title: Polyclonal and monoclonal antibodies specific for the six-helix bundle of the human respiratory syncytial virus fusion glycoprotein as probes of the protein post-fusion conformation.
abstract: Human respiratory syncytial virus (hRSV) has two major surface glycoproteins (G and F) anchored in the lipid envelope. Membrane fusion promoted by hRSV_F occurs via refolding from a pre-fusion form to a highly stable post-fusion state involving large conformational changes of the F trimer. One of these changes results in assembly of two heptad repeat sequences (HRA and HRB) into a six-helix bundle (6HB) motif. To assist in distinguishing pre- and post-fusion conformations of hRSV_F, we have prepared polyclonal (α-6HB) and monoclonal (R145) rabbit antibodies specific for the 6HB. Among other applications, these antibodies were used to explore the requirements of 6HB formation by isolated protein segments or peptides and by truncated mutants of the F protein. Site-directed mutagenesis and electron microscopy located the R145 epitope in the post-fusion hRSV_F at a site distantly located from previously mapped epitopes, extending the repertoire of antibodies that can decorate the F molecule.
authors.affiliation: Unidad de Biología Viral, Centro Nacional de Microbiología, Madrid, Spain; CIBER de Enfermedades Respiratorias, Instituto de Salud Carlos III, Majadahonda, 28220 Madrid, Spain.
authors.forename: Concepción
authors.initials: C
authors.lastname: Palomo
authors.affiliation: Unidad de Biología Viral, Centro Nacional de Microbiología, Madrid, Spain; CIBER de Enfermedades Respiratorias, Instituto de Salud Carlos III, Majadahonda, 28220 Madrid, Spain.
authors.forename: Vicente
authors.initials: V
authors.lastname: Mas
authors.affiliation: Unidad de Biología Viral, Centro Nacional de Microbiología, Madrid, Spain; CIBER de Enfermedades Respiratorias, Instituto de Salud Carlos III, Majadahonda, 28220 Madrid, Spain.
authors.forename: Mónica
authors.initials: M
authors.lastname: Vázquez
authors.affiliation: Unidad de Biología Viral, Centro Nacional de Microbiología, Madrid, Spain; CIBER de Enfermedades Respiratorias, Instituto de Salud Carlos III, Majadahonda, 28220 Madrid, Spain.
authors.forename: Olga
authors.initials: O
authors.lastname: Cano
authors.affiliation: Unidad de Microscopía Electrónica y Confocal, Centro Nacional de Microbiología, Instituto de Salud Carlos III, Majadahonda, 28220 Madrid, Spain.
authors.forename: Daniel
authors.initials: D
authors.lastname: Luque
authors.affiliation: Unidad de Microscopía Electrónica y Confocal, Centro Nacional de Microbiología, Instituto de Salud Carlos III, Majadahonda, 28220 Madrid, Spain.
authors.forename: María C
authors.initials: MC
authors.lastname: Terrón
authors.affiliation: National Institute for Medical Research, MRC, Mill Hill, London NW7 1AA, UK.
authors.forename: Lesley J
authors.initials: LJ
authors.lastname: Calder
authors.affiliation: Unidad de Biología Viral, Centro Nacional de Microbiología, Madrid, Spain; CIBER de Enfermedades Respiratorias, Instituto de Salud Carlos III, Majadahonda, 28220 Madrid, Spain. Electronic address: jmelero@isciii.es.
authors.forename: José A
authors.initials: JA
authors.lastname: Melero
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000276:immunology; D000914:Antibodies, Viral / Q000032:analysis / Q000276:immunology; D006801:Humans; D017433:Protein Structure, Secondary; D011817:Rabbits; D018357:Respiratory Syncytial Virus Infections / Q000821:virology*; D018113:Respiratory Syncytial Virus, Human / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D014760:Viral Fusion Proteins / Q000737:chemistry* / Q000235:genetics / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; C526850:F protein, human respiratory syncytial virus; D014760:Viral Fusion Proteins
keywords: Post-fusion specific antibodies against RSV F glycoprotein