title: Crystallization and preliminary X-ray crystallographic studies of glutaredoxin 2 from Saccharomyces cerevisiae in different oxidation states.
abstract: 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 6xHis-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 A for untreated and oxidized Grx2, respectively, using a synchrotron-radiation source. The crystals belong to space group P4(1)2(1)2, with similar unit-cell parameters.
authors.affiliation: Departamento de Biologia, Instituto de Biociências, Universidade de São Paulo, Rua do Matão 277, São Paulo, SP, Brazil.
authors.forename: Karen Fulan
authors.initials: KF
authors.lastname: Discola
authors.forename: Marcos Antonio de
authors.initials: MA
authors.lastname: Oliveira
authors.forename: Gustavo Monteiro
authors.initials: GM
authors.lastname: Silva
authors.forename: José Antonio
authors.initials: JA
authors.lastname: Barcena
authors.forename: Pablo
authors.initials: P
authors.lastname: Porras
authors.forename: Alicia
authors.initials: A
authors.lastname: Padilla
authors.forename: Luis Eduardo Soares
authors.initials: LE
authors.lastname: Netto
authors.forename: Beatriz Gomes
authors.initials: BG
authors.lastname: Guimarães
mesh_terms: D003001:Cloning, Molecular; D003460:Crystallization / Q000379:methods; D004926:Escherichia coli / Q000235:genetics; D054477:Glutaredoxins; D006639:Histidine / Q000737:chemistry; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000737:chemistry*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry*; D014961:X-Ray Diffraction; D020122:tert-Butylhydroperoxide
chemical_list: D054477:Glutaredoxins; D029701:Saccharomyces cerevisiae Proteins; D006639:Histidine; D020122:tert-Butylhydroperoxide; D010088:Oxidoreductases