Skip to main content
Biochemical Journal logoLink to Biochemical Journal
. 1988 Oct 1;255(1):105–111. doi: 10.1042/bj2550105

Purification and characterization of CS2, a sialic acid-specific haemagglutinin of enterotoxigenic Escherichia coli.

P O Sjöberg 1, M Lindahl 1, J Porath 1, T Wadström 1
PMCID: PMC1135196  PMID: 2904260

Abstract

CS2 fimbriae of enterotoxigenic Escherichia coli were purified and characterized. The surface haemagglutinins (fimbriae) were detached by sonication from a strain producing only the CS2 fimbriae. Isolation was carried out by gel filtration on a Sepharose 4B column. After depolymerization, the fimbriae subunits were purified on a Sephacryl S-300 column in 8.0 M-guanidinium chloride. From 1 litre of medium, 4-6 mg of purified fimbriae was obtained. We found that CS2 fimbriae were completely dissociated by saturated guanidinium chloride into subunits with a molecular mass of 16.5 kDa. CS2 fimbriae was sialic acid-specific, since sialic acids were the most potent inhibitors, and neuraminidase treatment of erythrocytes abolished haemagglutination. Both fimbriae and fimbrial subunits were found to bind to bovine erythrocytes. The binding of subunits to erythrocytes could be inhibited with low concentrations of sialyl-lactose.

Full text

PDF
105

Images in this article

Selected References

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

  1. Blobel G., Dobberstein B. Transfer of proteins across membranes. I. Presence of proteolytically processed and unprocessed nascent immunoglobulin light chains on membrane-bound ribosomes of murine myeloma. J Cell Biol. 1975 Dec;67(3):835–851. doi: 10.1083/jcb.67.3.835. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. Bäck E., Möllby R., Kaijser B., Stintzing G., Wadström T., Habte D. Enterotoxigenic Escherichia coli and other gram-negative bacteria of infantile diarrhea: surface antigens, hemagglutinins, colonization factor antigen, and loss of enterotoxigenicity. J Infect Dis. 1980 Sep;142(3):318–327. doi: 10.1093/infdis/142.3.318. [DOI] [PubMed] [Google Scholar]
  4. DUGUID J. P., SMITH I. W., DEMPSTER G., EDMUNDS P. N. Non-flagellar filamentous appendages (fimbriae) and haemagglutinating activity in Bacterium coli. J Pathol Bacteriol. 1955 Oct;70(2):335–348. doi: 10.1002/path.1700700210. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. 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]
  7. Evans D. G., Evans D. J., Jr, Tjoa W. Hemagglutination of human group A erythrocytes by enterotoxigenic Escherichia coli isolated from adults with diarrhea: correlation with colonization factor. Infect Immun. 1977 Nov;18(2):330–337. doi: 10.1128/iai.18.2.330-337.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Faris A., Ljungh A., Wadström T. Rapid identification of colonization factor antigens (CFA/I and CFA/II) of enterotoxigenic E. coli (ETEC) and O-antigens of enteropathogenic E. coli (EPEC) by coagglutination test. Zentralbl Bakteriol Mikrobiol Hyg A. 1985 Jul;259(4):477–484. doi: 10.1016/s0176-6724(85)80079-1. [DOI] [PubMed] [Google Scholar]
  9. Faris A., Sellei J., Lindahl M., Wadström T. Haemagglutination of bovine erythrocytes by enterotoxigenic E. coli (ETEC) of O 6 serogroup: evidence for glycoconjugate receptor heterogeneity. Zentralbl Bakteriol Mikrobiol Hyg A. 1982 Nov;253(2):175–182. [PubMed] [Google Scholar]
  10. Firon N., Ofek I., Sharon N. Carbohydrate specificity of the surface lectins of Escherichia coli, Klebsiella pneumoniae, and Salmonella typhimurium. Carbohydr Res. 1983 Aug 16;120:235–249. doi: 10.1016/0008-6215(83)88019-7. [DOI] [PubMed] [Google Scholar]
  11. Guerrant R. L. Microbial toxins and diarrhoeal diseases: introduction and overview. Ciba Found Symp. 1985;112:1–13. doi: 10.1002/9780470720936.ch1. [DOI] [PubMed] [Google Scholar]
  12. Jacobs A. A., van Mechelen J. R., de Graaf F. K. Effect of chemical modifications on the K99 and K88ab fibrillar adhesins of Escherichia coli. Biochim Biophys Acta. 1985 Nov 29;832(2):148–155. doi: 10.1016/0167-4838(85)90326-7. [DOI] [PubMed] [Google Scholar]
  13. Korhonen T. K., Väisänen V., Saxén H., Hultberg H., Svenson S. B. P-antigen-recognizing fimbriae from human uropathogenic Escherichia coli strains. Infect Immun. 1982 Jul;37(1):286–291. doi: 10.1128/iai.37.1.286-291.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  15. March S. C., Parikh I., Cuatrecasas P. A simplified method for cyanogen bromide activation of agarose for affinity chromatography. Anal Biochem. 1974 Jul;60(1):149–152. doi: 10.1016/0003-2697(74)90139-0. [DOI] [PubMed] [Google Scholar]
  16. McMichael J. C., Ou J. T. Structure of common pili from Escherichia coli. J Bacteriol. 1979 Jun;138(3):969–975. doi: 10.1128/jb.138.3.969-975.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Mooi F. R., Wijfjes A., de Graaf F. K. Identification and characterization of precursors in the biosynthesis of the K88ab fimbria of Escherichia coli. J Bacteriol. 1983 Apr;154(1):41–49. doi: 10.1128/jb.154.1.41-49.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. Parkkinen J., Finne J., Achtman M., Väisänen V., Korhonen T. K. Escherichia coli strains binding neuraminyl alpha 2-3 galactosides. Biochem Biophys Res Commun. 1983 Mar 16;111(2):456–461. doi: 10.1016/0006-291x(83)90328-5. [DOI] [PubMed] [Google Scholar]
  20. Smith H. W., Linggood M. A. Observations on the pathogenic properties of the K88, Hly and Ent plasmids of Escherichia coli with particular reference to porcine diarrhoea. J Med Microbiol. 1971 Nov;4(4):467–485. doi: 10.1099/00222615-4-4-467. [DOI] [PubMed] [Google Scholar]
  21. Smyth C. J. Two mannose-resistant haemagglutinins on enterotoxigenic Escherichia coli of serotype O6:K15:H16 or H-isolated from travellers' and infantile diarrhoea. J Gen Microbiol. 1982 Sep;128(9):2081–2096. doi: 10.1099/00221287-128-9-2081. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES