Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2007 Mar 1;63(Pt 3):193-5.
doi: 10.1107/S1744309107005283. Epub 2007 Feb 10.

Purification and crystallization of Kokobera virus helicase

Luigi De Colibus  1 , Silvia Speroni, Bruno Coutard, Naomi L Forrester, Ernest Gould, Bruno Canard, Andrea Mattevi
Affiliations

Purification and crystallization of Kokobera virus helicase

Luigi De Colibus et al. Acta Crystallogr Sect F Struct Biol Cryst Commun. .

Abstract

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.

PubMed Disclaimer

Figures

Figure 1
Figure 1
Representative protein samples from each stage of the purification analysed by SDS–PAGE [10%(w/v) acrylamide]. Lanes A and B correspond to the protein eluted from the nickel-affinity chromatography column and the protein eluted from final gel-filtration step, respectively. Lane M contains molecular-weight markers (kDa).
Figure 2
Figure 2
A typical crystal of KOKV helicase domain grown in a hanging-drop vapour-diffusion experiment.
Figure 3
Figure 3
Diffraction image of a KOKV helicase crystal collected on beamline ID23-EH1 at the ESRF. The resolution limits of the circles are in Å.
See this image and copyright information in PMC

References

    1. Abergel, C., Coutard, B., Byrne, D., Chenivesse, S., Claude, J. B., Deregnaucourt, C., Fricaux, T., Gianesini-Boutreux, C., Jeudy, S., Lebrun, R., Maza, C., Notredame, C., Poirot, O., Suhre, K., Varagnol, M. & Claverie, J. M. (2003). J. Struct. Funct. Genomics, 4, 141–157. - PubMed
    1. Benarroch, D., Selisko, B., Locatelli, G., Maga, G., Romette, J. L. & Canard, B. (2004). Virology, 328, 208–218. - PubMed
    1. Berrow, N. S., Bussow, K., Coutard, B., Diprose, J., Ekberg, M., Folkers, G. E., Levy, N., Lieu, V., Owens, R. J., Peleg, Y., Pinaglia, C., Quevillon-Cheruel, S., Salim, L., Scheich, C., Vincentelli, R. & Busso, D. (2006). Acta Cryst. D62, 1218–1226. - PubMed
    1. Collaborative Computational Project, Number 4 (1994). Acta Cryst. D50, 760–­763. - PubMed
    1. Doherty, R. L., Carley, J. G. & Gorman, B. M. (1964). Aust. J. Exp. Biol. Med. Sci.42, 149–164. - PubMed

Publication types

MeSH terms