Expression and x-ray structural determination of the nucleoprotein of lassa fever virus

Xiaoxuan Qi, Wenjian Wang, Haohao Dong, Yuying Liang, Changjiang Dong, Hinh Ly

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

We describe methods to express the nucleoprotein (NP) of Lassa fever virus (LASV) in E. coli, to purify and crystallize it using the sitting-drop vapor diffusion method. The crystals were screened using Rigaku micro-007 X-ray generator and a dataset was collected at a resolution of 2.36 Å. The crystals belong to space group P3, with the unit cell parameters a = b = 176.35 Å, c = 56.40 Å, α = β = 90°, and γ = 120°. Using the X-ray diffraction method, we constructed a three-dimensional structure of the LASV NP that should aid in the development of novel therapeutic strategies against this virus, for which vaccine and effective treatment modalities are currently unavailable.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages179-188
Number of pages10
DOIs
StatePublished - 2018

Publication series

NameMethods in Molecular Biology
Volume1604
ISSN (Print)1064-3745

Bibliographical note

Funding Information:
We thank Dr. Louise Major for providing the pLou3 plasmid, Dr. Catherine Botting for assistance with mass spectroscopy, Dr. Zaitsev Slava for helping with data collection, and Dr. Bjoern Meyer for prompt editorial assistance. This work was supported in part by the NIAID/NIH through the new-direction awards mechanism of the SERCEB grant (U54-AI057157) to YL and HL, by the NIAID/NIH R01 AI083409 to YL, R01 AI093580 and R56 AI091805 to HL, and the Medical Research Council grant (G1100110/1) to CJD.

Keywords

  • Arenavirus
  • Large-scale expression
  • Lassa virus
  • Nucleoprotein
  • X-ray structure

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