Crystallization and Preliminary X-Ray Diffraction Studies of N-Acyl-D-glucosamine 2-Epimerase from Porcine Kidney

• Published in: Journal of biochemistry (Tokyo) Vol. 120; no. 3; pp. 481 - 482
• Main Authors: Maru, Isafumi, Ohnish, Jun, Ohta, Yasuhiro, Hashimoto, Wataru, Tsukada, Yoji, Kousaku, Murata, Mikami, Bunzo
• Format: Journal Article
• Language: English
• Published: England Oxford University Press 01.09.1996
• Subjects: Animals; Carbohydrate Epimerases - biosynthesis; Carbohydrate Epimerases - chemistry; Carbohydrate Epimerases - isolation & purification; Carrier Proteins - biosynthesis; Carrier Proteins - chemistry; Carrier Proteins - isolation & purification; Cloning, Molecular; Crystallization; Crystallography, X-Ray; Escherichia coli; Kidney - enzymology; N-acyl-D-glucosamine 2-epimerase; porcine kidney; Recombinant Proteins - biosynthesis; Recombinant Proteins - chemistry; Recombinant Proteins - isolation & purification; reninbinding protein; Swine; X-ray crystallography
• ISSN: 0021-924X
• DOI: 10.1093/oxfordjournals.jbchem.a021437

N-Acyl-D-glucosamine 2-epimerase from porcine kidney, which was cloned and expressed in Escherichia coli, was crystallized by the vapor-diffusion method, using polyethylene glycol and ammonium acetate as precipitants. The crystals were resistant to X-ray radiation damage and diffracted to more than 2.0 Å resolution. The diffraction pattern indicated that the crystals belong to the orthorhombic system, space group P21,21 21 with unit-cell dimensions of a=78.1, b=97.2, and c=100.7 Å. It is supposed that the asymmetric unit consists of two N-acyl-D-glucosamine 2-epimerase molecules. Collection of data on the native crystals indicated that they are suitable for X-ray structural analysis.