Purification and crystallization of Kokobera virus helicase.
Kokobera virus is a mosquito-borne flavivirus belonging, like West Nile virus, to the Japanese encephalitis virus serocomplex. The flavivirus genus is characterized by a positive-sense single-stranded RNA genome. The unique open reading frame of the viral RNA is transcribed and translated as a single polyprotein which is post-translationally cleaved to yield three structural and seven nonstructural proteins, one of which is the NS3 gene that encodes a C-terminal helicase domain consisting of 431 amino acids. Helicase inhibitors are potential antiviral drugs as the helicase is essential to viral replication. Crystals of the Kokobera virus helicase domain were obtained by the hanging-drop vapour-diffusion method. The crystals belong to space group P3(1)21 (or P3(2)21), with unit-cell parameters a = 88.6, c = 138.6 A, and exhibit a diffraction limit of 2.3 A.
De Colibus
Luigi
L
Department of Genetics and Microbiology, University of Pavia, Via Ferrata 1, 27100 Pavia, Italy.
Speroni
Silvia
S
Coutard
Bruno
B
Forrester
Naomi L
NL
Gould
Ernest
E
Canard
Bruno
B
Mattevi
Andrea
A
D003460:Crystallization; D005416:Flavivirus / Q000201:enzymology*; D017434:Protein Structure, Tertiary; D020365:RNA Helicases / Q000737:chemistry* / Q000302:isolation & purification; D014764:Viral Proteins / Q000737:chemistry* / Q000302:isolation & purification
D014764:Viral Proteins; D020365:RNA Helicases