Characterization of porcine intestinal receptors for the K88ac fimbrial adhesin of Escherichia coli as mucin-type sialoglycoproteins

Infect Immun. 1994 Dec;62(12):5404-10. doi: 10.1128/iai.62.12.5404-5410.1994.

Abstract

We have previously identified two K88ac adhesion receptors (210 and 240 kDa) which are present in membrane preparations from adhesive but not nonadhesive porcine intestinal brush border cells; these adhesin receptors are postulated to be important determinants of the susceptibility of pigs to K88ac+ enterotoxigenic Escherichia coli infections (A.K. Erickson, J.A. Willgohs, S.Y. McFarland, D.A. Benfield, and D.F. Francis, Infect. Immun. 60:983-988, 1992). We now describe a procedure for the purification of these two receptors. Receptors were solubilized from adhesive intestinal brush border vesicles using deoxycholate and were purified by gel filtration chromatography on Sepharose CL-4B and then by hydroxyapatite chromatography. Amino acid compositional analyses indicated that the two receptors have similar amino acid compositions. The most distinguishing characteristic of both receptors is a high percentage of threonine and proline residues. Neuraminidase treatment caused the K88ac adhesin receptors to migrate with a slower mobility on sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels, indicating that these receptors are sialoglycoproteins. Results from lectin-binding studies indicated that the receptors contain O-linked oligosaccharides composed of galactosyl (beta-1,3)N-acetylgalactosamine, alpha-linked fucose, galactosyl(beta-1,4)N-acetylglucosamine, sialic acid, galactose, and N-acetylgalactosamine. Collectively, these characteristics indicate that the K88ac adhesin receptors are mucin-type sialoglycoproteins.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Amino Acids / analysis
  • Animals
  • Antigens, Bacterial*
  • Antigens, Surface / immunology
  • Bacterial Adhesion / physiology
  • Carbohydrate Sequence
  • Carbohydrates / chemistry
  • Escherichia coli / immunology*
  • Escherichia coli Proteins*
  • Fimbriae Proteins*
  • Fimbriae, Bacterial / immunology
  • Intestines / chemistry*
  • Lectins / metabolism
  • Microvilli / chemistry
  • Molecular Sequence Data
  • Mucins / chemistry*
  • Receptors, Immunologic / chemistry*
  • Receptors, Immunologic / isolation & purification
  • Sialoglycoproteins / chemistry*
  • Sialoglycoproteins / classification
  • Swine

Substances

  • Amino Acids
  • Antigens, Bacterial
  • Antigens, Surface
  • Carbohydrates
  • Escherichia coli Proteins
  • K88 antigen, E coli
  • Lectins
  • Mucins
  • Receptors, Immunologic
  • Sialoglycoproteins
  • bacterial adhesin receptor
  • Fimbriae Proteins