title: Crystallization and preliminary X-ray diffraction analysis of NADPH-dependent thioredoxin reductase I from Saccharomyces cerevisiae.
abstract: 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 His6-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 A using a synchrotron-radiation source. The crystal belongs to the centred monoclinic space group C2, with unit-cell parameters a = 127.97, b = 135.41, c = 75.81 A, beta = 89.95 degrees. The crystal structure was solved by molecular-replacement methods and structure refinement is in progress.
authors.affiliation: Departamento de Biologia, Instituto de Biociências, Universidade de São Paulo, Rua do Matão 277, São Paulo SP, Brazil.
authors.forename: Marcos Antonio de
authors.initials: MA
authors.lastname: Oliveira
authors.forename: Karen Fulan
authors.initials: KF
authors.lastname: Discola
authors.forename: Simone Vidigal
authors.initials: SV
authors.lastname: Alves
authors.forename: João Alexandre Ribeiro Gonçalves
authors.initials: JA
authors.lastname: Barbosa
authors.forename: Francisco Javier
authors.initials: FJ
authors.lastname: Medrano
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; D006639:Histidine; D011092:Polyethylene Glycols; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry*; D013880:Thioredoxin-Disulfide Reductase / Q000737:chemistry*; D014835:Volatilization; D014961:X-Ray Diffraction
chemical_list: D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D011092:Polyethylene Glycols; D006639:Histidine; D013880:Thioredoxin-Disulfide Reductase