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Infection and Immunity logoLink to Infection and Immunity
. 1986 Apr;52(1):18–25. doi: 10.1128/iai.52.1.18-25.1986

Identification and characterization of mouse small intestine mucosal receptors for Escherichia coli K-12(K88ab).

D C Laux, E F McSweegan, T J Williams, E A Wadolkowski, P S Cohen
PMCID: PMC262191  PMID: 3007359

Abstract

Adhesion of 3H-labeled Escherichia coli K-12(K88ab) to CD-1 mouse small intestine mucus and brush border preparations, immobilized on polystyrene, was studied. E. coli K12(K88ab) was shown to adhere readily to either crude mucus or brush border preparations, but not to bovine serum albumin. In contrast, the nearly isogenic E. coli K-12 strain, i.e., lacking the K88ab plasmid, did not bind well to either mucus, brush borders, or bovine serum albumin. The adhesion of E. coli K-12(K88ab) to both mucus and brush borders required pilus expression (i.e., growth at temperatures greater than 18 degrees C) and was inhibited by pretreatment of either mucus or brush borders with trypsin, pronase, or sodium metaperiodate and by the presence of D-galactosamine. Crude mucus was fractionated by gel filtration, and the proteins in receptor-containing fractions were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Separated proteins were Western blotted to nitrocellulose. Adhesion of 35SO4-labeled E. coli K-12(K88ab) and 35SO4-labeled E. coli K-12 to Western blots followed by autoradiography revealed two E. coli K-12(K88ab)-specific mucus receptor proteins (57 and 64 kilodaltons). Brush borders contained the same two receptor proteins present in mucus and an additional 91-kilodalton receptor protein.

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Selected References

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  1. Allen A., Snary D. The structure and function of gastric mucus. Gut. 1972 Aug;13(8):666–672. doi: 10.1136/gut.13.8.666. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Anderson M. J., Whitehead J. S., Kim Y. S. Interaction of Escherichia coli K88 antigen with porcine intestinal brush border membranes. Infect Immun. 1980 Sep;29(3):897–901. doi: 10.1128/iai.29.3.897-901.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bijlsma I. G., de Nijs A., van der Meer C., Frik J. F. Different pig phenotypes affect adherence of Escherichia coli to jejunal brush borders by K88ab, K88ac, or K88ad antigen. Infect Immun. 1982 Sep;37(3):891–894. doi: 10.1128/iai.37.3.891-894.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cohen P. S., Arruda J. C., Williams T. J., Laux D. C. Adhesion of a human fecal Escherichia coli strain to mouse colonic mucus. Infect Immun. 1985 Apr;48(1):139–145. doi: 10.1128/iai.48.1.139-145.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. DISCHE Z. New color reactions for determination of sugars in polysaccharides. Methods Biochem Anal. 1955;2:313–358. doi: 10.1002/9780470110188.ch11. [DOI] [PubMed] [Google Scholar]
  6. Etzler M. E. Lectins as probes in studies of intestinal glycoproteins and glycolipids. Am J Clin Nutr. 1979 Jan;32(1):133–138. doi: 10.1093/ajcn/32.1.133. [DOI] [PubMed] [Google Scholar]
  7. Forstner J. F. Intestinal mucins in health and disease. Digestion. 1978;17(3):234–263. doi: 10.1159/000198115. [DOI] [PubMed] [Google Scholar]
  8. Forstner J., Taichman N., Kalnins V., Forstner G. Intestinal goblet cell mucus: isolation and identification by immunofluorescence of a goblet cell glycoprotein. J Cell Sci. 1973 Mar;12(2):585–602. doi: 10.1242/jcs.12.2.585. [DOI] [PubMed] [Google Scholar]
  9. Frantz J. C., Jaso-Friedman L., Robertson D. C. Binding of Escherichia coli heat-stable enterotoxin to rat intestinal cells and brush border membranes. Infect Immun. 1984 Feb;43(2):622–630. doi: 10.1128/iai.43.2.622-630.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gibbons R. A., Jones G. W., Sellwood R. An attempt to identify the intestinal receptor for the K88 adhesin by means of a haemagglutination inhibition test using glycoproteins and fractions from sow colostrum. J Gen Microbiol. 1975 Feb;86(2):228–240. doi: 10.1099/00221287-86-2-228. [DOI] [PubMed] [Google Scholar]
  11. Glossmann H., Neville D. M., Jr Glycoproteins of cell surfaces. A comparative study of three different cell surfaces of the rat. J Biol Chem. 1971 Oct 25;246(20):6339–6346. [PubMed] [Google Scholar]
  12. Guinée P. A., Jansen W. H. Behavior of Escherichia coli K antigens K88ab, K88ac, and K88ad in immunoelectrophoresis, double diffusion, and hemagglutination. Infect Immun. 1979 Mar;23(3):700–705. doi: 10.1128/iai.23.3.700-705.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Izhar M., Nuchamowitz Y., Mirelman D. Adherence of Shigella flexneri to guinea pig intestinal cells is mediated by a mucosal adhesion. Infect Immun. 1982 Mar;35(3):1110–1118. doi: 10.1128/iai.35.3.1110-1118.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Jones G. W., Rutter J. M. Role of the K88 antigen in the pathogenesis of neonatal diarrhea caused by Escherichia coli in piglets. Infect Immun. 1972 Dec;6(6):918–927. doi: 10.1128/iai.6.6.918-927.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Laskey R. A., Mills A. D. Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. Eur J Biochem. 1975 Aug 15;56(2):335–341. doi: 10.1111/j.1432-1033.1975.tb02238.x. [DOI] [PubMed] [Google Scholar]
  17. Lynch K. R., Pennica D., Ennis H. L., Cohen P. S. Separation and purification of the mRNAs for vesicular stomatitis virus NS and M proteins. Virology. 1979 Oct 15;98(1):251–254. doi: 10.1016/0042-6822(79)90543-9. [DOI] [PubMed] [Google Scholar]
  18. Mooi F. R., de Graaf F. K., van Embden J. D. Cloning, mapping and expression of the genetic determinant that encodes for the K88ab antigen. Nucleic Acids Res. 1979 Mar;6(3):849–865. doi: 10.1093/nar/6.3.849. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. O'Hanley P., Lark D., Normark S., Falkow S., Schoolnik G. K. Mannose-sensitive and Gal-Gal binding Escherichia coli pili from recombinant strains. Chemical, functional, and serological properties. J Exp Med. 1983 Nov 1;158(5):1713–1719. doi: 10.1084/jem.158.5.1713. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. ORSKOV I., ORSKOV F., SOJKA W. J., LEACH J. M. Simultaneous occurrence of E. coli B and Lantigens in strains from diseased swine. Influence of cultivation temperature. Two new E. coli Kantigens: K 87 and K 88. Acta Pathol Microbiol Scand. 1961;53:404–422. [PubMed] [Google Scholar]
  21. ORSKOV I., ORSKOV F., SOJKA W. J., WITTIG W. K ANTIGENS K88AB(L) AND K88AC(L) IN E. COLI. A NEW O ANTIGEN: 0147 AND A NEW K ANTIGEN: K89(B). Acta Pathol Microbiol Scand. 1964;62:439–447. doi: 10.1111/apm.1964.62.3.439. [DOI] [PubMed] [Google Scholar]
  22. Orskov I., Orskov F. Episome-carried surface antigen K88 of Escherichia coli. I. Transmission of the determinant of the K88 antigen and influence on the transfer of chromosomal markers. J Bacteriol. 1966 Jan;91(1):69–75. doi: 10.1128/jb.91.1.69-75.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Parry S. H., Porter P. Immunological aspects of cell membrane adhesion demonstrated by porcine enteropathogenic Escherichia coli. Immunology. 1978 Jan;34(1):41–49. [PMC free article] [PubMed] [Google Scholar]
  24. Porter W. H. Application of nitrous acid deamination of hexosamines to the simultaneous GLC determination of neutral and amino sugars in glycoproteins. Anal Biochem. 1975 Jan;63(1):27–43. doi: 10.1016/0003-2697(75)90186-4. [DOI] [PubMed] [Google Scholar]
  25. Runnels P. L., Moon H. W., Schneider R. A. Development of resistance with host age to adhesion of K99+ Escherichia coli to isolated intestinal epithelial cells. Infect Immun. 1980 Apr;28(1):298–300. doi: 10.1128/iai.28.1.298-300.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. SVENNERHOLM L. Quantitative estimation of sialic acids. II. A colorimetric resorcinol-hydrochloric acid method. Biochim Biophys Acta. 1957 Jun;24(3):604–611. doi: 10.1016/0006-3002(57)90254-8. [DOI] [PubMed] [Google Scholar]
  27. 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]
  28. Sellwood R., Gibbons R. A., Jones G. W., Rutter J. M. Adhesion of enteropathogenic Escherichia coli to pig intestinal brush borders: the existence of two pig phenotypes. J Med Microbiol. 1975 Aug;8(3):405–411. doi: 10.1099/00222615-8-3-405. [DOI] [PubMed] [Google Scholar]
  29. Smit H., Gaastra W., Kamerling J. P., Vliegenthart J. F., de Graaf F. K. Isolation and structural characterization of the equine erythrocyte receptor for enterotoxigenic Escherichia coli K99 fimbrial adhesin. Infect Immun. 1984 Nov;46(2):578–584. doi: 10.1128/iai.46.2.578-584.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Smith H. W., Huggins M. B. The influence of plasmid-determined and other characteristics of enteropathogenic Escherichia coli on their ability to proliferate in the alimentary tracts of piglets, calves and lambs. J Med Microbiol. 1978 Nov;11(4):471–492. doi: 10.1099/00222615-11-4-471. [DOI] [PubMed] [Google Scholar]
  31. 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]
  32. Stirm S., Orskov F., Orskov I., Birch-Andersen A. Episome-carried surface antigen K88 of Escherichia coli. 3. Morphology. J Bacteriol. 1967 Feb;93(2):740–748. doi: 10.1128/jb.93.2.740-748.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. 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]
  34. Weiser M. M. Intestinal epithelial cell surface membrane glycoprotein synthesis. I. An indicator of cellular differentiation. J Biol Chem. 1973 Apr 10;248(7):2536–2541. [PubMed] [Google Scholar]

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