Identification of the gB homologues of equine herpesvirus types 1 and 4 as disulphide-linked heterodimers and their characterization using monoclonal antibodies.
Abstract: Equine herpesvirus types 1 and 4 (EHV-1 and EHV-4) labelled with [14C]glucosamine were purified from infected cell culture medium and profiles of their structural proteins were obtained that enabled identification of the major glycoproteins. Nine glycosylated polypeptides were identified for each virus. Preparations of the purified viruses each contained a glycoprotein which was linked by disulphide bonds, as determined by diagonal gel electrophoresis under reducing/non-reducing conditions. High Mr forms of this glycoprotein were detected for EHV-1 when the sample was not heated. The EHV-1 protein consisted of three polypeptides of Mr 108K, 76K and 58K and the EHV-4 protein consisted of three polypeptides of Mr 112K, 74K and 61K. Western blotting and immunoprecipitation with monoclonal antibodies confirmed that the EHV-1 gB homologue migrates with an apparent Mr of 108K (140K under non-reducing conditions) but is cleaved to give glycoproteins of 76K and 58K which are held together by disulphide bonds. The EHV-4 gB homologue consists of a 112K glycoprotein which is cleaved to give glycoproteins of 74K and 61K which are also linked by disulphide bonds.
Publication Date: 1989-05-01 PubMed ID: 2543773DOI: 10.1099/0022-1317-70-5-1161Google Scholar: Lookup
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- Comparative Study
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research paper discusses the identification and characterization of certain glycoproteins in equine herpesvirus types 1 and 4 using monoclonal antibodies. It reveals that these viruses each contain a disulphide-linked glycoprotein and details their different compositions.
Methodology
- The study involved EHV-1 and EHV-4 tagged with [14C]glucosamine, followed by purification from infected cell culture medium.
- The researchers identified nine glycosylated polypeptides for each of these viruses, revealing the protein structure.
- Diagonal gel electrophoresis was used under reducing and non-reducing conditions allowing the identification of disulphide linked glycoproteins.
Findings
- The researchers discovered high molecular weight (Mr) forms of glycoprotein for EHV-1 under non-heated conditions, signifying structural changes in different environments.
- The team found that the EHV-1 protein comprised three polypeptides of Mr 108K, 76K and 58K, and the EHV-4 protein comprised three polypeptides of Mr 112K, 74K and 61K, demonstrating slight differences between the two.
Confirmation Via Antibodies and Electrophoresis
- By using Western blotting and immunoprecipitation with monoclonal antibodies, the researchers confirmed the composition of these glycoproteins.
- The EHV-1 gB homologue was found to migrate with an apparent Mr of 108K (140K under non-reducing conditions) and is cleaved to produce glycoproteins of 76K and 58K, which are connected via disulphide bonds.
- Similarly, the EHV-4 gB homologue comprises a 112K glycoprotein which is cleaved to yield glycoproteins of 74K and 61K, also bound by disulphide bonds.
The study provides valuable insights into the protein structure of equine herpesviruses that could possibly assist future studies in understanding these viruses and potentially developing effective treatments.
Cite This Article
APA
Meredith DM, Stocks JM, Whittaker GR, Halliburton IW, Snowden BW, Killington RA.
(1989).
Identification of the gB homologues of equine herpesvirus types 1 and 4 as disulphide-linked heterodimers and their characterization using monoclonal antibodies.
J Gen Virol, 70 ( Pt 5), 1161-1172.
https://doi.org/10.1099/0022-1317-70-5-1161 Publication
Researcher Affiliations
- Department of Microbiology, University of Leeds, U.K.
MeSH Terms
- Animals
- Antibodies, Monoclonal
- Blotting, Western
- Disulfides / analysis
- Electrophoresis, Polyacrylamide Gel
- Glycoproteins / analysis
- Glycoproteins / immunology
- Herpesviridae / analysis
- Herpesvirus 1, Equid / analysis
- Herpesvirus 1, Equid / isolation & purification
- Mice
- Mice, Inbred BALB C
- Precipitin Tests
- Viral Proteins / analysis
- Viral Proteins / immunology
- Viral Structural Proteins
Citations
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