Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1989 Dec;57(12):3720–3726. doi: 10.1128/iai.57.12.3720-3726.1989

Characterization of the Pseudomonas aeruginosa pilus adhesin: confirmation that the pilin structural protein subunit contains a human epithelial cell-binding domain.

R T Irvin 1, P Doig 1, K K Lee 1, P A Sastry 1, W Paranchych 1, T Todd 1, R S Hodges 1
PMCID: PMC259896  PMID: 2572560

Abstract

Previous studies have suggested that the Pseudomonas aeruginosa PAK pilus adhesin moiety resides in an epithelial cell-binding domain located in the C-terminal region of the PAK pilin structural protein. Synthetic peptides Ac17red (a synthetic peptide with a sequence identical to that of PAK pilin residues 128 to 144, with the Cys-129 and Cys-142 residues being in the reduced state) and Ac17ox (a synthetic peptide with a sequence identical to that of PAK pilin residues 128 to 144, with a formed disulfide bridge between the amino acid residues Cys-129 and Cys-142), which should contain the epithelial cell-binding domain, were synthesized. Ac17red and Ac17ox both bound to buccal epithelial cells (BECs) and to ciliated tracheal epithelial cells (TECs). Ac17ox had a Km of 6.40 microM for binding to BECs, while Ac17red had a Km of 9.87 microM. Ac17red bound to the same receptor sites that purified pili did and competitively inhibited the binding of purified PAK pili to BECs. BEC glycoproteins with molecular masses of 82, 55 to 51, and 40 kilodaltons immobilized on nitrocellulose exhibited periodate-sensitive receptor activity for Ac17red; similar activity has been found for PAK pili. Ac17red, Ac17ox, and PAK pili bound to the cilia and luminal portions of the cytoplasmic membrane of human TECs, the same regions to which P. aeruginosa whole cells bind. PAK pilin has an epithelial cell-binding domain that resides in the C-terminal region of the protein.

Full text

PDF
3725

Images in this article

3722Image
on p.3722
3723Image
on p.3723
3724Image
on p.3724

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abraham S. N., Goguen J. D., Sun D., Klemm P., Beachey E. H. Identification of two ancillary subunits of Escherichia coli type 1 fimbriae by using antibodies against synthetic oligopeptides of fim gene products. J Bacteriol. 1987 Dec;169(12):5530–5536. doi: 10.1128/jb.169.12.5530-5536.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bradley D. E., Pitt T. L. Pilus-dependence of four Pseudomonas aeruginosa bacteriophages with non-contractile tails. J Gen Virol. 1974 Jul;24(1):1–15. doi: 10.1099/0022-1317-24-1-1. [DOI] [PubMed] [Google Scholar]
  3. Doig P., Smith N. R., Todd T., Irvin R. T. Characterization of the binding of Pseudomonas aeruginosa alginate to human epithelial cells. Infect Immun. 1987 Jun;55(6):1517–1522. doi: 10.1128/iai.55.6.1517-1522.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Franklin A. L., Todd T., Gurman G., Black D., Mankinen-Irvin P. M., Irvin R. T. Adherence of Pseudomonas aeruginosa to cilia of human tracheal epithelial cells. Infect Immun. 1987 Jun;55(6):1523–1525. doi: 10.1128/iai.55.6.1523-1525.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Garibaldi R. A., Britt M. R., Coleman M. L., Reading J. C., Pace N. L. Risk factors for postoperative pneumonia. Am J Med. 1981 Mar;70(3):677–680. doi: 10.1016/0002-9343(81)90595-7. [DOI] [PubMed] [Google Scholar]
  6. Hazlett L. D., Moon M. M., Strejc M., Berk R. S. Evidence for N-acetylmannosamine as an ocular receptor for P. aeruginosa adherence to scarified cornea. Invest Ophthalmol Vis Sci. 1987 Dec;28(12):1978–1985. [PubMed] [Google Scholar]
  7. Hazlett L. D., Moon M., Berk R. S. In vivo identification of sialic acid as the ocular receptor for Pseudomonas aeruginosa. Infect Immun. 1986 Feb;51(2):687–689. doi: 10.1128/iai.51.2.687-689.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hodges R. S., Saund A. K., Chong P. C., St-Pierre S. A., Reid R. E. Synthetic model for two-stranded alpha-helical coiled-coils. Design, synthesis, and characterization of an 86-residue analog of tropomyosin. J Biol Chem. 1981 Feb 10;256(3):1214–1224. [PubMed] [Google Scholar]
  9. Huxley E. J., Viroslav J., Gray W. R., Pierce A. K. Pharyngeal aspiration in normal adults and patients with depressed consciousness. Am J Med. 1978 Apr;64(4):564–568. doi: 10.1016/0002-9343(78)90574-0. [DOI] [PubMed] [Google Scholar]
  10. Johanson W. G., Jr, Higuchi J. H., Chaudhuri T. R., Woods D. E. Bacterial adherence to epithelial cells in bacillary colonization of the respiratory tract. Am Rev Respir Dis. 1980 Jan;121(1):55–63. doi: 10.1164/arrd.1980.121.1.55. [DOI] [PubMed] [Google Scholar]
  11. Johanson W. G., Jr, Woods D. E., Chaudhuri T. Association of respiratory tract colonization with adherence of gram-negative bacilli to epithelial cells. J Infect Dis. 1979 Jun;139(6):667–673. doi: 10.1093/infdis/139.6.667. [DOI] [PubMed] [Google Scholar]
  12. Klemm P., Christiansen G. Three fim genes required for the regulation of length and mediation of adhesion of Escherichia coli type 1 fimbriae. Mol Gen Genet. 1987 Jul;208(3):439–445. doi: 10.1007/BF00328136. [DOI] [PubMed] [Google Scholar]
  13. Laemmli U. K., Favre M. Maturation of the head of bacteriophage T4. I. DNA packaging events. J Mol Biol. 1973 Nov 15;80(4):575–599. doi: 10.1016/0022-2836(73)90198-8. [DOI] [PubMed] [Google Scholar]
  14. Lund B., Lindberg F., Marklund B. I., Normark S. The PapG protein is the alpha-D-galactopyranosyl-(1----4)-beta-D-galactopyranose-binding adhesin of uropathogenic Escherichia coli. Proc Natl Acad Sci U S A. 1987 Aug;84(16):5898–5902. doi: 10.1073/pnas.84.16.5898. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lund B., Lindberg F., Normark S. Structure and antigenic properties of the tip-located P pilus proteins of uropathogenic Escherichia coli. J Bacteriol. 1988 Apr;170(4):1887–1894. doi: 10.1128/jb.170.4.1887-1894.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Marcus H., Baker N. R. Quantitation of adherence of mucoid and nonmucoid Pseudomonas aeruginosa to hamster tracheal epithelium. Infect Immun. 1985 Mar;47(3):723–729. doi: 10.1128/iai.47.3.723-729.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Matsoukas J. M., Scanlon M. N., Moore G. J. A cyclic angiotensin antagonist: [1,8-cysteine]angiotensin II. J Med Chem. 1984 Mar;27(3):404–406. doi: 10.1021/jm00369a030. [DOI] [PubMed] [Google Scholar]
  18. McEachran D. W., Irvin R. T. Adhesion of Pseudomonas aeruginosa to human buccal epithelial cells: evidence for two classes of receptors. Can J Microbiol. 1985 Jun;31(6):563–569. doi: 10.1139/m85-105. [DOI] [PubMed] [Google Scholar]
  19. Paranchych W., Sastry P. A., Frost L. S., Carpenter M., Armstrong G. D., Watts T. H. Biochemical studies on pili isolated from Pseudomonas aeruginosa strain PAO. Can J Microbiol. 1979 Oct;25(10):1175–1181. doi: 10.1139/m79-182. [DOI] [PubMed] [Google Scholar]
  20. Paranchych W., Sastry P. A., Volpel K., Loh B. A., Speert D. P. Fimbriae (pili): molecular basis of Pseudomonas aeruginosa adherence. Clin Invest Med. 1986;9(2):113–118. [PubMed] [Google Scholar]
  21. Ramphal R., Pier G. B. Role of Pseudomonas aeruginosa mucoid exopolysaccharide in adherence to tracheal cells. Infect Immun. 1985 Jan;47(1):1–4. doi: 10.1128/iai.47.1.1-4.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Ramphal R., Sadoff J. C., Pyle M., Silipigni J. D. Role of pili in the adherence of Pseudomonas aeruginosa to injured tracheal epithelium. Infect Immun. 1984 Apr;44(1):38–40. doi: 10.1128/iai.44.1.38-40.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Rivera M., Nicotra M. B. Pseudomonas aeruginosa mucoid strain. Its significance in adult chest diseases. Am Rev Respir Dis. 1982 Nov;126(5):833–836. doi: 10.1164/arrd.1982.126.5.833. [DOI] [PubMed] [Google Scholar]
  24. Sato H., Okinaga K., Saito H. Role of pili in the pathogenesis of Pseudomonas aeruginosa burn infection. Microbiol Immunol. 1988;32(2):131–139. doi: 10.1111/j.1348-0421.1988.tb01372.x. [DOI] [PubMed] [Google Scholar]
  25. Speert D. P. Host defenses in patients with cystic fibrosis: modulation by Pseudomonas aeruginosa. Surv Synth Pathol Res. 1985;4(1):14–33. doi: 10.1159/000156962. [DOI] [PubMed] [Google Scholar]
  26. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Woods D. E., Bass J. A., Johanson W. G., Jr, Straus D. C. Role of adherence in the pathogenesis of Pseudomonas aeruginosa lung infection in cystic fibrosis patients. Infect Immun. 1980 Dec;30(3):694–699. doi: 10.1128/iai.30.3.694-699.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Woods D. E., Straus D. C., Johanson W. G., Jr, Berry V. K., Bass J. A. Role of pili in adherence of Pseudomonas aeruginosa to mammalian buccal epithelial cells. Infect Immun. 1980 Sep;29(3):1146–1151. doi: 10.1128/iai.29.3.1146-1151.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Woodward M. P., Young W. W., Jr, Bloodgood R. A. Detection of monoclonal antibodies specific for carbohydrate epitopes using periodate oxidation. J Immunol Methods. 1985 Apr 8;78(1):143–153. doi: 10.1016/0022-1759(85)90337-0. [DOI] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES