Purification and preliminary characterization of the zonula occludens toxin receptor from human (CaCo2) and murine (IEC6) intestinal cell lines

FEMS Microbiol Lett. 2001 Jan 1;194(1):1-5. doi: 10.1111/j.1574-6968.2001.tb09437.x.

Abstract

In the present study, we report the preliminary characterization of the epithelial cell receptor for Vibrio cholerae zonula occludens toxin (Zot). Zot receptor was purified by ligand-affinity chromatography. Analysis of affinity-purified preparations by polyacrylamide gel electrophoresis revealed a protein of ca. 66 kDa. Partial N-terminal sequence obtained from purified murine and human Zot receptor revealed homology between the two proteins and with human alpha-1-chimaerin. Zot protein domain(s) involved in receptor binding were also analyzed by constructing several in frame deletion derivatives of a recombinant fusion Zot protein tagged with maltose binding protein. Our results suggest that Zot binding to its cellular membrane receptor requires a sequence that spans between amino acids 118 and 299.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Caco-2 Cells
  • Carrier Proteins / genetics
  • Carrier Proteins / metabolism
  • Cattle
  • Cell Line
  • Chimerin 1 / genetics
  • Cholera Toxin / metabolism*
  • Dogs
  • Endotoxins
  • Epithelial Cells / metabolism
  • Gene Deletion
  • Humans
  • Immunoblotting
  • Intestinal Mucosa / cytology*
  • Intestinal Mucosa / metabolism
  • Maltose-Binding Proteins
  • Molecular Sequence Data
  • Rats
  • Receptors, Cell Surface / genetics
  • Receptors, Cell Surface / isolation & purification*
  • Receptors, Cell Surface / metabolism
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Vibrio cholerae / metabolism

Substances

  • Carrier Proteins
  • Chimerin 1
  • Endotoxins
  • Maltose-Binding Proteins
  • Receptors, Cell Surface
  • Recombinant Fusion Proteins
  • zonula occludens toxin receptor
  • zonula occludens toxin, Vibrio cholerae
  • Cholera Toxin