Open AccessExpression, purification and crystallization of a lyssavirus matrix (M) protein
Author(s) -
Assenberg René,
Delmas Olivier,
Graham Stephen C.,
Verma Anil,
Berrow Nick,
Stuart David I.,
Owens Raymond J.,
Bourhy Hervé,
Grimes Jonathan M.
Publication year - 2008
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309108004557
Subject(s) - rabies virus , viral matrix protein , lyssavirus , biology , crystallization , rhabdoviridae , transcription (linguistics) , virus , mononegavirales , matrix (chemical analysis) , microbiology and biotechnology , virology , chemistry , crystallography , paramyxoviridae , organic chemistry , chromatography , viral disease , linguistics , philosophy
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 Å resolution, respectively. Preliminary analysis indicates that these crystals belong to space group P 6 1 22 or P 6 5 22, with unit‐cell parameters a = b = 56.9–57.2, c = 187.9–188.6 Å, consistent with the presence of one molecule per asymmetric unit, and structure determination is currently in progress.