title: Cloning, expression, purification, crystallization and preliminary structure determination of glucose-1-phosphate uridylyltransferase (UgpG) from Sphingomonas elodea ATCC 31461 bound to glucose-1-phosphate.
abstract: The cloning, expression, purification, crystallization and preliminary crystallographic analysis of glucose-1-phosphate uridylyltransferase (UgpG) from Sphingomonas elodea ATCC 31461 bound to glucose-1-phosphate are reported. Diffraction data sets were obtained from seven crystal forms in five different space groups, with highest resolutions ranging from 4.20 to 2.65 A. The phase problem was solved for a P2(1) crystal form using multiple isomorphous replacement with anomalous scattering from an osmium derivative and a SeMet derivative. The best native crystal in space group P2(1) has unit-cell parameters a = 105.5, b = 85.7, c = 151.8 A, beta = 105.2 degrees . Model building and refinement are currently under way.
authors.affiliation: European Synchrotron Radiation Facility, BP 220, F-38043 Grenoble CEDEX, France.
authors.forename: D
authors.initials: D
authors.lastname: Aragão
authors.forename: A R
authors.initials: AR
authors.lastname: Marques
authors.forename: C
authors.initials: C
authors.lastname: Frazão
authors.forename: F J
authors.initials: FJ
authors.lastname: Enguita
authors.forename: M A
authors.initials: MA
authors.lastname: Carrondo
authors.forename: A M
authors.initials: AM
authors.lastname: Fialho
authors.forename: I
authors.initials: I
authors.lastname: Sá-Correia
authors.forename: E P
authors.initials: EP
authors.lastname: Mitchell
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D001665:Binding Sites / Q000235:genetics; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray / Q000379:methods; D015964:Gene Expression Regulation, Bacterial; D005958:Glucosephosphates / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D020578:Sphingomonas / Q000201:enzymology* / Q000235:genetics*; D013379:Substrate Specificity / Q000235:genetics; D005957:UTP-Glucose-1-Phosphate Uridylyltransferase / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D005958:Glucosephosphates; C031590:glucose-1-phosphate; D005957:UTP-Glucose-1-Phosphate Uridylyltransferase