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Literature summary for 5.4.99.18 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)

Cloned (Comment) Organism
gene purE, expression of His-tagged enzyme in Escherichia coli strain BL21 Star (DE3) Staphylococcus aureus

Crystallization (Commentary)

Crystallization (Comment) Organism
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

Protein Variants

Protein Variants Comment Organism
H38F site-directed mutagenesis of the catalytic residue renders the enzyme inactive Staphylococcus aureus
H38N site-directed mutagenesis of the catalytic residue renders the enzyme inactive Staphylococcus aureus
H38W site-directed mutagenesis of the catalytic residue renders the enzyme inactive Staphylococcus aureus

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required Staphylococcus aureus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
5-carboxyamino-1-(5-phospho-D-ribosyl)imidazole Staphylococcus aureus
-
5-amino-1-(5-phospho-D-ribosyl)imidazole-4-carboxylate
-
?

Organism

Organism UniProt Comment Textmining
Staphylococcus aureus A0A0H3K7Y1 gene purE
-

Purification (Commentary)

Purification (Comment) Organism
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

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5-carboxyamino-1-(5-phospho-D-ribosyl)imidazole
-
Staphylococcus aureus 5-amino-1-(5-phospho-D-ribosyl)imidazole-4-carboxylate
-
?

Synonyms

Synonyms Comment Organism
N5-carboxyaminoimidazole ribonucleotide synthetase
-
Staphylococcus aureus
PurE
-
Staphylococcus aureus

Temperature Optimum [°C]

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

pH Optimum

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

General Information

General Information Comment Organism
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