Open AccessExpression, purification, crystallization and preliminary X‐ray diffraction analysis of human phosphoribosyl pyrophosphate synthetase 1 (PRS1)
Author(s) -
Tang Wenying,
Li Xiaowu,
Zhu Zhiqiang,
Tong Shuilong,
Li Xu,
Zhang Xiao,
Teng Maikun,
Niu Liwen
Publication year - 2006
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309106009067
Subject(s) - pyrophosphate , crystallization , ribose , purine metabolism , chemistry , crystallography , biosynthesis , molecule , phosphate , stereochemistry , biochemistry , enzyme , organic chemistry
Phosphoribosyl pyrophosphate synthetase (PRS; EC 2.7.6.1) catalyzes the reaction of ribose‐5‐phosphate (R5P) with ATP to yield AMP and PRPP (5‐phosphoribosyl‐1‐pyrophosphate), which is necessary for the de novo and salvage pathways of purine‐, pyrimidine‐ and pyridine‐nucleotide biosynthesis. PRPP is a metabolite that is required at all times in the cell and is thus central to life. In this study, human PRS1 was produced in Escherichia coli in soluble form and purified to homogeneity. Crystals in complex with Mg 2+ , inorganic phosphate (P i ) and ATP were obtained by the hanging‐drop vapour‐diffusion method. Diffraction data were collected to 2.6 Å resolution. The crystal belongs to space group R 3, with unit‐cell parameters a = b = 168.846, c = 61.857 Å, assuming two molecules in the asymmetric unit and a volume‐to‐weight ratio of 2.4 Å 3 Da −1 , which was consistent with the result calculated from the self‐rotation function.