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Home / Equine Research Bank / Clin Diagn Lab Immunol / Importance of M-protein C terminus as substrate antigen for ...
Clinical and diagnostic laboratory immunology2002; 9(3); 698-703; doi: 10.1128/cdli.9.3.698-703.2002

Importance of M-protein C terminus as substrate antigen for serodetection of equine arteritis virus infection.

Abstract: Equine arteritis virus (EAV), an enveloped positive-stranded RNA virus, is the prototype of the arterivirus group. In a previous paper (A. Kheyar, S. Martin, G. St.-Laurent, P. J. Timoney, W. H. McCollum, and D. Archambault, Clin. Diagn. Lab. Immunol. 4:648-652, 1997), we have shown that the unglycosylated membrane (M) protein, which is composed of 162 amino acids (aa), is a major target of equine antibody to EAV. In order to determine the antigenic regions of the M protein, the cDNA encoding the M protein of EAV was inserted into the procaryotic expression vector pGEX-4T-1 to produce recombinant glutathione S-transferase-M fusion protein. Various deletion mutant clones, which covered the entire sequence of the M protein, were then generated by inverse PCR and expressed in Escherichia coli to examine, by a Western blot assay, the antigenic reactivity of the clone-derived truncated M proteins with sera from horses either experimentally or naturally infected with EAV. Deletion of the hydrophobic N-terminal 87 aa did not abolish immune reactivity of the protein with serum antibodies to EAV, thereby demonstrating the antigenicity of the C-terminal region (aa 88 to 162) of the M protein. Further truncations of the M-protein C-terminal domain defined particular linear epitope-containing amino acid sequence regions. However, only the M-protein C-terminal region was readily recognized by all EAV-specific horse antisera tested in this study. Based on these findings, only the M-protein C-terminal polypeptide composed of aa 88 to 162 is necessary to identify horse serum antibodies specific to the EAV M protein. Thus, this polypeptide might be useful for serodetection of EAV-infected animals.
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Publication Date: 2002-05-03 PubMed ID: 11986280PubMed Central: PMC119998DOI: 10.1128/cdli.9.3.698-703.2002Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research article discusses the importance of the C-terminal region of the M-protein as a target antigen for detecting Equine arteritis virus (EAV) infection in horses.

Research Overview

  • The research focuses on understanding the antigenic regions of the M-protein of the Equine arteritis virus (EAV). EAV is a positive-stranded RNA virus affecting horses.
  • Previous research has identified the unglycosylated membrane (M) protein as a major target of equine antibodies against EAV.

Methodology

  • The scientists inserted the DNA sequence encoding the virus’s M protein into a prokaryotic (bacterial) expression vector or carrier molecule to produce a recombinant M protein.
  • They generated a series of deletion mutants, which are clones with parts of the M protein sequence missing.
  • These mutants were expressed in E. coli bacteria, and their antigenic reactivity was examined through a Western blot assay, a technique used to detect specific proteins in a sample of tissue homogenate or extract.

Results and Findings

  • Findings showed that removing the hydrophobic (water-repellent) N-terminal 87 amino acids did not eliminate the protein’s immune reactivity, highlighting the antigenicity of the C-terminal part (amino acids 88 to 162) of the M protein.
  • Further truncations of this region identified particular amino acid sequence regions that contain linear epitopes, the parts of an antigen molecule to which an antibody attaches itself.
  • However, only the C-terminal region of the M protein was consistently recognized by all EAV-specific horse antisera tested, indicating its importance in the immune response.

Conclusions

  • The findings suggest that only the C-terminal polypeptide (a potential antigenic part) of the M protein is needed to detect horse serum antibodies specific to the EAV M protein, illustrating its potential as an essential tool in diagnosing EAV infection.
  • The identified segment of the M protein could be useful for developing diagnostic tests to detect EAV-infected animals clinically efficiently.

Cite This Article

APA
Jeronimo C, Archambault D. (2002). Importance of M-protein C terminus as substrate antigen for serodetection of equine arteritis virus infection. Clin Diagn Lab Immunol, 9(3), 698-703. https://doi.org/10.1128/cdli.9.3.698-703.2002

Publication

Clinical and diagnostic laboratory immunology
ISSN: 1071-412X
NlmUniqueID: 9421292
Country: United States
Language: English
Volume: 9
Issue: 3
Pages: 698-703

Researcher Affiliations

Jeronimo, Célia
  • Laboratory of Molecular Virology and Immunology, Department of Biological Sciences, University of Québec at Montréal, Succursale Centre-Ville, Montréal, Québec, Canada H3C 3P8.
Archambault, Denis

    MeSH Terms

    • Animals
    • Antibodies, Viral / blood
    • Antibodies, Viral / immunology
    • Antigens, Viral / genetics
    • Antigens, Viral / immunology
    • Arterivirus Infections / blood
    • Arterivirus Infections / immunology
    • Arterivirus Infections / veterinary
    • Arterivirus Infections / virology
    • Blotting, Western / methods
    • Epitopes, B-Lymphocyte / genetics
    • Epitopes, B-Lymphocyte / immunology
    • Equartevirus / immunology
    • Escherichia coli
    • Gene Expression
    • Glutathione Transferase / genetics
    • Horse Diseases / blood
    • Horse Diseases / diagnosis
    • Horse Diseases / immunology
    • Horse Diseases / virology
    • Horses
    • Recombinant Fusion Proteins / genetics
    • Recombinant Fusion Proteins / immunology
    • Serologic Tests
    • Viral Envelope Proteins / genetics
    • Viral Envelope Proteins / immunology

    References

    This article includes 35 references
    1. Abed Y, St.-Laurent G, Zhang H, Jacobs RM, Archambault D. Development of a Western blot assay for detection of bovine immunodeficiency-like virus using capsid and transmembrane envelope proteins expressed from recombinant baculovirus. Clin. Diagn. Lab. Immunol. 1999 6:168-172.
      pmc: PMC95681pubmed: 10066648
    2. Balasuriya UBR, MacLachlan NJ, de Vries AAF, Rossitto PV, Rottier PJM. Identification of a neutralization site in the major glycoprotein (GL) of equine arteritis virus. Virology 1995 207:518-527.
      pubmed: 7533965
    3. Balasuriya UBR, Timoney PJ, McCollum WH, MacLachlan NJ. Phylogenetic analysis of open reading frame 5 of field isolates of equine arteritis virus and identification of conserved and nonconserved regions in the GL envelope. Virology 1995 214:690-697.
      pubmed: 8553578
    4. Bentley L, Fehrsen J, Jordaan F, Huismans H, du Plessis DH. Identification of antigenic regions on VP2 African horse sickness virus serotype 3 by using phage-displayed epitope libraries. J. Gen. Virol. 2000 81:993-1000.
      pubmed: 10725425
    5. Cavanagh D. Nidovirales: a new order comprising Coronaviridae and Arteriviridae. Arch. Virol. 1997 142:629-633.
      pubmed: 9349308
    6. Chirnside ED, Wearing CM, Binns MM, Mumford JA. Comparison of M and N gene sequences distinguishes variation amongst equine arteritis virus isolates. J. Gen. Virol. 1994 75:1491-1497.
      pubmed: 8207415
    7. Chirnside ED, de Vries AAF, Mumford JA, Rottier PJM. Equine arteritis virus-neutralizing antibody in the horses is induced by a determinant on the large envelope glycoprotein GL. J. Gen. Virol. 1995 76:1989-1998.
      pubmed: 7636479
    8. Chirnside ED, Francis PM, de Vries AAF, Sinclair R, Mumford JA. Development and evaluation of an ELISA using recombinant fusion protein to detect the presence of host antibody to equine arteritis virus. J. Virol. Methods 1995 54:1-13.
      pmc: PMC7119792pubmed: 7559853
    9. Chirnside ED, Francis PM, Mumford JA. Expression cloning and antigenic analysis of the nucleocapsid protein of the equine arteritis virus. Virus Res. 1995 39:277-288.
      pmc: PMC7133929pubmed: 8837890
    10. Cho HJ, Entz SC, Deregt D, Jordan LT, Timoney PJ, McCollum WH. Detection of antibodies to equine arteritis virus by monoclonal antibody-based blocking ELISA. Can. J. Vet. Res. 2000 64:38-43.
      pmc: PMC1189579pubmed: 10680655
    11. Dea S, Gagnon CA, Mardassi H, Milane G. Antigenic variability among North American and European strains of porcine reproductive and respiratory syndrome virus as defined by monoclonal antibodies to the matrix protein. J. Clin. Microbiol. 1996 34:1488-1493.
      pmc: PMC229047pubmed: 8735103
    12. den Boon JA, Snijder EJ, Chirnside ED, de Vries AAF, Horzinek MC, Spaan WJM. Equine arteritis virus is not a togavirus but belongs to the coronaviruslike superfamily. J. Virol. 1991 65:2910-2920.
      pmc: PMC240924pubmed: 1851863
    13. Deregt D, de Vries AAF, Raamsman MJB, Elmgren LD, Rottier PJM. Monoclonal antibodies to equine arteritis virus proteins identify the GL protein as a target for virus neutralization. J. Gen. Virol. 1994 75:2439-2444.
      pubmed: 8077945
    14. de Vries AAF, Chirnside ED, Gravestein LA, Horzinek MC, Spaan WJ. All subgenomic mRNAs of equine arteritis virus contain a common leader sequence. Nucleic Acids Res. 1990 18:3241-3247.
      pmc: PMC330929pubmed: 2162519
    15. de Vries AAF, Chirnside ED, Horzinek MC, Rottier PJM. Structural proteins of equine arteritis virus. J. Virol. 1992 66:6294-6303.
      pmc: PMC240121pubmed: 1328669
    16. Doll ER, Bryans JT, McCollum WH, Crowe MEW. Isolation of a filterable agent causing arteritis of horses and abortion by mares. Its differentiation from the equine abortion (influenza) virus. Cornell Vet. 1957 47:3-41.
      pubmed: 13397177
    17. Frangioni JV, Neel BG. Solubilization and purification of enzymatically active glutathione S-transferase (pGEX) fusion proteins. Anal. Biochem. 1993 210:179-187.
      pubmed: 8489015
    18. Geysen HM, Meloen H, Barteling SJ. Use of peptide synthesis to probe viral antigens for epitopes to a resolution of a single amino acid. Proc. Natl. Acad. Sci. USA 1984 81:3998-4002.
      pmc: PMC345355pubmed: 6204335
    19. Glaser AL, de Vries AAF, Dubovi EJ. Comparison of equine arteritis virus isolates using neutralizing monoclonal antibodies and identification of sequence changes in GL associated with neutralization resistance. J. Gen. Virol. 1995 76:2223-2233.
      pubmed: 7561759
    20. Hedges JF, Balasuriya UBR, Ahmad S, Timoney PJ, McCollum WH, Yilma T, MacLachlan NJ. Detection of antibodies to equine arteritis virus by enzyme linked immunosorbant assays utilizing GL, M and N proteins expressed from recombinant baculoviruses. J. Virol. Methods 1998 76:127-137.
      pubmed: 9923747
    21. Hedges JF, Balasuriya UBR, MacLachlan NJ. The open reading frame 3 of equine arteritis virus encodes an immunogenic glycosylated integral membrane protein. Virology 1999 264:92-98.
      pubmed: 10544133
    22. Hidajat R, McNicol P. Primer-directed mutagenesis of an intact plasmid by using Pwo DNA polymerase in long distance inverse PCR. BioTechniques 1997 22:33-34.
      pubmed: 8994640
    23. Kheyar A, Martin S, St.-Laurent G, Timoney PJ, McCollum WH, Archambault D. Expression cloning and humoral immune response to the nucleocapsid and membrane proteins of equine arteritis virus. Clin. Diagn. Lab. Immunol. 1997 4:648-652.
      pmc: PMC170634pubmed: 9384283
    24. MacLachlan NJ, Balasuriya UBR, Hedges JF, Schweidler TM, McCollum WH, Timoney PJ, Hullinger PJ, Patton JF. Serologic response of horses to the structural proteins of equine arteritis virus. J. Vet. Diagn. Investig. 1998 10:229-236.
      pubmed: 9683071
    25. Maeng C-Y, Ryu CJ, Gripon P, Guguen-Guillouzo C, Hong HJ. Fine mapping of virus-neutralizing epitopes on hepatitis B virus preS1. Virology 2000 270:9-16.
      pubmed: 10772975
    26. Magar R, Larochelle R, Nelson EA, Charreyre C. Differential reactivity of a monoclonal antibody directed to the membrane protein of porcine reproductive and respiratory syndrome virus. Can. J. Vet. Res. 1997 61:69-71.
      pmc: PMC1189374pubmed: 9008806
    27. Megret F, Hugnot JP, Falconar A, Gentry MK, Morens DM, Murray JM, Schlesinger JJ, Wright PJ, Young P, Van Regenmortel MHV, Deubel V. Use of recombinant fusion proteins and monoclonal antibodies to define linear and discontinuous antigenic sites on the dengue virus envelope glycoprotein. Virology 1992 187:480-491.
      pubmed: 1372140
    28. Nugent J, Sinclair R, de Vries AA, Eberhardt RY, Castillo-Olivares J, Davis Poynter N, Rottier PJ, Mumford JA. Development and evaluation of ELISA procedures to detect antibodies against the major envelope protein (G(L)) of equine arteritis virus. J. Virol. Methods 2000 90:167-183.
      pubmed: 11064117
    29. Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc. Natl. Acad. Sci. USA 1977 74:5463-5467.
      pmc: PMC431765pubmed: 271968
    30. Senne DA, Pearson JE, Carbrey EA. Equine viral arteritis: a standard procedure for the virus neutralization test and comparison of results of a proficiency test performed at five laboratories, p. 29-34. In Proceedings of the 89th Annual Meeting of the United States Animal Health Association, Milwaukee, Wisconsin. .
    31. Snijder EJ, Meulenberg JM. The molecular biology of arteriviruses. J. Gen. Virol. 1998 79:961-979.
      pubmed: 9603311
    32. Snijder EJ, van Toll H, Pedersen W, Raamsman MJB, de Vries AAF. Identification of a novel structural protein of arteriviruses. J. Virol. 1999 73:6335-6345.
      pmc: PMC112712pubmed: 10400725
    33. Stadejek T, Björklund H, Ros Bascuñana C, Ciabatti IM, Scicluna MT, Amaddeo D, McCollum WH, Autorino GL, Timoney PJ, Paton DJ, Klingeborn B, Belák S. Genetic diversity of equine arteritis virus. J. Gen. Virol. 1999 80:691-699.
      pubmed: 10092009
    34. Timoney PJ, McCollum WH. Equine viral arteritis. Vet. Clin. N. Am. Equine Pract. 1993 9:295-309.
      pmc: PMC7134676pubmed: 8395325
    35. Zaripov MM, Morenkov OS, Fodor N, Brown A, Schmatchenco VV, Fodor I. Distribution of B-cell epitopes on the pseudorabies virus glycoprotein B. J. Gen. Virol. 1999 80:537-541.
      pubmed: 10091990

    Citations

    This article has been cited 5 times.
    1. Valle-Casuso JC, Gaudaire D, Martin-Faivre L, Madeline A, Dallemagne P, Pronost S, Munier-Lehmann H, Zientara S, Vidalain PO, Hans A. Replication of Equine arteritis virus is efficiently suppressed by purine and pyrimidine biosynthesis inhibitors. Sci Rep 2020 Jun 22;10(1):10100.
      doi: 10.1038/s41598-020-66944-4pubmed: 32572069google scholar: lookup
    2. Balasuriya UB, Go YY, MacLachlan NJ. Equine arteritis virus. Vet Microbiol 2013 Nov 29;167(1-2):93-122.
      doi: 10.1016/j.vetmic.2013.06.015pubmed: 23891306google scholar: lookup
    3. Go YY, Snijder EJ, Timoney PJ, Balasuriya UB. Characterization of equine humoral antibody response to the nonstructural proteins of equine arteritis virus. Clin Vaccine Immunol 2011 Feb;18(2):268-79.
      doi: 10.1128/CVI.00444-10pubmed: 21147938google scholar: lookup
    4. Go YY, Wong SJ, Branscum AJ, Demarest VL, Shuck KM, Vickers ML, Zhang J, McCollum WH, Timoney PJ, Balasuriya UB. Development of a fluorescent-microsphere immunoassay for detection of antibodies specific to equine arteritis virus and comparison with the virus neutralization test. Clin Vaccine Immunol 2008 Jan;15(1):76-87.
      doi: 10.1128/CVI.00388-07pubmed: 18032597google scholar: lookup
    5. Archambault D, St-Louis MC, Martin S. Binding of cellular proteins to the leader RNA of equine arteritis virus. Virus Genes 2005 Jan;30(1):121-5.
      doi: 10.1007/s11262-004-4589-6pubmed: 15744570google scholar: lookup
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