Open AccessCrystallization and preliminary X‐ray diffraction analysis of protein 14 from Sulfolobus islandicus filamentous virus (SIFV)
Author(s) -
Goulet Adeline,
Spinelli Silvia,
Campanacci Valérie,
Porciero Sophie,
Blangy Stéphanie,
Garrett Roger A.,
Van Tilbeurgh Herman,
Leulliot Nicolas,
Basta Tamara,
Prangishvili David,
Cambillau Christian
Publication year - 2006
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309106029150
Subject(s) - sulfolobus , crystallization , archaea , escherichia coli , crystallography , protein crystallization , biology , virus , x ray crystallography , chemistry , biochemistry , microbiology and biotechnology , diffraction , virology , gene , physics , organic chemistry , optics
A large‐scale programme has been embarked upon aiming towards the structural determination of conserved proteins from viruses infecting hyperthermophilic archaea. Here, the crystallization of protein 14 from the archaeal virus SIFV is reported. This protein, which contains 111 residues (MW 13 465 Da), was cloned and expressed in Escherichia coli with an N‐terminal His 6 tag and purified to homogeneity. The tag was subsequently cleaved and the protein was crystallized using PEG 1000 or PEG 4000 as a precipitant. Large crystals were obtained of the native and the selenomethionine‐labelled protein using sitting drops of 100–300 nl. Crystals belong to space group P 6 2 22 or P 6 4 22, with unit‐cell parameters a = b = 68.1, c = 132.4 Å. Diffraction data were collected to a maximum acceptable resolution of 2.95 and 3.20 Å for the SeMet‐labelled and native protein, respectively.