title: Localization of VP4 neutralization sites in rotavirus by three-dimensional cryo-electron microscopy.
abstract: Three-dimensional structures of several spherical viruses have been determined by electron microscopy and X-ray crystallography. We report here the first three-dimensional structure of the complex between an intact virus and Fab fragments of a neutralizing monoclonal antibody. The antibody is against VP4, one of the two outer capsid proteins of rotaviruses. These large icosahedral viruses cause gastroenteritis in children and young animals and account for over a million human deaths annually. VP4 in these viruses has been implicated in several important functions such as cell penetration, haemagglutination, neutralization and virulence. Here we demonstrate that the surface spikes on rotavirus particles are made up of VP4. Antigenic sites are located near the distal ends of the spikes and two Fab fragments bind to each of the sixty spikes. The mass of the spike indicates that it is a dimer of VP4. The bilobed structure at the distal end of the spike may be involved in both the attachment to the cell and in viral penetration. A novel feature in the virus-Fab complex is the structural difference between the two chemically equivalent Fab fragments on each spike, which could be indicative of variations in the Fab elbow angles.
authors.affiliation: Verna and Marrs McLean Department of Biochemistry, Baylor College of Medicine, Houston, Texas 77030.
authors.forename: B V
authors.initials: BV
authors.lastname: Prasad
authors.forename: J W
authors.initials: JW
authors.lastname: Burns
authors.forename: E
authors.initials: E
authors.lastname: Marietta
authors.forename: M K
authors.initials: MK
authors.lastname: Estes
authors.forename: W
authors.initials: W
authors.lastname: Chiu
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology*; D000914:Antibodies, Viral / Q000276:immunology*; D000956:Antigens, Viral / Q000276:immunology*; D001666:Binding Sites, Antibody; D005615:Freezing; D007140:Immunoglobulin Fab Fragments / Q000276:immunology*; D008854:Microscopy, Electron; D008970:Molecular Weight; D012401:Rotavirus / Q000276:immunology / Q000648:ultrastructure*
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D007140:Immunoglobulin Fab Fragments