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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

Human respiratory syncytial virus (hRSV) has two major surface glycoproteins (G and F) anchored in the lipid envelope. Membrane fusion promoted by hRSV{sub 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{sub 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{sub F} at a site distantly located from previously mapped epitopes, extending the repertoire of antibodies that can decorate the F molecule. - Highlights: • Antibodies specific for post-fusion respiratory syncytial virus fusion protein are described. • Polyclonal antibodies were obtained in rabbit inoculated with chimeric heptad repeats. • Antibody binding required assembly of a six-helix bundle in the post-fusion protein. • A monoclonal antibody with similar structural requirementsmore » is also described. • Binding of this antibody to the post-fusion protein was visualized by electron microscopy.« less
Authors:
; ; ;  [1] ;  [2] ; ;  [3] ;  [4] ;  [1] ;  [2]
  1. Unidad de Biología Viral, Centro Nacional de Microbiología, Madrid (Spain)
  2. (Spain)
  3. Unidad de Microscopía Electrónica y Confocal, Centro Nacional de Microbiología, Instituto de Salud Carlos III, Majadahonda, 28220 Madrid (Spain)
  4. National Institute for Medical Research, MRC, Mill Hill, London NW7 1AA (United Kingdom)
Publication Date:
OSTI Identifier:
22435044
Resource Type:
Journal Article
Resource Relation:
Journal Name: Virology; Journal Volume: 460-461; Other Information: Copyright (c) 2014 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; ANCHORS; CONFORMATIONAL CHANGES; ELECTRON MICROSCOPY; GLYCOPROTEINS; LIPIDS; MEMBRANES; MOLECULES; MONOCLONAL ANTIBODIES; MUTAGENESIS; PEPTIDES; RABBITS; VIRUSES