Fine mapping of a continuous epitope on VP7 of bluetongue virus using overlapping synthetic peptides and a random epitope library.
Two complementary techniques have been used to delineate an epitope on VP7 of bluetongue virus. Two MAbs (F10 and D11), both of which bound within a region spanning amino acids 255 to 274 in the 349 amino acid protein, were used to probe overlapping synthetic peptides covering this region. A pentapeptide, QYPAL, and a hexapeptide, QY-PALT (amino acids 259-264), preferentially bound both MAbs. MAb F10 also reacted with a heptapeptide (TAEIFNV) immediately adjacent to QYPALT. The MAbs were also used to affinity-purify fusion phages from a random hexapeptide library. All phage peptides selected were similar to QYPALT. Comparison of the peptides suggested that residues Q and P at positions 1 and 3 were critical for recognition. Some affinity-purified phages displayed the hexapeptide QYPSLL, which is similar to a sequence in VP7 of another orbivirus, epizootic hemorrhagic disease virus. This finding allowed a potentially cross-reactive site to be identified.
du Plessis
D H
DH
Biochemistry Division, Onderstepoort Veterinary Institute, Republic of South Africa.
Wang
L F
LF
Jordaan
F A
FA
Eaton
B T
BT
D000595:Amino Acid Sequence; D000956:Antigens, Viral / Q000032:analysis* / Q000276:immunology; D001820:Bluetongue virus / Q000737:chemistry / Q000276:immunology*; D002213:Capsid / Q000032:analysis / Q000276:immunology*; D036022:Capsid Proteins; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis* / Q000276:immunology; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000032:analysis / Q000235:genetics / Q000276:immunology; D010449:Peptide Mapping
D000956:Antigens, Viral; D036022:Capsid Proteins; D000939:Epitopes; D010446:Peptide Fragments; C050779:VP7 protein, Rotavirus