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

Journal Article (Journal Article)

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.

Full Text

Duke Authors

Cited Authors

  • Discola, KF; Oliveira, MAD; Silva, GM; Barcena, JA; Porras, P; Padilla, A; Netto, LES; Guimarães, BG

Published Date

  • April 2005

Published In

Volume / Issue

  • 61 / Pt 4

Start / End Page

  • 445 - 447

PubMed ID

  • 16511065

Pubmed Central ID

  • PMC1952414

Electronic International Standard Serial Number (EISSN)

  • 1744-3091

Digital Object Identifier (DOI)

  • 10.1107/s1744309105008730


  • eng