Expression, purification and crystallization of a lyssavirus matrix (M) protein

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2008 Apr 1;64(Pt 4):258-62. doi: 10.1107/S1744309108004557. Epub 2008 Mar 21.

Abstract

The matrix (M) proteins of lyssaviruses (family Rhabdoviridae) are crucial to viral morphogenesis as well as in modulating replication and transcription of the viral genome. To date, no high-resolution structural information has been obtained for full-length rhabdovirus M. Here, the cloning, expression and purification of the matrix proteins from three lyssaviruses, Lagos bat virus (LAG), Mokola virus and Thailand dog virus, are described. Crystals have been obtained for the full-length M protein from Lagos bat virus (LAG M). Successful crystallization depended on a number of factors, in particular the addition of an N-terminal SUMO fusion tag to increase protein solubility. Diffraction data have been recorded from crystals of native and selenomethionine-labelled LAG M to 2.75 and 3.0 A resolution, respectively. Preliminary analysis indicates that these crystals belong to space group P6(1)22 or P6(5)22, with unit-cell parameters a = b = 56.9-57.2, c = 187.9-188.6 A, consistent with the presence of one molecule per asymmetric unit, and structure determination is currently in progress.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Chiroptera
  • Cloning, Molecular
  • Crystallization
  • Crystallography, X-Ray
  • Dogs
  • Gene Expression Regulation, Viral / physiology*
  • Lyssavirus / chemistry*
  • Lyssavirus / genetics*
  • Lyssavirus / isolation & purification
  • Molecular Sequence Data
  • Viral Matrix Proteins / biosynthesis*
  • Viral Matrix Proteins / chemistry
  • Viral Matrix Proteins / genetics*
  • Viral Matrix Proteins / isolation & purification

Substances

  • Viral Matrix Proteins