Expression, purification, crystallization and preliminary X-ray diffraction analysis of human phosphoribosyl pyrophosphate synthetase 1 (PRS1)
- Hefei National Laboratory for Physical Sciences at Microscale and School of Life Sciences, University of Science and Technology of China, 96 Jinzhai Road, Hefei, Anhui 230027 (China)
Crystals of human PRPP synthetase 1 (PRS1) in complex with Mg{sup 2+}, P{sub i} and ATP were obtained and diffraction data were collected to 2.6 Å resolution. 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{sup 2+}, inorganic phosphate (P{sub 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 R3, 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{sup 3} Da{sup −1}, which was consistent with the result calculated from the self-rotation function.
- OSTI ID:
- 22356325
- Journal Information:
- Acta Crystallographica. Section F, Vol. 62, Issue Pt 5; Other Information: PMCID: PMC2219982; PMID: 16682768; PUBLISHER-ID: pu5132; OAI: oai:pubmedcentral.nih.gov:2219982; Copyright (c) International Union of Crystallography 2006; Country of input: International Atomic Energy Agency (IAEA); ISSN 1744-3091
- Country of Publication:
- United Kingdom
- Language:
- English
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