title: Rotavirus spike structure and polypeptide composition.
abstract: Negatively stained preparations of rotavirus imaged with a low dose of electrons provide sufficient contrast to reveal surface projections or spikes. The number of spikes found projecting from different particles indicates that not all 60 peripentonal sites are occupied. Treatment at pH 11.2 with 250 mM ammonium hydroxide specifically removes the spikes, yielding smooth double-shelled particles of the same diameter as that of the native virus. Protein analysis confirms that the released spikes are composed of polypeptide VP4 (or its two cleavage products VP5* and VP8*) and that the smooth particle retains the other major outer shell protein VP7. Spikeless particles can be decorated by a monoclonal antibody specific for the major immunodominant neutralizing domain of VP7, implying that removal of the spikes does not denature the VP7 that is retained on the surface of the smooth particle.
authors.affiliation: Department of Cellular and Molecular Biology, University of Auckland, New Zealand.
authors.forename: I D
authors.initials: ID
authors.lastname: Anthony
authors.forename: S
authors.initials: S
authors.lastname: Bullivant
authors.forename: S
authors.initials: S
authors.lastname: Dayal
authors.forename: A R
authors.initials: AR
authors.lastname: Bellamy
authors.forename: J A
authors.initials: JA
authors.lastname: Berriman
mesh_terms: D064753:Ammonium Hydroxide; D002213:Capsid / Q000032:analysis*; D036022:Capsid Proteins; D002460:Cell Line; D005614:Freeze Fracturing; D006387:Hemagglutination, Viral; D006863:Hydrogen-Ion Concentration; D006878:Hydroxides / Q000494:pharmacology; D008854:Microscopy, Electron; D012401:Rotavirus / Q000032:analysis / Q000187:drug effects / Q000648:ultrastructure*; D014357:Trypsin / Q000494:pharmacology; D014771:Virion / Q000032:analysis / Q000187:drug effects / Q000648:ultrastructure
chemical_list: D036022:Capsid Proteins; D006878:Hydroxides; C060339:VP4 protein, Rotavirus; D064753:Ammonium Hydroxide; D014357:Trypsin