Crystallization and preliminary X-ray crystallographic studies of glutaredoxin 2 from Saccharomyces cerevisiae in different oxidation states

摘要: Glutaredoxins are small (9–12 kDa) heat-stable proteins that highly conserved throughout evolution; the glutaredoxin active site (Cys-Pro-Tyr-Cys) is in most species. Five genes have been identified Saccharomyces cerevisiae; however, Grx2 responsible for majority of oxidoreductase activity cell, suggesting its primary function may be detoxification mixed disulfides generated by reactive oxygen species (ROS). Recombinant was expressed Escherichia coli as a 6×His-tagged fusion protein and purified nickel-affinity chromatography. Prior to crystallization trials, enzyme submitted various treatments with reducing agents peroxides. Crystals suitable X-ray diffraction experiments were obtained from untreated oxidized t-butyl hydroperoxide (10 mM). Complete data sets collected resolutions 2.15 2.05 A Grx2, respectively, using synchrotron-radiation source. The crystals belong space group P41212, similar unit-cell parameters.