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Analysis of murine coronavirus surface glycoprotein functions by using monoclonal antibodies
E Routledge1, R Stauber, M Pfleiderer
1Institute of Virology, Würzburg, Federal Republic of Germany.
Abstract:
The murine coronavirus surface glycoprotein gene was expressed as a fusion protein in bacteria, and the expressed protein was used to generate S protein-specific monoclonal antibodies (MAbs). Three of the MAbs, 11F, 30B, and 10G, were able to neutralize virus infectivity, and two of them, 11F and 10G, were able to block virus-induced, cell-to-cell fusion. The binding sites of the 11F, 30B, and 10G MAbs were determined by Western immunoblotting and epitope mapping. The 11F and 30B MAbs bound to sites located, respectively, between amino acids 33 to 40 and 395 to 406 in the amino-terminal (S1) subunit of the S protein, and the 10G MAb bound to a site located between amino acids 1123 and 1137 in the carboxy-terminal (S2) subunit. These data define more precisely the interactions between the S1 and S2 subunits of the murine coronavirus S protein and provide further insights into its structure and function.
Insights
Researchers developed monoclonal antibodies (MAbs) targeting the murine coronavirus S protein. These MAbs neutralize virus infectivity and block cell fusion, offering insights into coronavirus structure and function.
Area of Science:
- Virology
- Immunology
- Structural Biology
Background:
- Murine coronavirus (MCV) surface glycoprotein (S protein) is crucial for viral entry and cell-to-cell fusion.
- Understanding the S protein's structure and function is key to developing antiviral strategies.
Purpose of the Study:
- To generate and characterize monoclonal antibodies (MAbs) against the MCV S protein.
- To investigate the neutralizing and fusion-inhibiting capabilities of these MAbs.
- To map the epitope binding sites of the MAbs on the S protein.
Main Methods:
- Expression of MCV S protein as a fusion protein in bacteria.
- Generation of S protein-specific monoclonal antibodies (MAbs).
- Virus neutralization assays and cell-to-cell fusion inhibition assays.
- Western immunoblotting and epitope mapping to determine MAb binding sites.
Main Results:
- Three MAbs (11F, 30B, 10G) demonstrated virus neutralization activity.
- Two MAbs (11F, 10G) inhibited virus-induced cell-to-cell fusion.
- Epitope mapping identified specific binding sites for each MAb on the S1 and S2 subunits of the S protein.
Conclusions:
- The generated MAbs provide valuable tools for studying MCV.
- Defined MAb binding sites offer precise insights into S protein structure and subunit interactions.
- This research advances our understanding of coronavirus entry mechanisms and potential therapeutic targets.