Analysis and simulation of a neutralizing epitope of transmissible gastroenteritis virus.
The amino acid sequences recognized by monoclonal antibodies (MAbs) specific for the antigenic site IV of the spike protein S of transmissible gastroenteritis virus were analyzed by PEPSCAN. All MAbs of group IV recognized peptides from the S region consisting of residues 378 to 390. In addition, the neutralizing MAbs (subgroup IV-A) also bound to peptides from the region consisting of residues 1173 to 1184 and to several other peptides with a related amino acid composition. The contribution of the individual residues of both sequences to the binding of a MAb was determined by varying the length of the peptide and by a consecutive deletion or replacement of parental residues by the 19 other amino acids. The sequence consisting of residues 326 to 558, tested as part of a cro-beta-galactosidase hybrid protein, was antigenic, but the sequence consisting of residues 1150 to 1239 was not. Furthermore, antibodies raised in rabbits against the peptide SDSSFFSYGEIPFGN (residues 377 to 391), but not those raised against the peptide VRASRQLAKDKVNEC (residues 1171 to 1185), recognized the virus and had neutralizing activity. We infer that the epitope of the neutralizing MAbs is composite and consists of the linear sequence SFFSYGEI (residues 380 to 387) with contributions of A, D, K, N, Q, or V residues from other parts of the S molecule. The complex epitope was simulated by synthesizing peptides in which the sequences consisting of residues 380 to 387 and 1176 to 1184 were combined. MAbs of subgroup IV-A recognized the combination peptides two to six times better than the individual sequences. These results may offer prospects for the development of an experimental vaccine.
Central Veterinary Institute, Lelystad, The Netherlands.
W P
WP
Posthumus
J A
JA
Lenstra
W M
WM
Schaaper
A P
AP
van Nieuwstadt
L
L
Enjuanes
R H
RH
Meloen
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000954:Antigens, Surface / Q000276:immunology; D000956:Antigens, Viral / Q000235:genetics*; D015153:Blotting, Western; D003332:Coronaviridae / Q000276:immunology*; D000939:Epitopes; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D009842:Oligopeptides / Q000138:chemical synthesis / Q000276:immunology; D011994:Recombinant Proteins / Q000276:immunology; D013329:Structure-Activity Relationship; D005760:Transmissible gastroenteritis virus / Q000276:immunology*; D014764:Viral Proteins / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D000956:Antigens, Viral; D000939:Epitopes; D009842:Oligopeptides; D011994:Recombinant Proteins; D014764:Viral Proteins; C064484:spike protein S, Transmissible gastroenteritis virus