Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1987 Dec;169(12):5530–5536. doi: 10.1128/jb.169.12.5530-5536.1987

Identification of two ancillary subunits of Escherichia coli type 1 fimbriae by using antibodies against synthetic oligopeptides of fim gene products.

S N Abraham 1, J D Goguen 1, D Sun 1, P Klemm 1, E H Beachey 1
PMCID: PMC213982  PMID: 2890622

Abstract

We have chemically synthesized oligopeptides corresponding to the NH2-terminal stretch of two gene products, designated FimG and FimH, of the fim gene cluster of Escherichia coli. These synthetic peptides, designated S-T1FimG(1-16) and S-T1FimH(1-25)C, evoked antibodies in rabbits that reacted with 14- and 29-kilodalton subunits, respectively, of dissociated fimbriae encoded by the recombinant plasmid pSH2 carrying the genetic information for the synthesis and expression of functional type 1 fimbriae. Neither of these fimbrial proteins was detected in dissociated fimbrial preparations from nonadhesive E. coli cells carrying the mutant plasmid pUT2002, containing a restriction site-specific deletion of fimG and fimH. Anti-S-T1FimH(1-25)C inhibited the adherence of type 1 fimbriated E. coli to epithelial cells. Immunoelectron microscopy revealed that anti-S-T1FimH(1-25)C, but not anti-S-T1FimG(1-16), bound to intact type 1 fimbriae of E. coli at the fimbrial tips and at long intervals along the fimbrial filaments. Anti-S-T1FimG(1-16) appeared to be directed at epitopes not accessible on the intact fimbriae and consequently failed to bind to intact fimbriae or to block fimbrial attachment. Our results suggest that the fimG and fimH gene products are components of type 1 fimbriae and that FimH may be the tip adhesin mediating the binding of type 1 fimbriated E. coli to D-mannose residues on mucosal surfaces.

Full text

PDF
5536

Images in this article

5532Image
on p.5532
5532Image
on p.5532
5534Image
on p.5534

Selected References

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

  1. Abraham S. N., Babu J. P., Giampapa C. S., Hasty D. L., Simpson W. A., Beachey E. H. Protection against Escherichia coli-induced urinary tract infections with hybridoma antibodies directed against type 1 fimbriae or complementary D-mannose receptors. Infect Immun. 1985 Jun;48(3):625–628. doi: 10.1128/iai.48.3.625-628.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Abraham S. N., Beachey E. H. Assembly of a chemically synthesized peptide of Escherichia coli type 1 fimbriae into fimbria-like antigenic structures. J Bacteriol. 1987 Jun;169(6):2460–2465. doi: 10.1128/jb.169.6.2460-2465.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Abraham S. N., Hasty D. L., Simpson W. A., Beachey E. H. Antiadhesive properties of a quaternary structure-specific hybridoma antibody against type 1 fimbriae of Escherichia coli. J Exp Med. 1983 Oct 1;158(4):1114–1128. doi: 10.1084/jem.158.4.1114. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Beachey E. H., Seyer J. M. Protective and nonprotective epitopes of chemically synthesized peptides of the NH2-terminal region of type 6 streptococcal M protein. J Immunol. 1986 Mar 15;136(6):2287–2292. [PubMed] [Google Scholar]
  5. Brinton C. C., Jr The structure, function, synthesis and genetic control of bacterial pili and a molecular model for DNA and RNA transport in gram negative bacteria. Trans N Y Acad Sci. 1965 Jun;27(8):1003–1054. doi: 10.1111/j.2164-0947.1965.tb02342.x. [DOI] [PubMed] [Google Scholar]
  6. Chou P. Y., Fasman G. D. Prediction of the secondary structure of proteins from their amino acid sequence. Adv Enzymol Relat Areas Mol Biol. 1978;47:45–148. doi: 10.1002/9780470122921.ch2. [DOI] [PubMed] [Google Scholar]
  7. Dale J. B., Beachey E. H. Localization of protective epitopes of the amino terminus of type 5 streptococcal M protein. J Exp Med. 1986 May 1;163(5):1191–1202. doi: 10.1084/jem.163.5.1191. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dale J. B., Seyer J. M., Beachey E. H. Type-specific immunogenicity of a chemically synthesized peptide fragment of type 5 streptococcal M protein. J Exp Med. 1983 Nov 1;158(5):1727–1732. doi: 10.1084/jem.158.5.1727. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dodd D. C., Eisenstein B. I. Antigenic quantitation of type 1 fimbriae on the surface of Escherichia coli cells by an enzyme-linked immunosorbent inhibition assay. Infect Immun. 1982 Nov;38(2):764–773. doi: 10.1128/iai.38.2.764-773.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Edman P., Begg G. A protein sequenator. Eur J Biochem. 1967 Mar;1(1):80–91. doi: 10.1007/978-3-662-25813-2_14. [DOI] [PubMed] [Google Scholar]
  11. Eshdat Y., Silverblatt F. J., Sharon N. Dissociation and reassembly of Escherichia coli type 1 pili. J Bacteriol. 1981 Oct;148(1):308–314. doi: 10.1128/jb.148.1.308-314.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Freitag C. S., Eisenstein B. I. Genetic mapping and transcriptional orientation of the fimD gene. J Bacteriol. 1983 Dec;156(3):1052–1058. doi: 10.1128/jb.156.3.1052-1058.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hopp T. P., Woods K. R. Prediction of protein antigenic determinants from amino acid sequences. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3824–3828. doi: 10.1073/pnas.78.6.3824. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hull R. A., Gill R. E., Hsu P., Minshew B. H., Falkow S. Construction and expression of recombinant plasmids encoding type 1 or D-mannose-resistant pili from a urinary tract infection Escherichia coli isolate. Infect Immun. 1981 Sep;33(3):933–938. doi: 10.1128/iai.33.3.933-938.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Klemm P., Jørgensen B. J., van Die I., de Ree H., Bergmans H. The fim genes responsible for synthesis of type 1 fimbriae in Escherichia coli, cloning and genetic organization. Mol Gen Genet. 1985;199(3):410–414. doi: 10.1007/BF00330751. [DOI] [PubMed] [Google Scholar]
  17. Klemm P. The fimA gene encoding the type-1 fimbrial subunit of Escherichia coli. Nucleotide sequence and primary structure of the protein. Eur J Biochem. 1984 Sep 3;143(2):395–399. doi: 10.1111/j.1432-1033.1984.tb08386.x. [DOI] [PubMed] [Google Scholar]
  18. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  19. Lindberg F., Lund B., Normark S. Gene products specifying adhesion of uropathogenic Escherichia coli are minor components of pili. Proc Natl Acad Sci U S A. 1986 Mar;83(6):1891–1895. doi: 10.1073/pnas.83.6.1891. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Maurer L., Orndorff P. E. Identification and characterization of genes determining receptor binding and pilus length of Escherichia coli type 1 pili. J Bacteriol. 1987 Feb;169(2):640–645. doi: 10.1128/jb.169.2.640-645.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Minion F. C., Abraham S. N., Beachey E. H., Goguen J. D. The genetic determinant of adhesive function in type 1 fimbriae of Escherichia coli is distinct from the gene encoding the fimbrial subunit. J Bacteriol. 1986 Mar;165(3):1033–1036. doi: 10.1128/jb.165.3.1033-1036.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Norgren M., Normark S., Lark D., O'Hanley P., Schoolnik G., Falkow S., Svanborg-Edén C., Båga M., Uhlin B. E. Mutations in E coli cistrons affecting adhesion to human cells do not abolish Pap pili fiber formation. EMBO J. 1984 May;3(5):1159–1165. doi: 10.1002/j.1460-2075.1984.tb01945.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Ofek I., Courtney H. S., Schifferli D. M., Beachey E. H. Enzyme-linked immunosorbent assay for adherence of bacteria to animal cells. J Clin Microbiol. 1986 Oct;24(4):512–516. doi: 10.1128/jcm.24.4.512-516.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Orndorff P. E., Falkow S. Identification and characterization of a gene product that regulates type 1 piliation in Escherichia coli. J Bacteriol. 1984 Oct;160(1):61–66. doi: 10.1128/jb.160.1.61-66.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Rothbard J. B., Fernandez R., Schoolnik G. K. Strain-specific and common epitopes of gonococcal pili. J Exp Med. 1984 Jul 1;160(1):208–221. doi: 10.1084/jem.160.1.208. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Salit I. E., Gotschlich E. C. Type I Escherichia coli pili: characterization of binding to monkey kidney cells. J Exp Med. 1977 Nov 1;146(5):1182–1194. doi: 10.1084/jem.146.5.1182. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Schifferli D. M., Abraham S. N., Beachey E. H. Influence of trimethoprim and sulfamethoxazole on the synthesis, expression, and function of type 1 fimbriae of Escherichia coli. J Infect Dis. 1986 Sep;154(3):490–496. doi: 10.1093/infdis/154.3.490. [DOI] [PubMed] [Google Scholar]
  28. Sweeney G., Freer J. H. Location of binding sites on common type 1 fimbriae from Escherichia coli. J Gen Microbiol. 1979 Jun;112(2):321–328. doi: 10.1099/00221287-112-2-321. [DOI] [PubMed] [Google Scholar]
  29. 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]
  30. Watts T. H., Sastry P. A., Hodges R. S., Paranchych W. Mapping of the antigenic determinants of Pseudomonas aeruginosa PAK polar pili. Infect Immun. 1983 Oct;42(1):113–121. doi: 10.1128/iai.42.1.113-121.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Worobec E. A., Paranchych W., Parker J. M., Taneja A. K., Hodges R. S. Antigen-antibody interaction. The immunodominant region of EDP208 pili. J Biol Chem. 1985 Jan 25;260(2):938–943. [PubMed] [Google Scholar]
  32. Yoshitake S., Yamada Y., Ishikawa E., Masseyeff R. Conjugation of glucose oxidase from Aspergillus niger and rabbit antibodies using N-hydroxysuccinimide ester of N-(4-carboxycyclohexylmethyl)-maleimide. Eur J Biochem. 1979 Nov;101(2):395–399. doi: 10.1111/j.1432-1033.1979.tb19731.x. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES