Open AccessCrystallization and preliminary X‐ray diffraction analysis of NADPH‐dependent thioredoxin reductase I from Saccharomyces cerevisiae
Author(s) -
Discola Karen Fulan,
Alves Simone Vidigal,
De Oliveira Marcos Antonio,
Barbosa João Alexandre Ribeiro Gonçalves,
Medrano Francisco Javier,
Netto Luis Eduardo Soares,
Guimarães Beatriz Gomes
Publication year - 2005
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s174430910500758x
Subject(s) - saccharomyces cerevisiae , thioredoxin reductase , crystallization , chemistry , biochemistry , crystallography , materials science , thioredoxin , enzyme , yeast , organic chemistry
Thioredoxin reductase 1 (Trr1) from Saccharomyces cerevisiae is a member of the family of pyridine nucleotide‐disulfide oxidoreductases capable of reducing the redox‐active disulfide bond of the cytosolic thioredoxin 1 (Trx1) and thioredoxin 2 (Trx2). NADPH, Trr1 and Trx1 (or Trx2) comprise the thioredoxin system, which is involved in several biological processes, including the reduction of disulfide bonds and response to oxidative stress. Recombinant Trr1 was expressed in Escherichia coli as a His 6 ‐tagged fusion protein and purified by nickel‐affinity chromatography. The protein was crystallized using the hanging‐drop vapour‐diffusion method in the presence of PEG 3000 as precipitant after treatment with hydrogen peroxide. X‐ray diffraction data were collected to a maximum resolution of 2.4 Å using a synchrotron‐radiation source. The crystal belongs to the centred monoclinic space group C 2, with unit‐cell parameters a = 127.97, b = 135.41, c = 75.81 Å, β = 89.95°. The crystal structure was solved by molecular‐replacement methods and structure refinement is in progress.