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Microbiology Society logo Microbiology

Volume 140, Issue 4

Reactivity of antibodies to heteroclitic peptides based on the major outer-membrane protein Free

Abstract

One problem of peptide vaccines is that antibodies generated against them react poorly with the target sequence on the native protein. Using monoclonal antibodies (mAbs) to the serovar L1 type-specific epitope on the major outer-membrane protein of as our model in conjunction with the Pin Technology Epitope Scanning technique, we had previously identified the critical binding site at this epitope as DAVP. Amino acid substitution showed that AV were essential residues for binding. A series of structurally related (heteroclitic) peptides retaining AV were synthesized. Some of these were found to be much more reactive with the model mAb than peptides of cognate sequence. It was hypothesized that the DAVP peptide only approximated to the conformation of the homologous sequence in the native protein, whereas some of the flexible heteroclitic peptides produced conformations which more closely resembled the native constrained sequence. The key question was whether the most reactive heteroclitic peptide would also generate antibody capable of more efficient binding to the native protein. We therefore immunized mice with one of six heteroclitic peptides or one of two native sequence control peptides. The reactivity of these antisera with the peptide immunogens and with native chlamydial elementary bodies was then evaluated by enzyme immunoassay. Pooled antisera to two of the heteroclitic peptides reacted with significantly greater absorbance ( < 0·05) and at higher dilution with whole chlamydiae than did pooled antisera to the control peptides. This suggests that heteroclitic peptides may in some circumstances be useful to increase the reactivity of site-specific antibodies with epitopes on the native protein important for vaccine development or for serodiagnosis.

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Keyword(s): Chlamydia trachomatis , epitope mapping , major outer membrane protein , peptides and vaccines
© Society for General Microbiology 1994
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1994-04-01
2025-01-20
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References

  1. van Amerongen A., Beckers P.J.A., Plasman H.H., Schaaper W.M.M., Sauerwein R.W., Meuwissen J.H.E.T., Meloen R.H. Peptides reactive with a transmission-blocking monoclonal antibody against Plasmopodium falciparium Pfs25: 2000-fold affinity increase by PEPSCAN-based amino acid substitutions. Peptide Res 1992; 5:269–274
     [Google Scholar]
  2. Baehr W., Zhang Y.X., Joseph T., Su H., Nano F.E., Everett K.D., Caldwell H.D. Mapping antigenic domains expressed by Chlamydia trachomatis major outer membrane protein genes. Proc Natl Acad Sci USA 1988; 85:4000–4004
     [Google Scholar]
  3. Bierzynski A., Kim P.S., Baldwin R.L. A salt bridge stabilizes the helix formed by isolated C-peptide of RNase A. Proc Natl Acad Sci USA 1982; 79:2470–2474
     [Google Scholar]
  4. Caldwell H.D., Kromhout J., Schachter J. Purification and partial characterization of the major outer membrane protein of Chlamydia trachomatis. Infect lmmun 1981; 31:1161–1176
     [Google Scholar]
  5. Cheetham J.C., Raleigh D.P., Griest R.E., Redfield C., Dobson C.M., Rees A.R. Antigen mobility in the combining site of an antipeptide antibody. Proc Natl Acad Sci USA 1991; 88:7968–7972
     [Google Scholar]
  6. Conlan J.W., Clarke I.N., Ward M.E. Epitope mapping with solid-phase peptides: identification of type, subspecies, species-and genus-reactive antibody binding domains on the major outer membrane protein of Chlamydia trachomatis. Mol Microbiol 1988; 2:673–679
     [Google Scholar]
  7. van Dam G.J., Verheul A.F.M., Zigterman G.J.W., J., de Reuver M.J., Snippe H. Estimation of the avidity of antibodies in polyclonal antisera against Streptococcus pneumoniae type 3 by inhibition ELISA. Mol Immunol 1989; 26:269–274
     [Google Scholar]
  8. Dyson H.J., Ranee M., Houghten R.A., Wright P.E., Lerner R.A. The order-disorder paradox in antigen-antibody union: antipeptide antibodies as a probe for structured regions of small peptides. In Biological Organisation: Macromolecular Interactions at Nigh Resolution 1987 Edited by Burnett R.M., Vogel H.J. New York: Academic Press; pp 227–234
     [Google Scholar]
  9. Dyson H.J., Ranee M., Houghten R.A., Lerner R.A., Wright P.E. Folding of immunogenic peptide fragments of proteins in water solution. I. Sequence requirements for the formation of a reverse turn. J Mol Biol 1988; 201:161–200
     [Google Scholar]
  10. Frdman G., Yxfeldt G., Wilén-Winter B., Grttnvik K.-O., Mdrdh P.-A. Large scale growth of Chlamydia trachomatis in macrophage suspension. In 2 Proceedings of the European Society for Chlamydia Research 1992 Edited by Mardh P.-A., La Placa M., Ward M. Stockholm: Uppsala University Centre for STD Research; p 24
     [Google Scholar]
  11. Getzoff E.D., Geysen H.M., Rodda S.J., Alexander H., Tainer J.A., Lerner R.A. Mechanisms of antibody binding to a protein. Science 1987; 235:1191–1196
     [Google Scholar]
  12. Geysen H.M., Rodda S.J., Mason T.J., Tribbick G., Schoofs P.G. Strategies for epitope analysis using peptide synthesis. J Immunol Methods 1987; 102:259–274
     [Google Scholar]
  13. Jones H.M., Schachter J., Stephens R.S. Evaluation of the humoral immune response in trachoma to Chlamydia trachomatis major outer membrane proteins by sequence-defined immunoassay. J Infect Dis 1992; 166:915–919
     [Google Scholar]
  14. Pickett M.A., Ward M.E., Clarke I.N. Complete nucleotide sequence of the major outer membrane protein gene from Chlamydia trachomatis serovar LI. FEMS Microbiol Lett 1987; 42:185–190
     [Google Scholar]
  15. Rini J.M., Schulze-Gahmen U., Wilson I.A. Structural evidence for induced fit as a mechanism for antibody-antigen recognition. Science 1992; 255:959–965
     [Google Scholar]
  16. Satterthwait A.C., Arrhenius T., Hagopian F.A., Zavala F., Nussenzweig V., Lerner R.A. Conformational restriction of peptidyl immunogens with covalent replacements for the hydrogen bond. Vaccine 1988; 6:99–103
     [Google Scholar]
  17. Schachter J., Dawson C. Chlamydial Infections 1978 Littleton, Massachusetts: PSA Publishing.;
     [Google Scholar]
  18. Ward M.E. Chlamydial vaccines-future trends. J. Infect 1992; 25:11–26
     [Google Scholar]
  19. Yuan Y., Zhang Y.X., Watkins N.G., Caldwell H.D. Nucleotide and deduced amino acid sequences for the four variable domains of the major outer membrane proteins of the 15 Chlamydia trachomatis serovars. Infect lmmun 1989; 57:1040–1049
     [Google Scholar]
  20. Zhang Y.X., Stewart S., Joseph T., Taylor H.R., Caldwell H.D. Protective monoclonal antibodies recognize epitopes located on the major outer membrane protein of Chlamydia trachomatis. J Immunol 1987; 138:575–581
     [Google Scholar]
/content/journal/micro/10.1099/00221287-140-4-815
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Reactivity of antibodies to heteroclitic peptides based on the Chlamydia trachomatis major outer-membrane protein
Microbiology 140, 815 (1994); https://doi.org/10.1099/00221287-140-4-815
/content/journal/micro/10.1099/00221287-140-4-815
/content/journal/micro/10.1099/00221287-140-4-815
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