Expression, purification and preliminary crystallographic analysis of phosphoribosyl isomerase (PriA) from Streptomyces coelicolor

• Published in: Acta crystallographica. Section D, Biological crystallography. Biological crystallography., 2004-03, Vol.60 (3), p.534-536
• Main Authors: Wright, Helena, Barona-Gómez, Francisco, Hodgson, David A., Fülöp, Vilmos
• Format: Article
• Language: English
• Subjects: Aldose-Ketose Isomerases - chemistry; Aldose-Ketose Isomerases - genetics; Aldose-Ketose Isomerases - isolation & purification; Aldose-Ketose Isomerases - metabolism; Bacterial Proteins - chemistry; Bacterial Proteins - genetics; Bacterial Proteins - isolation & purification; Bacterial Proteins - metabolism; bifunctional enzyme; Crystallization; Crystallography, X-Ray; Gene Expression; Histidine - biosynthesis; Histidine - chemistry; histidine and tryptophan biosynthesis; phosphoribosyl isomerase; Recombinant Fusion Proteins - chemistry; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - isolation & purification; Recombinant Fusion Proteins - metabolism; Streptomyces - enzymology; Streptomyces - genetics; Tryptophan - biosynthesis
• ISSN: 1399-0047; 0907-4449; 1399-0047
• DOI: 10.1107/S0907444903028877

The priA gene encoding the enzyme phosphoribosyl isomerase from Streptomyces coelicolor, a novel bifunctional enzyme involved in both histidine and tryptophan biosynthesis, was heterologously expressed and purified in Escherichia coli as an N‐terminal His‐tag fusion. The purified recombinant enzyme was crystallized using the hanging‐drop method in 1.50 M ammonium sulfate and 100 mM sodium citrate pH 4.8. Crystals were obtained of up to 0.05 × 0.05 × 0.3 mm in size. A full data set to 2 Å resolution was collected at the ESRF beamline ID14‐1 and space group P31,221 was assigned, with unit‐cell parameters a = 65.1, c = 104.7 Å.