title: Bivalent binding of a neutralising antibody to a calicivirus involves the torsional flexibility of the antibody hinge.
abstract: The structure of a complex between rabbit haemorrhagic disease virus (RHDV) virus-like particles (VLPs) and a neutralising monoclonal antibody mAb-E3 has been determined at low resolution by cryo-electron microscopy and three-dimensional (3-D) reconstruction techniques. The atomic co-ordinates of an Fab were fitted to the cryo-electron microscope density map to produce a binding model. The VLP has a T = 3 icosahedral lattice consisting of a hollow spherical shell with 90 protruding arches. Each dimeric arch presents two mAb binding sites; however, steric hindrance between the variable domains of the Fabs prevents the occupation of both sites simultaneously. Thus the maximum mAb occupation is 50%. Once a mAb is bound to one site it may bind to either of two neighbouring sites related by a local 3-fold axis. The mAbs are bound bivalently on epitopes not related by a 2-fold symmetry axis. This binding geometry implies a torsional flexibility of the mAb hinge region, involving a 60 degrees rotation of one Fab arm with respect to the other. Owing to extreme flexibility of the hinge region, the Fc domains occupy random orientations and are not visible in the reconstruction. The bivalent attachment of mAb-E3 to RHDV suggests that the neutralisation mechanism(s) involves inhibition of viral decapsidation and/or the inhibition of binding to the receptor.
authors.affiliation: Institut de Biologie Structurale, Jean-Pierre Ebel, Grenoble, France.
authors.forename: E
authors.initials: E
authors.lastname: Thouvenin
authors.forename: S
authors.initials: S
authors.lastname: Laurent
authors.forename: M F
authors.initials: MF
authors.lastname: Madelaine
authors.forename: D
authors.initials: D
authors.lastname: Rasschaert
authors.forename: J F
authors.initials: JF
authors.lastname: Vautherot
authors.forename: E A
authors.initials: EA
authors.lastname: Hewat
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000737:chemistry* / Q000378:metabolism* / Q000648:ultrastructure; D001666:Binding Sites, Antibody; D018360:Crystallography, X-Ray; D017929:Hemorrhagic Disease Virus, Rabbit / Q000276:immunology* / Q000648:ultrastructure; D007091:Image Processing, Computer-Assisted; D051379:Mice; D008807:Mice, Inbred BALB C; D008854:Microscopy, Electron; D008958:Models, Molecular; D009500:Neutralization Tests; D011487:Protein Conformation; D011817:Rabbits; D014764:Viral Proteins / Q000737:chemistry / Q000276:immunology / Q000648:ultrastructure; D014771:Virion / Q000737:chemistry / Q000276:immunology / Q000648:ultrastructure
chemical_list: D000914:Antibodies, Viral; D014764:Viral Proteins