Use of substituted and tandem‐repeated peptides to probe the relevance of the highly conserved RGD tripeptide in the immune response against foot‐and‐mouth disease virus

FEBS Letters

Andreu, David²  Mateu, Mauricio G.¹  Giralt, Ernest²  Domingo, Esteban¹  Novella, Isabel S.²  Borrego, Belén¹ 

[1]Severo Ochoa Center for Molecular Biology, Cantoblanco, 28049 Madrid, Spain

[2]Department of Organic Chemistry, University of Barcelona, 08028 Barcelona, Spain

关键词: Immunopeptide;    RGD;    Cross-neutralization;    Antigenic variability;    EID;    enzyme-linked immunodot assay;    EITB;    enzyme linked immunoelectrotransfer blot assay;    FMDV;    foot-and-mouth disease virus;    TFA;    trifluoroacetic acid;    MBS;    3-maleimidobenzoyl-N-hydroxysuccinimide ester;    SPDP;    N-succinimidyl-3-(2-pyridyldithio)-propionate;   

DOI  :  10.1016/0014-5793(93)80883-V

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Antigenic site A of foot-and-mouth disease virus (FMDV) is an exposed, mobile loop which includes a central, highly conserved Arg-Gly-Asp tripeptide (RGD, VP1 residues 141–143 in serotype C) thought to be part of the cell attachment site. We have analyzed the contribution of RGD to the interaction of site A with antibodies by incorporating selected amino acid replacements at RGD into synthetic peptides representing site A, and analyzing the reactivity of substituted peptides with site A-specific monoclonal antibodies (MAbs). Replacement of Arg-141, Gly-142 or Asp-143 by alanine resulted in the loss of one, three and five epitopes, respectively, out of seven epitopes probed. Other replacements resulted in the loss of even larger numbers of epitopes, suggesting that the amino acids of the RGD region are either directly involved in interaction with antibodies or that they exert an important influence on the interaction of surrounding residues with antibodies. Thus, we explored the ability of tandem repeats of the RGDL sequence (corresponding to FMDV C-S8cl) to evoke neutralizing antibodies in rabbits and guinea pigs. Neutralizing activity was generally low but with a broad specificity for different FMDV serotypes and variants. Significant decreases in neutralizing titers were observed with boosting, suggesting a possible suppression of those anti-peptide antibodies which may also be directed to cellular RGD sequences. The results point to an involvement of RGD in the antigenic structure of site A, and open the possibility that broadly neutralizing antibodies might be induced by tandem repeats of the critical, conserved domain.

【 授权许可】

Unknown