Crystallization and preliminary X-ray crystallographic studies of glutaredoxin 2 from Saccharomyces cerevisiae in different oxidation states
- Departamento de Biologia, Instituto de Biociências, Universidade de São Paulo, Rua do Matão 277, São Paulo, SP (Brazil)
- Departamento de Bioquímica y Biología Molecular, Campus de Rabanales, Edificio ‘Severo Ochoa’, Universidad de Córdoba, 14071 Córdoba (Spain)
Glutaredoxin 2 (Grx2) from S. cerevisiae was expressed, purified and crystallized using the hanging-drop vapour-diffusion method. Crystals were obtained from protein treated with t-butyl hydroperoxide and from a sample not submitted to pre-treatment. Both crystals belong to the tetragonal space group P4{sub 1}2{sub 1}2, with very similar unit-cell parameters, and diffraction data were collected to 2.05 and 2.15 Å resolution, respectively. Glutaredoxins are small (9–12 kDa) heat-stable proteins that are highly conserved throughout evolution; the glutaredoxin active site (Cys-Pro-Tyr-Cys) is conserved in most species. Five glutaredoxin genes have been identified in Saccharomyces cerevisiae; however, Grx2 is responsible for the majority of oxidoreductase activity in the cell, suggesting that its primary function may be the detoxification of mixed disulfides generated by reactive oxygen species (ROS). Recombinant Grx2 was expressed in Escherichia coli as a 6×His-tagged fusion protein and purified by nickel-affinity chromatography. Prior to crystallization trials, the enzyme was submitted to various treatments with reducing agents and peroxides. Crystals suitable for X-ray diffraction experiments were obtained from untreated protein and protein oxidized with t-butyl hydroperoxide (10 mM). Complete data sets were collected to resolutions 2.15 and 2.05 Å for untreated and oxidized Grx2, respectively, using a synchrotron-radiation source. The crystals belong to space group P4{sub 1}2{sub 1}2, with similar unit-cell parameters.
- OSTI ID:
- 22356104
- Journal Information:
- Acta Crystallographica. Section F, Vol. 61, Issue Pt 4; Other Information: PMCID: PMC1952414; PMID: 16511065; PUBLISHER-ID: fw5030; OAI: oai:pubmedcentral.nih.gov:1952414; Copyright (c) International Union of Crystallography 2005; Country of input: International Atomic Energy Agency (IAEA); ISSN 1744-3091
- Country of Publication:
- United Kingdom
- Language:
- English
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