Purification, crystallization and preliminary crystallographic investigation of FrnE, a disulfide oxidoreductase from Deinococcus radiodurans.
In prokaryotes, Dsb proteins catalyze the formation of native disulfide bonds through an oxidative folding pathway and are part of the cell machinery that protects proteins from oxidative stress. Deinococcus radiodurans is an extremophile which shows unparalleled resistance to ionizing radiation and oxidative stress. It has a strong mechanism to protect its proteome from oxidative damage. The genome of Deinococcus shows the presence of FrnE, a Dsb protein homologue that potentially provides the bacterium with oxidative stress tolerance. Here, crystallization and preliminary X-ray crystallographic analysis of FrnE from D. radiodurans are reported. Diffraction-quality single crystals were obtained using the hanging-drop vapour-diffusion method with reservoir solution consisting of 100 mM sodium acetate pH 5.0, 10% PEG 8000, 15-20% glycerol. Diffraction data were collected on an Agilent SuperNova system using a microfocus sealed-tube X-ray source. The crystal diffracted to 1.8 Å resolution at 100 K. The space group of the crystal was found to be P2₁22₁, with unit-cell parameters a=47.91, b=62.94, c=86.75 Å, α=β=γ=90°. Based on Matthews coefficient analysis, one monomer per asymmetric unit is present in the crystal, with a solvent content of approximately 45%.
Solid State Physics Department, Bhabha Atomic Research Centre, Trombay, Mumbai 400 085, India.
Lata
L
Panicker
Molecular Biology Division, Bhabha Atomic Research Centre, Trombay, Mumbai 400 085, India.
Hari Sharan
HS
Misra
Solid State Physics Department, Bhabha Atomic Research Centre, Trombay, Mumbai 400 085, India.
Subhash Chandra
SC
Bihani
D001426:Bacterial Proteins / Q000737:chemistry* / Q000302:isolation & purification; D003460:Crystallization; D018360:Crystallography, X-Ray; D034301:Deinococcus / Q000201:enzymology*; D004220:Disulfides / Q000737:chemistry / Q000302:isolation & purification; D010088:Oxidoreductases / Q000737:chemistry* / Q000302:isolation & purification
D001426:Bacterial Proteins; D004220:Disulfides; D010088:Oxidoreductases
Deinococcus radiodurans; Dsb proteins; FrnE; disulfide oxidoreductase; oxidative stress