
-
ADP + phosphate + 5-carboxyamino-1-(5-phospho-D-ribosyl)imidazole
-
?

Organism

EC Number Organism UniProt Comment Textmining
5.4.99.18 Staphylococcus aureus A0A0H3K7Y1 gene purE
-
6.3.4.18 Staphylococcus aureus A0A0H3KFM7
-
-
6.3.4.18 Staphylococcus aureus Newman A0A0H3KFM7
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
5.4.99.18 recombinant His-tagged enzyme from Escherichia coli strain BL21 Star (DE3) by nickel affinity chromatography, tag cleavage by TEV protease, and gel filtration Staphylococcus aureus
6.3.4.18 Ni-NTA column chromatography and Superdex 200 gel filtration Staphylococcus aureus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5.4.99.18 5-carboxyamino-1-(5-phospho-D-ribosyl)imidazole
-
Staphylococcus aureus 5-amino-1-(5-phospho-D-ribosyl)imidazole-4-carboxylate
-
?
6.3.4.18 ATP + 5-amino-1-(5-phospho-D-ribosyl)imidazole + HCO3- + H+
-
Staphylococcus aureus ADP + phosphate + 5-carboxyamino-1-(5-phospho-D-ribosyl)imidazole
-
?
6.3.4.18 ATP + 5-amino-1-(5-phospho-D-ribosyl)imidazole + HCO3- + H+
-
Staphylococcus aureus Newman ADP + phosphate + 5-carboxyamino-1-(5-phospho-D-ribosyl)imidazole
-
?

Subunits

EC Number Subunits Comment Organism
6.3.4.18 homodimer x-ray crystallography Staphylococcus aureus

Synonyms

EC Number Synonyms Comment Organism
5.4.99.18 N5-carboxyaminoimidazole ribonucleotide synthetase
-
Staphylococcus aureus
5.4.99.18 PurE
-
Staphylococcus aureus
6.3.4.18 N5-CAIR synthetase
-
Staphylococcus aureus
6.3.4.18 N5-carboxyaminoimidazole ribonucleotide synthetase
-
Staphylococcus aureus
6.3.4.18 PurK
-
Staphylococcus aureus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
5.4.99.18 30
-
assay at Staphylococcus aureus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
6.3.4.18 15.83
-
5-amino-1-(5-phospho-D-ribosyl)imidazole mutant enzyme F78I, in 50 mM HEPES, 80 mM KCl, 20 mM MgCl2, at 22°C, pH not specified in the publication Staphylococcus aureus
6.3.4.18 18
-
5-amino-1-(5-phospho-D-ribosyl)imidazole mutant enzyme F78W, in 50 mM HEPES, 80 mM KCl, 20 mM MgCl2, at 22°C, pH not specified in the publication Staphylococcus aureus
6.3.4.18 24.83
-
5-amino-1-(5-phospho-D-ribosyl)imidazole wild type enzyme, in 50 mM HEPES, 80 mM KCl, 20 mM MgCl2, at 22°C, pH not specified in the publication Staphylococcus aureus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
5.4.99.18 7.8
-
assay at Staphylococcus aureus

Cofactor

EC Number Cofactor Comment Organism Structure
6.3.4.18 ATP
-
Staphylococcus aureus

General Information

EC Number General Information Comment Organism
5.4.99.18 additional information His38 is essential for function, role of His38 as a general acid/base catalyst, structure analysis, comparison to Homo sapiens class II PurE enzyme, active site structure, overview Staphylococcus aureus

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
6.3.4.18 175
-
5-amino-1-(5-phospho-D-ribosyl)imidazole mutant enzyme F78W, in 50 mM HEPES, 80 mM KCl, 20 mM MgCl2, at 22°C, pH not specified in the publication Staphylococcus aureus
6.3.4.18 308
-
5-amino-1-(5-phospho-D-ribosyl)imidazole mutant enzyme F78I, in 50 mM HEPES, 80 mM KCl, 20 mM MgCl2, at 22°C, pH not specified in the publication Staphylococcus aureus
6.3.4.18 1783
-
5-amino-1-(5-phospho-D-ribosyl)imidazole wild type enzyme, in 50 mM HEPES, 80 mM KCl, 20 mM MgCl2, at 22°C, pH not specified in the publication Staphylococcus aureus