Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1995 Feb;63(2):640–646. doi: 10.1128/iai.63.2.640-646.1995

Binding of the fibrillar CS3 adhesin of enterotoxigenic Escherichia coli to rabbit intestinal glycoproteins is competitively prevented by GalNAc beta 1-4Gal-containing glycoconjugates.

C Wennerås 1, J R Neeser 1, A M Svennerholm 1
PMCID: PMC173043  PMID: 7822033

Abstract

We have attempted to characterize the binding specificity of the coli surface 3 (CS3) subcomponent of colonization factor antigen II of enterotoxigenic Escherichia coli, by means of an immunoblot method in which the binding of fimbriated bacteria to sodium dodecyl sulfate-polyacrylamide gel electrophoresis-separated rabbit intestinal cell membranes was evaluated. Isolated CS3 fibrillae as well as bacteria expressing CS3 on their surface bound to several intestinal cell membrane structures, i.e., structures present in the electrophoretic front and in the 30- to 35-kDa range and, most prominently, 120- to 140-kDa structures. Delipidization and protein digestion of the rabbit brush borders revealed that CS3 bound to structures of a proteinaceous nature. Sodium meta-periodate oxidation of the intestinal cell membranes abolished all their CS3 binding activity, indicating that CS3 bound to carbohydrate moieties of glycoproteins. The binding of CS3 to the separated intestinal proteins could also be inhibited by preincubation with the lectin derived from Maackia amurensis, indicating that CS3 bound to galactoproteins in the rabbit intestine. Inhibition experiments using equimolar amounts of various gangliosides demonstrated that GM1, asialo-GM1, and GM2 inhibited the binding of CS3 equally well, whereas GM3 was not as effective. These results suggested that the critical CS3 binding epitope consisted of the carbohydrate sequence GalNAc beta 1-4Gal. This was supported by electron microscopic experiments showing that this disaccharide, O linked to bovine serum albumin via a spacer, localized around CS3-positive bacteria but not at all around corresponding CS3-negative mutants. Furthermore, CS3-expressing bacteria recognized this neoglycoprotein when it was immobilized on nitrocellulose. The GalNAc beta 1-4Gal disaccharide has also been implicated as a binding structure for other pathogenic bacteria such as enteropathogenic E. coli and Pseudomonas aeruginosa.

Full Text

The Full Text of this article is available as a PDF (436.6 KB).

Selected References

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

  1. Black R. E. Epidemiology of diarrhoeal disease: implications for control by vaccines. Vaccine. 1993;11(2):100–106. doi: 10.1016/0264-410x(93)90002-f. [DOI] [PubMed] [Google Scholar]
  2. Blanchard D., Cartron J. P., Fournet B., Montreuil J., van Halbeek H., Vliegenthart J. F. Primary structure of the oligosaccharide determinant of blood group Cad specificity. J Biol Chem. 1983 Jun 25;258(12):7691–7695. [PubMed] [Google Scholar]
  3. Bühler T., Hoschützky H., Jann K. Analysis of colonization factor antigen I, an adhesin of enterotoxigenic Escherichia coli O78:H11: fimbrial morphology and location of the receptor-binding site. Infect Immun. 1991 Nov;59(11):3876–3882. doi: 10.1128/iai.59.11.3876-3882.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Donald A. S., Soh C. P., Watkins W. M., Morgan W. T. N-Acetyl-D-galactosaminyl-beta-(1 goes to 4)-d-galactose: a terminal non-reducing structure in human blood group Sda-active Tamm-Horsfall urinary glycoprotein. Biochem Biophys Res Commun. 1982 Jan 15;104(1):58–65. doi: 10.1016/0006-291x(82)91940-4. [DOI] [PubMed] [Google Scholar]
  5. Evans D. G., Evans D. J., Jr, Clegg S., Pauley J. A. Purification and characterization of the CFA/I antigen of enterotoxigenic Escherichia coli. Infect Immun. 1979 Aug;25(2):738–748. doi: 10.1128/iai.25.2.738-748.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Evans D. G., Silver R. P., Evans D. J., Jr, Chase D. G., Gorbach S. L. Plasmid-controlled colonization factor associated with virulence in Esherichia coli enterotoxigenic for humans. Infect Immun. 1975 Sep;12(3):656–667. doi: 10.1128/iai.12.3.656-667.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Jagannatha H. M., Sharma U. K., Ramaseshan T., Surolia A., Balganesh T. S. Identification of carbohydrate structures as receptors for localised adherent enteropathogenic Escherichia coli. Microb Pathog. 1991 Oct;11(4):259–268. doi: 10.1016/0882-4010(91)90030-e. [DOI] [PubMed] [Google Scholar]
  8. Jonson G., Sanchez J., Svennerholm A. M. Expression and detection of different biotype-associated cell-bound haemagglutinins of Vibrio cholerae O1. J Gen Microbiol. 1989 Jan;135(1):111–120. doi: 10.1099/00221287-135-1-111. [DOI] [PubMed] [Google Scholar]
  9. Karlsson K. A. Animal glycosphingolipids as membrane attachment sites for bacteria. Annu Rev Biochem. 1989;58:309–350. doi: 10.1146/annurev.bi.58.070189.001521. [DOI] [PubMed] [Google Scholar]
  10. Knutton S., Lloyd D. R., Candy D. C., McNeish A. S. Adhesion of enterotoxigenic Escherichia coli to human small intestinal enterocytes. Infect Immun. 1985 Jun;48(3):824–831. doi: 10.1128/iai.48.3.824-831.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Krivan H. C., Roberts D. D., Ginsburg V. Many pulmonary pathogenic bacteria bind specifically to the carbohydrate sequence GalNAc beta 1-4Gal found in some glycolipids. Proc Natl Acad Sci U S A. 1988 Aug;85(16):6157–6161. doi: 10.1073/pnas.85.16.6157. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Levine M. M., Ristaino P., Marley G., Smyth C., Knutton S., Boedeker E., Black R., Young C., Clements M. L., Cheney C. Coli surface antigens 1 and 3 of colonization factor antigen II-positive enterotoxigenic Escherichia coli: morphology, purification, and immune responses in humans. Infect Immun. 1984 May;44(2):409–420. doi: 10.1128/iai.44.2.409-420.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lindahl M., Faris A., Wadstrom T. Colonization factor antigen on enterotoxigenic Escherichia coli is a sialic-specific lectin. Lancet. 1982 Jul 31;2(8292):280–280. doi: 10.1016/s0140-6736(82)90368-3. [DOI] [PubMed] [Google Scholar]
  15. Lopez-Vidal Y., Svennerholm A. M. Monoclonal antibodies against the different subcomponents of colonization factor antigen II of enterotoxigenic Escherichia coli. J Clin Microbiol. 1990 Sep;28(9):1906–1912. doi: 10.1128/jcm.28.9.1906-1912.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. MILLER D., CRANE R. K. A procedure for the isolation of the epithelial brush border membrane of hamster small intestine. Anal Biochem. 1961 Jun;2:284–286. doi: 10.1016/s0003-2697(61)80014-6. [DOI] [PubMed] [Google Scholar]
  17. Moon H. W., Bunn T. O. Vaccines for preventing enterotoxigenic Escherichia coli infections in farm animals. Vaccine. 1993;11(2):213–200. doi: 10.1016/0264-410X(93)90020-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Neeser J. R., Chambaz A., Del Vedovo S., Prigent M. J., Guggenheim B. Specific and nonspecific inhibition of adhesion of oral actinomyces and streptococci to erythrocytes and polystyrene by caseinoglycopeptide derivatives. Infect Immun. 1988 Dec;56(12):3201–3208. doi: 10.1128/iai.56.12.3201-3208.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Neeser J. R., Chambaz A., Golliard M., Link-Amster H., Fryder V., Kolodziejczyk E. Adhesion of colonization factor antigen II-positive enterotoxigenic Escherichia coli strains to human enterocytelike differentiated HT-29 cells: a basis for host-pathogen interactions in the gut. Infect Immun. 1989 Dec;57(12):3727–3734. doi: 10.1128/iai.57.12.3727-3734.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Orø H. S., Kolstø A. B., Wennerås C., Svennerholm A. M. Identification of asialo GM1 as a binding structure for Escherichia coli colonization factor antigens. FEMS Microbiol Lett. 1990 Nov;60(3):289–292. doi: 10.1016/0378-1097(90)90319-l. [DOI] [PubMed] [Google Scholar]
  21. Pieroni P., Worobec E. A., Paranchych W., Armstrong G. D. Identification of a human erythrocyte receptor for colonization factor antigen I pili expressed by H10407 enterotoxigenic Escherichia coli. Infect Immun. 1988 May;56(5):1334–1340. doi: 10.1128/iai.56.5.1334-1340.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Podolsky D. K. Oligosaccharide structures of human colonic mucin. J Biol Chem. 1985 Jul 15;260(14):8262–8271. [PubMed] [Google Scholar]
  23. Rauvala H., Finne J. Structural similarity of the terminal carbohydrate sequences of glycoproteins and glycolipids. FEBS Lett. 1979 Jan 1;97(1):1–8. doi: 10.1016/0014-5793(79)80039-3. [DOI] [PubMed] [Google Scholar]
  24. Svennerholm A. M., Wennerås C., Holmgren J., McConnell M. M., Rowe B. Roles of different coli surface antigens of colonization factor antigen II in colonization by and protective immunogenicity of enterotoxigenic Escherichia coli in rabbits. Infect Immun. 1990 Feb;58(2):341–346. doi: 10.1128/iai.58.2.341-346.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Svennerholm L., Fredman P. A procedure for the quantitative isolation of brain gangliosides. Biochim Biophys Acta. 1980 Jan 18;617(1):97–109. doi: 10.1016/0005-2760(80)90227-1. [DOI] [PubMed] [Google Scholar]
  26. Thomas L. V., McConnell M. M., Rowe B., Field A. M. The possession of three novel coli surface antigens by enterotoxigenic Escherichia coli strains positive for the putative colonization factor PCF8775. J Gen Microbiol. 1985 Sep;131(9):2319–2326. doi: 10.1099/00221287-131-9-2319. [DOI] [PubMed] [Google Scholar]
  27. 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]
  28. Wang W. C., Cummings R. D. The immobilized leukoagglutinin from the seeds of Maackia amurensis binds with high affinity to complex-type Asn-linked oligosaccharides containing terminal sialic acid-linked alpha-2,3 to penultimate galactose residues. J Biol Chem. 1988 Apr 5;263(10):4576–4585. [PubMed] [Google Scholar]
  29. Wennerås C., Holmgren J., Svennerholm A. M. The binding of colonization factor antigens of enterotoxigenic Escherichia coli to intestinal cell membrane proteins. FEMS Microbiol Lett. 1990 Jan 1;54(1-3):107–112. doi: 10.1016/0378-1097(90)90266-s. [DOI] [PubMed] [Google Scholar]

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

RESOURCES