Analysis of an immunodominant region of infectious bronchitis virus.
We analyzed the antigenic fine-structure of an immunodominant region in the peplomer protein of infectious bronchitis virus. This region near the N-terminus of the S2 subunit is recognized by polyclonal antisera and by the majority of mAb that cross-react with denatured protein. Despite their involvement in neutralization, epitopes in this region were conserved in different serotypes. Epitopes of four mAb and two chicken antisera were localized by using prokaryotic expression of cDNA fragments, and overlapping peptides with lengths increasing from 3 to 12 residues (PEPSCAN). We found overlapping epitopes with lengths of 6, 9, 11, and more than 17 residues. The results indicate that the expression products are antigenically equivalent to denatured protein fragments. This suggests a general strategy for the localization of sequential epitopes in large proteins. We propose that the immunodominance of the N-terminal region of S2 is explained by features of the protein structure. Presumably, this region is a protruding protein segment of about 20 residues with a high local mobility, as indicated by the antigenicity of the peptides. The conservation of the sequence points to an involvement in a molecular recognition process during infection.
Kusters
J G
JG
Institute of Molecular Biology, University of Utrecht, The Netherlands.
Jager
E J
EJ
Lenstra
J A
JA
Koch
G
G
Posthumus
W P
WP
Meloen
R H
RH
van der Zeijst
B A
BA
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D001666:Binding Sites, Antibody; D003332:Coronaviridae / Q000276:immunology*; D000939:Epitopes / Q000235:genetics / Q000302:isolation & purification*; D001351:Infectious bronchitis virus / Q000235:genetics / Q000276:immunology*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000235:genetics / Q000276:immunology; D010449:Peptide Mapping; D010455:Peptides / Q000138:chemical synthesis; D011487:Protein Conformation; D013329:Structure-Activity Relationship; D014764:Viral Proteins / Q000235:genetics / Q000302:isolation & purification*
D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D000939:Epitopes; D010446:Peptide Fragments; D010455:Peptides; D014764:Viral Proteins