title: Crystallization and X-ray diffraction studies of cellobiose phosphorylase from Cellulomonas uda.
abstract: Disaccharide phosphorylases are able to catalyze both the synthesis and the breakdown of disaccharides and have thus emerged as attractive platforms for tailor-made sugar synthesis. Cellobiose phosphorylase from Cellulomonas uda (CPCuda) is an enzyme that belongs to glycoside hydrolase family 94 and catalyzes the reversible breakdown of cellobiose [beta-D-glucopyranosyl-(1,4)-D-glucopyranose] to alpha-D-glucose-1-phosphate and D-glucose. Crystals of ligand-free recombinant CPCuda and of its complexes with substrates and reaction products yielded complete X-ray diffraction data sets to high resolution using synchrotron radiation but suffered from significant variability in diffraction quality. In at least one case an intriguing space-group transition from a primitive monoclinic to a primitive orthorhombic lattice was observed during data collection. The structure of CPCuda was determined by maximum-likelihood molecular replacement, thus establishing a starting point for an investigation of the structural and mechanistic determinants of disaccharide phosphorylase activity.
authors.affiliation: Unit for Structural Biology and Biophysics, Laboratory for Protein Biochemistry and Biomolecular Engineering, K. L. Ledeganckstraat 35, Ghent University, B-9000 Ghent, Belgium.
authors.forename: Annelies
authors.initials: A
authors.lastname: Van Hoorebeke
authors.forename: Jan
authors.initials: J
authors.lastname: Stout
authors.forename: John
authors.initials: J
authors.lastname: Kyndt
authors.forename: Manu
authors.initials: M
authors.lastname: De Groeve
authors.forename: Ina
authors.initials: I
authors.lastname: Dix
authors.forename: Tom
authors.initials: T
authors.lastname: Desmet
authors.forename: Wim
authors.initials: W
authors.lastname: Soetaert
authors.forename: Jozef
authors.initials: J
authors.lastname: Van Beeumen
authors.forename: Savvas N
authors.initials: SN
authors.lastname: Savvides
mesh_terms: D040143:Cellulomonas / Q000201:enzymology*; D003460:Crystallization; D018360:Crystallography, X-Ray; D005964:Glucosyltransferases / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D008958:Models, Molecular; D020836:Protein Structure, Quaternary; D013379:Substrate Specificity
chemical_list: D005964:Glucosyltransferases; C034714:cellobiose phosphorylase