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Crystallization and preliminary X‐ray study of Vibrio cholerae uridine phosphorylase in complex with 6‐methyluracil
Author(s) -
Prokofev Igor I.,
Lashkov Alexander A.,
Gabdulkhakov Azat G.,
Dontsova Mariya V.,
Seregina Tatyana A.,
Mironov Alexander S.,
Betzel Christian,
Mikhailov Al'bert M.
Publication year - 2014
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 37
ISSN - 2053-230X
DOI - 10.1107/s2053230x13031877
Subject(s) - vibrio cholerae , crystallization , nucleotide salvage , chemistry , microbiology and biotechnology , uridine , biochemistry , biology , bacteria , rna , nucleotide , organic chemistry , genetics , gene
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 ( Vch UPh), purification of this enzyme, crystallization of the complex of Vch UPh with 6‐MU, and X‐ray data collection and preliminary X‐ray analysis of the Vch UPh–6‐MU complex at atomic resolution are reported.