title: Crystallization and preliminary X-ray study of Vibrio cholerae uridine phosphorylase in complex with 6-methyluracil.
abstract: Uridine phosphorylase catalyzes the phosphorolysis of ribonucleosides, with the nitrogenous base and ribose 1-phosphate as products. Additionally, it catalyzes the reverse reaction of the synthesis of ribonucleosides from ribose 1-phosphate and a nitrogenous base. However, the enzyme does not catalyze the synthesis of nucleosides when the substrate is a nitrogenous base substituted at the 6-position, such as 6-methyluracil (6-MU). In order to explain this fact, it is essential to investigate the three-dimensional structure of the complex of 6-MU with uridine phosphorylase. 6-MU is a pharmaceutical agent that improves tissue nutrition and enhances cell regeneration by normalization of nucleotide exchange in humans. 6-MU is used for the treatment of diseases of the gastrointestinal tract, including infectious diseases. Here, procedures to obtain the uridine phosphorylase from the pathogenic bacterium Vibrio cholerae (VchUPh), purification of this enzyme, crystallization of the complex of VchUPh with 6-MU, and X-ray data collection and preliminary X-ray analysis of the VchUPh-6-MU complex at atomic resolution are reported.
authors.affiliation: A. V. Shubnikov Institute of Crystallography, Russian Academy of Sciences, Leninsky Prospect 59, Moscow 117333, Russian Federation.
authors.forename: Igor I
authors.initials: II
authors.lastname: Prokofev
authors.affiliation: A. V. Shubnikov Institute of Crystallography, Russian Academy of Sciences, Leninsky Prospect 59, Moscow 117333, Russian Federation.
authors.forename: Alexander A
authors.initials: AA
authors.lastname: Lashkov
authors.affiliation: A. V. Shubnikov Institute of Crystallography, Russian Academy of Sciences, Leninsky Prospect 59, Moscow 117333, Russian Federation.
authors.forename: Azat G
authors.initials: AG
authors.lastname: Gabdulkhakov
authors.affiliation: A. V. Shubnikov Institute of Crystallography, Russian Academy of Sciences, Leninsky Prospect 59, Moscow 117333, Russian Federation.
authors.forename: Mariya V
authors.initials: MV
authors.lastname: Dontsova
authors.affiliation: A. V. Shubnikov Institute of Crystallography, Russian Academy of Sciences, Leninsky Prospect 59, Moscow 117333, Russian Federation.
authors.forename: Tatyana A
authors.initials: TA
authors.lastname: Seregina
authors.affiliation: State Research Institute of Genetics and Selection of Industrial Microorganisms, 1st Dorozhny Proezd 1, Moscow 117545, Russian Federation.
authors.forename: Alexander S
authors.initials: AS
authors.lastname: Mironov
authors.affiliation: University of Hamburg, Notkestrasse 85, 22603 Hamburg, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Betzel
authors.affiliation: A. V. Shubnikov Institute of Crystallography, Russian Academy of Sciences, Leninsky Prospect 59, Moscow 117333, Russian Federation.
authors.forename: Al'bert M
authors.initials: AM
authors.lastname: Mikhailov
mesh_terms: D001665:Binding Sites; D055162:Biocatalysis; D003460:Crystallization; D018360:Crystallography, X-Ray; D008958:Models, Molecular; D014498:Uracil / Q000031:analogs & derivatives* / Q000737:chemistry; D014543:Uridine Phosphorylase / Q000737:chemistry*; D014734:Vibrio cholerae / Q000201:enzymology*
chemical_list: D014498:Uracil; C008378:6-methyluracil; D014543:Uridine Phosphorylase
keywords: 6-methyluracil; Vibrio cholerae; bacterial uridine phosphorylase