Mapping of neutralizing epitopes to fragments of the bovine coronavirus E2 protein by proteolysis of antigen-antibody complexes.
Neutralizing antigenic domains on bovine coronavirus gp100/E2 were mapped to fragments of this protein by proteolytic cleavage and fragment analysis. The procedure involved analysis of fragments generated after incubation of E2-monoclonal antibody complexes with various proteases. The smallest antibody-bound fragments obtained were a 50K fragment following Staphylococcus aureus V8 protease and submaxillary protease digestion, and a 37K fragment following trypsin digestion. Trypsin also produced a transient antibody-bound 50K fragment. A 40K fragment which was not bound by antibody was observed following digestions with all three proteases. The 50K fragments generated by V8, submaxillary protease and trypsin comigrated on gels and displayed the same altered mobility under non-reducing conditions, suggesting identity of these fragments and indicating the presence of disulphide linkages in these fragments. The 40K fragments generated by these three enzymes also comigrated and displayed the same altered mobility under non-reducing conditions. The 37K trypsin fragment contained both neutralizing domains, A and B.
Deregt
D
D
Department of Veterinary Microbiology, University of Saskatchewan, Saskatoon, Canada.
Parker
M D
MD
Cox
G C
GC
Babiuk
L A
LA
D000818:Animals; D000936:Antigen-Antibody Complex; D000956:Antigens, Viral / Q000235:genetics* / Q000276:immunology; D002417:Cattle; D003332:Coronaviridae / Q000235:genetics* / Q000276:immunology; D000939:Epitopes / Q000235:genetics* / Q000276:immunology; D009500:Neutralization Tests; D010449:Peptide Mapping; D011233:Precipitin Tests; D011863:Radioimmunoassay; D015183:Restriction Mapping; D012697:Serine Endopeptidases; D014357:Trypsin; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000276:immunology
D000936:Antigen-Antibody Complex; D000956:Antigens, Viral; D000939:Epitopes; D014759:Viral Envelope Proteins; D012697:Serine Endopeptidases; C028344:glutamyl endopeptidase; D014357:Trypsin