Crystallization and preliminary X-ray diffraction analysis of protein 14 from Sulfolobus islandicus filamentous virus (SIFV).
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 P6(2)22 or P6(4)22, with unit-cell parameters a = b = 68.1, c = 132.4 A. Diffraction data were collected to a maximum acceptable resolution of 2.95 and 3.20 A for the SeMet-labelled and native protein, respectively.
Goulet
Adeline
A
Architecture et Fonction des Macromolécules Biologiques, CNRS and Universités d'Aix-Marseille I et II, UMR 6098, Case 932, 163 Avenue de Luminy, 13288 Marseille CEDEX 9, France.
Spinelli
Silvia
S
Campanacci
Valérie
V
Porciero
Sophie
S
Blangy
Stéphanie
S
Garrett
Roger A
RA
van Tilbeurgh
Herman
H
Leulliot
Nicolas
N
Basta
Tamara
T
Prangishvili
David
D
Cambillau
Christian
C
D003001:Cloning, Molecular; D003460:Crystallization; D023643:Lipothrixviridae / Q000737:chemistry*; D016944:Sulfolobus / Q000235:genetics / Q000821:virology*; D014764:Viral Proteins / Q000737:chemistry* / Q000235:genetics; D014961:X-Ray Diffraction / Q000379:methods
D014764:Viral Proteins