In:
Acta Crystallographica Section F Structural Biology and Crystallization Communications, International Union of Crystallography (IUCr), Vol. 68, No. 11 ( 2012-11-01), p. 1394-1397
Abstract:
A high-resolution structure of the complex of Vibrio cholerae uridine phosphorylase ( Vch UPh) with its physiological ligand thymidine is important in order to determine the mechanism of the substrate specificity of the enzyme and for the rational design of pharmacological modulators. Here, the expression and purification of Vch UPh and the crystallization of its complex with thymidine are reported. Conditions for crystallization were determined with an automated Cartesian Dispensing System using The Classics, MbClass and MbClass II Suites crystallization kits. Crystals of the Vch UPh–thymidine complex (of dimensions ∼200–350 µm) were grown by the sitting-drop vapour-diffusion method in ∼7 d at 291 K. The crystallization solution consisted of 1.5 µl Vch UPh (15 mg ml −1 ), 1 µl 0.1 M thymidine and 1.5 µl reservoir solution [15%( w / v ) PEG 4000, 0.2 M MgCl 2 .6H 2 O in 0.1 M Tris–HCl pH 8.5]. The crystals diffracted to 2.12 Å resolution and belonged to space group P 2 1 (No. 4), with unit-cell parameters a = 91.80, b = 95.91, c = 91.89 Å, β = 119.96°. The Matthews coefficient was calculated as 2.18 Å 3 Da −1 ; the corresponding solvent content was 43.74%.
Type of Medium:
Online Resource
ISSN:
1744-3091
DOI:
10.1107/S1744309112041401
Language:
Unknown
Publisher:
International Union of Crystallography (IUCr)
Publication Date:
2012
detail.hit.zdb_id:
2175956-X
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