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

Murray, J.; Loney, C.; Murphy, L.B.; Graham, S.; Yeo, R.P.

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.

Citation

Murray, J., Loney, C., Murphy, L., Graham, S., & Yeo, R. (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. https://doi.org/10.1006/viro.2001.1150

• Journal Article Type: Article
• Publication Date: Oct 1, 2001
• Deposit Date: May 15, 2007
• Journal: Virology
• Print ISSN: 0042-6822
• Publisher: Elsevier
• Peer Reviewed: Not Peer Reviewed
• Volume: 289
• Issue: 2
• Pages: 252-261
• DOI: https://doi.org/10.1006/viro.2001.1150
• Keywords: RSV, Nucleocapsid, N–P interaction, Monoclonal antibodies, Peptides, P binding.
• Public URL: https://durham-repository.worktribe.com/output/1591086