Staff Profile
Publication details for Dr Paul Yeo
Murray, J, Loney, C, Murphy, LB, Graham, S & Yeo, RP (2001). Characterization of Monoclonal Antibodies raised against Recombinant Respiratory Syncytial Virus Nucleocapsid (N) Protein: Identification of a Region in the Carboxy Terminus of N Involved in the Interaction with P Protein. Virology 289(2): 252-261.- Publication type: Journal papers: academic
- ISSN/ISBN: 0042-6822
- DOI: 10.1006/viro.2001.1150
- Keywords: RSV; nucleocapsid; N–P interaction; monoclonal antibodies; peptides; P binding.
- View online: Online version
- Durham research online: DRO record
Author(s) from Durham
Abstract
To investigate structure and biological properties of the nucleocapsid (N) protein of respiratory syncytial virus (RSV), we have generated a panel of 16 monoclonal antibodies, raised against recombinant N protein, and epitope mapped seven of these to three antigenic sites (Site I aa 16–30; Site II aa 341–350; Site III aa 351–365). Characterization by immunofluorescence and by immunoprecipitation assay demonstrated that a monoclonal antibody to antigenic site I can detect N protein complexed with phospho (P) protein. Antibodies to antigenic sites II and III, which are adjacent to each other near the carboxyl terminus of the N protein, have distinct properties. A site III monoclonal antibody detected N protein in cytoplasmic inclusion bodies and in the cytosol, but not when N was complexed to P protein, while the site II antibody reacted with N protein in the nucleocapsid fraction but did not detect cytosolic N protein. Further investigation into the reactivities of the antibodies after binding of P to N in vitro demonstrated that antigenic sites II and III were blocked by the interaction, indicating an involvement for the carboxy domain of N in the N–P interaction. This was confirmed by the ability of peptides from the carboxy terminus of N to inhibit the N–P interaction in vitro.
