title: Expression, purification and crystallization of a lyssavirus matrix (M) protein.
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.
authors.affiliation: Division of Structural Biology and Oxford Protein Production Facility, The Henry Wellcome Building for Genomic Medicine, Oxford University, Roosevelt Drive, Oxford OX3 7BN, England.
authors.forename: René
authors.initials: R
authors.lastname: Assenberg
authors.forename: Olivier
authors.initials: O
authors.lastname: Delmas
authors.forename: Stephen C
authors.initials: SC
authors.lastname: Graham
authors.forename: Anil
authors.initials: A
authors.lastname: Verma
authors.forename: Nick
authors.initials: N
authors.lastname: Berrow
authors.forename: David I
authors.initials: DI
authors.lastname: Stuart
authors.forename: Raymond J
authors.initials: RJ
authors.lastname: Owens
authors.forename: Hervé
authors.initials: H
authors.lastname: Bourhy
authors.forename: Jonathan M
authors.initials: JM
authors.lastname: Grimes
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002685:Chiroptera; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D004285:Dogs; D015967:Gene Expression Regulation, Viral / Q000502:physiology*; D018114:Lyssavirus / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D014763:Viral Matrix Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification
chemical_list: D014763:Viral Matrix Proteins