1887

Abstract

The human pathogen can cause atypical pneumonia through adherence to epithelial cells in the respiratory tract. The major immunogenic protein, P1, participates in the attachment of the bacteria to the host cells. To investigate the adhesion properties of P1, a recombinant protein (rP1-II) covering amino acids 1107–1518 of the P1 protein was produced. This protein inhibited the adhesion of to human HEp-2 cells, as visualized in a competitive-binding assay using immunofluorescence microscopy. Previous studies have shown that mAbs that recognize two epitopes in this region of P1 also reduce adhesion. Therefore, peptides covering these epitopes, of 8 and 13 aa, respectively, were synthesized to further investigate the adhesion region. None of these synthetic peptides reduced the binding of to the receptors on the host cells. Instead, 10 overlapping synthetic peptides covering the whole of rP1-II were evaluated in the competitive-binding assay using immunofluorescence microscopy. A reduction in the number of microcolonies was seen, which was confirmed for five peptides using a POLARstar OPTIMA reader to measure fluorescence intensity. The number of microcolonies adhering to the host cells was significantly reduced by these five peptides. Further investigations showed that inhibiting peptide 7 (amino acids 1347–1396) of the major adhesin protein P1 bound directly to host receptors, suggesting that the observed -inhibiting peptides occupied HEp-2 receptors, which are otherwise available for P1-mediated adhesion.

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2007-11-01
2019-11-15
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References

  1. Baseman, J. B., Cole, R. M., Krause, D. C. & Leith, D. K. ( 1982; ). Molecular basis for cytadsorption of Mycoplasma pneumoniae. J Bacteriol 151, 1514–1522.
    [Google Scholar]
  2. Baseman, J. B., Morrison-Plummer, J., Drouillard, D., Puleo-Scheppke, B., Tryon, V. V. & Holt, S. C. ( 1987; ). Identification of a 32-kilodalton protein of Mycoplasma pneumoniae associated with hemadsorption. Isr J Med Sci 23, 474–479.
    [Google Scholar]
  3. Bredt, W. ( 1979; ). Motility. In The Mycoplasmas, pp. 141–155. Edited by M. F. Barile & S. Razin. New York: Academic Press.
  4. Chaudhry, R., Nisar, N., Hora, B., Chirasani, S. R. & Malhotra, P. ( 2005; ). Expression and immunological characterization of the carboxy-terminal region of the P1 adhesin protein of Mycoplasma pneumoniae. J Clin Microbiol 43, 321–325.[CrossRef]
    [Google Scholar]
  5. Clausen, H. F., Fedder, J., Drasbek, M., Nielsen, P. K., Toft, B., Ingerslev, H. J., Birkelund, S. & Christiansen, G. ( 2001; ). Serological investigation of Mycoplasma genitalium in infertile women. Hum Reprod 16, 1866–1874.[CrossRef]
    [Google Scholar]
  6. Dallo, S. F., Su, C. J., Horton, J. R. & Baseman, J. B. ( 1988; ). Identification of P1 gene domain containing epitope(s) mediating Mycoplasma pneumoniae cytoadherence. J Exp Med 167, 718–723.[CrossRef]
    [Google Scholar]
  7. Dallo, S. F., Kannan, T. R., Blaylock, M. W. & Baseman, J. B. ( 2002; ). Elongation factor Tu and E1 β subunit of pyruvate dehydrogenase complex act as fibronectin binding proteins in Mycoplasma pneumoniae. Mol Microbiol 46, 1041–1051.[CrossRef]
    [Google Scholar]
  8. Drasbek, M., Nielsen, P. K., Persson, K., Birkelund, S. & Christiansen, G. ( 2004; ). Immune response to Mycoplasma pneumoniae P1 and P116 in patients with atypical pneumonia analyzed by ELISA. BMC Microbiol 4, 7 [CrossRef]
    [Google Scholar]
  9. Eisenberg, D., Schwarz, E., Komaromy, M. & Wall, R. ( 1984; ). Analysis of membrane and surface protein sequences with the hydrophobic moment plot. J Mol Biol 179, 125–142.[CrossRef]
    [Google Scholar]
  10. Gabridge, M. G. & Taylor-Robinson, D. ( 1979; ). Interaction of Mycoplasma pneumoniae with human lung fibroblasts: role of receptor sites. Infect Immun 25, 455–459.
    [Google Scholar]
  11. Gerstenecker, B. & Jacobs, E. ( 1990; ). Topological mapping of the P1-adhesin of Mycoplasma pneumoniae with adherence-inhibiting monoclonal antibodies. J Gen Microbiol 136, 471–476.[CrossRef]
    [Google Scholar]
  12. Hu, P. C., Huang, C. H., Collier, A. M. & Clyde, W. A., Jr ( 1983; ). Demonstration of antibodies to Mycoplasma pneumoniae attachment protein in human sera and respiratory secretions. Infect Immun 41, 437–439.
    [Google Scholar]
  13. Jacobs, E., Schopperle, K. & Bredt, W. ( 1985; ). Adherence inhibition assay: a specific serological test for detection of antibodies to Mycoplasma pneumoniae. Eur J Clin Microbiol 4, 113–118.[CrossRef]
    [Google Scholar]
  14. Jacobs, E., Gerstenecker, B., Mader, B., Huang, C. H., Hu, P. C., Halter, R. & Bredt, W. ( 1989; ). Binding sites of attachment-inhibiting monoclonal antibodies and antibodies from patients on peptide fragments of the Mycoplasma pneumoniae adhesin. Infect Immun 57, 685–688.
    [Google Scholar]
  15. Jacobs, E., Pilatschek, A., Gerstenecker, B., Oberle, K. & Bredt, W. ( 1990; ). Immunodominant epitopes of the adhesin of Mycoplasma pneumoniae. J Clin Microbiol 28, 1194–1197.
    [Google Scholar]
  16. Jacobs, E., Bartl, A., Oberle, K. & Schiltz, E. ( 1995; ). Molecular mimicry by Mycoplasma pneumoniae to evade the induction of adherence inhibiting antibodies. J Med Microbiol 43, 422–429.[CrossRef]
    [Google Scholar]
  17. Krause, D. C. ( 1996; ). Mycoplasma pneumoniae cytadherence: unravelling the tie that binds. Mol Microbiol 20, 247–253.[CrossRef]
    [Google Scholar]
  18. Krause, D. C. ( 1998; ). Mycoplasma pneumoniae cytadherence: organization and assembly of the attachment organelle. Trends Microbiol 6, 15–18.[CrossRef]
    [Google Scholar]
  19. Krause, D. C., Leith, D. K., Wilson, R. M. & Baseman, J. B. ( 1982; ). Identification of Mycoplasma pneumoniae proteins associated with hemadsorption and virulence. Infect Immun 35, 809–817.
    [Google Scholar]
  20. Layh-Schmitt, G. & Harkenthal, M. ( 1999; ). The 40- and 90-kDa membrane proteins (ORF6 gene product) of Mycoplasma pneumoniae are responsible for the tip structure formation and P1 (adhesin) association with the Triton shell. FEMS Microbiol Lett 174, 143–149.[CrossRef]
    [Google Scholar]
  21. Leigh, S. A. & Wise, K. S. ( 2002; ). Identification and functional mapping of the Mycoplasma fermentans P29 adhesin. Infect Immun 70, 4925–4935.[CrossRef]
    [Google Scholar]
  22. Novotny, L. A., Jurcisek, J. A., Pichichero, M. E. & Bakaletz, L. O. ( 2000; ). Epitope mapping of the outer membrane protein P5-homologous fimbrin adhesin of nontypeable Haemophilus influenzae. Infect Immun 68, 2119–2128.[CrossRef]
    [Google Scholar]
  23. Razin, S. & Jacobs, E. ( 1992; ). Mycoplasma adhesion. J Gen Microbiol 138, 407–422.[CrossRef]
    [Google Scholar]
  24. Razin, S., Kahane, I., Banai, M. & Bredt, W. ( 1981; ). Adhesion of mycoplasmas to eukaryotic cells. Ciba Found Symp 80, 98–118.
    [Google Scholar]
  25. Seto, S., Kenri, T., Tomiyama, T. & Miyata, M. ( 2005; ). Involvement of P1 adhesin in gliding motility of Mycoplasma pneumoniae as revealed by the inhibitory effects of antibody under optimized gliding conditions. J Bacteriol 187, 1875–1877.[CrossRef]
    [Google Scholar]
  26. Svenstrup, H. F., Nielsen, P. K., Drasbek, M., Birkelund, S. & Christiansen, G. ( 2002; ). Adhesion and inhibition assay of Mycoplasma genitalium and M. pneumoniae by immunofluorescence microscopy. J Med Microbiol 51, 361–373.
    [Google Scholar]
  27. Thomas, R. & Brooks, T. ( 2004; ). Common oligosaccharide moieties inhibit the adherence of typical and atypical respiratory pathogens. J Med Microbiol 53, 833–840.[CrossRef]
    [Google Scholar]
  28. Tully, J. G., Rose, L. D., Whitcomb, R. F. & Wenzel, R. P. ( 1979; ). Enhanced isolation of Mycoplasma pneumoniae from throat washings with a newly modified culture medium. J Infect Dis 139, 478–482.[CrossRef]
    [Google Scholar]
  29. Waites, K. B. & Talkington, D. F. ( 2004; ). Mycoplasma pneumoniae and its role as a human pathogen. Clin Microbiol Rev 17, 697–728.[CrossRef]
    [Google Scholar]
  30. Younson, J. & Kelly, C. ( 2004; ). The rational design of an anti-caries peptide against Streptococcus mutans. Mol Divers 8, 121–126.[CrossRef]
    [Google Scholar]
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