Skip to main content
NCBI home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1977 Nov 1;146(5):1169–1181. doi: 10.1084/jem.146.5.1169

Hemagglutination by purified type I Escherichia coli pili

PMCID: PMC2180968  PMID: 336832

Abstract

Many enterobacteria can cause agglutination of erythrocytes, but previous investigations have not proven which components of the bacteria are responsible. We used a strain of Escherichia coli K12 which causes mannose-sensitive hemagglutination (HA) of guinea pig cells. Common pili were purified from these bacteria by shearing them from the bacteria followed by selective precipitation in acid and ammonium sulfate. Isopycnic centrifugation in cesium chloride removed the remaining outer membrane protein contaminants. These pili are pure by electron microscopy and gel electrophoresis. By amino acid analysis, they have a mol wt of 17,099 and consist of 45% nonpolar residues. These purified pili agglutinate guinea pig erythrocytes, a reaction that is inhibited by anti-pili antibodies and by saccharides related in structure to D-mannose. Proteolytic treatment of erythrocytes does not diminish HA but rather increases the pili-induced HA of human cells. Neuraminidase enhances HA and mannosidase slightly diminishes it. It is concluded that purified pili alone cause HA of erythrocytes by binding to mannose-like molecules on the erythrocyte surface. Thus HA by bacterial pili serves as a useful model system for the mechanism of bacterial pili attachment ot cell membranes.

Full Text

The Full Text of this article is available as a PDF (1.6 MB).

Selected References

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

  1. BRINTON C. C., Jr Non-flagellar appendages of bacteria. Nature. 1959 Mar 21;183(4664):782–786. doi: 10.1038/183782a0. [DOI] [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. Buchanan T. M., Pearce W. A. Pili as a mediator of the attachment of gonococci to human erythrocytes. Infect Immun. 1976 May;13(5):1483–1489. doi: 10.1128/iai.13.5.1483-1489.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. DUGUID J. P. Fimbriae and adhesive properties in Klebsiella strains. J Gen Microbiol. 1959 Aug;21:271–286. doi: 10.1099/00221287-21-1-271. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Duguid J. P., Anderson E. S., Campbell I. Fimbriae and adhesive properties in Salmonellae. J Pathol Bacteriol. 1966 Jul;92(1):107–138. doi: 10.1002/path.1700920113. [DOI] [PubMed] [Google Scholar]
  7. Elgsaeter A., Branton D. Intramembrane particle aggregation in erythrocyte ghosts. I. The effects of protein removal. J Cell Biol. 1974 Dec;63(3):1018–1036. doi: 10.1083/jcb.63.3.1018. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fairbanks G., Steck T. L., Wallach D. F. Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry. 1971 Jun 22;10(13):2606–2617. doi: 10.1021/bi00789a030. [DOI] [PubMed] [Google Scholar]
  9. Frye L. D., Edidin M. The rapid intermixing of cell surface antigens after formation of mouse-human heterokaryons. J Cell Sci. 1970 Sep;7(2):319–335. doi: 10.1242/jcs.7.2.319. [DOI] [PubMed] [Google Scholar]
  10. Howard I. K., Sage H. J., Stein M. D., Young N. M., Leon M. A., Dyckes D. F. Studies on a phytohemagglutinin from the lentil. II. Multiple forms of Lens culinaris hemagglutinin. J Biol Chem. 1971 Mar 25;246(6):1590–1595. [PubMed] [Google Scholar]
  11. Kelen A. E., Hathaway A. E., McLeod D. A. Rapid detection of Australia-SH antigen and antibody by a simple and sensitive technique of immunoelectronmicroscopy. Can J Microbiol. 1971 Jul;17(7):993–1000. doi: 10.1139/m71-157. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. 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]
  14. Lewis S. D., Shafer J. A., Goldstein I. J. Kinetic parameters for the binding of p-nitrophenyl alpha-d-mannopyranoside to concanavalin A. Arch Biochem Biophys. 1976 Feb;172(2):689–695. doi: 10.1016/0003-9861(76)90125-9. [DOI] [PubMed] [Google Scholar]
  15. Nicolson G. L. The interactions of lectins with animal cell surfaces. Int Rev Cytol. 1974;39:89–190. doi: 10.1016/s0074-7696(08)60939-0. [DOI] [PubMed] [Google Scholar]
  16. Old D. C. Inhibition of the interaction between fimbrial haemagglutinins and erythrocytes by D-mannose and other carbohydrates. J Gen Microbiol. 1972 Jun;71(1):149–157. doi: 10.1099/00221287-71-1-149. [DOI] [PubMed] [Google Scholar]
  17. Reichstein E., Blostein R. Arrangement of human erythrocyte membrane proteins. J Biol Chem. 1975 Aug 25;250(16):6256–6263. [PubMed] [Google Scholar]
  18. 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]
  19. Schmitges C. J., Henning U. The major proteins of the Escherichia coli outer cell-envelope membrane. Heterogeneity of protein I. Eur J Biochem. 1976 Mar 16;63(1):47–52. doi: 10.1111/j.1432-1033.1976.tb10205.x. [DOI] [PubMed] [Google Scholar]
  20. Schnebli H. P., Roeder C., Tarcsay L. Reaction of lectins with human erythrocytes. III. Surface charge density and agglutination. Exp Cell Res. 1976 Mar 15;98(2):273–276. doi: 10.1016/0014-4827(76)90438-9. [DOI] [PubMed] [Google Scholar]
  21. Simpson R. J., Neuberger M. R., Liu T. Y. Complete amino acid analysis of proteins from a single hydrolysate. J Biol Chem. 1976 Apr 10;251(7):1936–1940. [PubMed] [Google Scholar]
  22. Singer K. A., Morrison M. Effect of metabolic state on agglutination of human erythrocytes by concanavalin A. Biochim Biophys Acta. 1976 Feb 19;426(1):123–131. doi: 10.1016/0005-2736(76)90434-x. [DOI] [PubMed] [Google Scholar]
  23. Tanner M. J., Anstee D. J. A method for the direct demonstration of the lectin-binding components of the human erythrocyte membrane. Biochem J. 1976 Feb 1;153(2):265–270. doi: 10.1042/bj1530265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Vlodavsky I., Inbar M., Sachs L. Temperature-sensitive agglutinability of human erythrocytes by lectins. Biochim Biophys Acta. 1972 Aug 9;274(2):364–369. doi: 10.1016/0005-2736(72)90184-8. [DOI] [PubMed] [Google Scholar]
  25. WARREN L. The thiobarbituric acid assay of sialic acids. J Biol Chem. 1959 Aug;234(8):1971–1975. [PubMed] [Google Scholar]
  26. Waitkins S. A. Fimbrial haemagglutination by Neisseria gonorrhoeae. Br J Vener Dis. 1974 Aug;50(4):272–278. doi: 10.1136/sti.50.4.272. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Yariv J., Kalb A. J., Levitzki A. The interaction of concanavalin A with methyl alpha-D-glucopyranoside. Biochim Biophys Acta. 1968 Sep 3;165(2):303–305. doi: 10.1016/0304-4165(68)90063-9. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES