title: Purification and preliminary characterization of the zonula occludens toxin receptor from human (CaCo2) and murine (IEC6) intestinal cell lines.
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.
authors.affiliation: Division of Pediatric Gastroenterology, Center for Vaccine Development, University of Maryland, School of Medicine, Baltimore, MD 21201, USA. uzzau@ssmain.uniss.it
authors.forename: S
authors.initials: S
authors.lastname: Uzzau
authors.forename: R
authors.initials: R
authors.lastname: Lu
authors.forename: W
authors.initials: W
authors.lastname: Wang
authors.forename: C
authors.initials: C
authors.lastname: Fiore
authors.forename: A
authors.initials: A
authors.lastname: Fasano
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D018938:Caco-2 Cells; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D002417:Cattle; D002460:Cell Line; D020848:Chimerin 1 / Q000235:genetics; D002772:Cholera Toxin / Q000378:metabolism*; D004285:Dogs; D004731:Endotoxins; D004847:Epithelial Cells / Q000378:metabolism; D017353:Gene Deletion; D006801:Humans; D015151:Immunoblotting; D007413:Intestinal Mucosa / Q000166:cytology* / Q000378:metabolism; D058619:Maltose-Binding Proteins; D008969:Molecular Sequence Data; D051381:Rats; D011956:Receptors, Cell Surface / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014734:Vibrio cholerae / Q000378:metabolism
chemical_list: D002352:Carrier Proteins; D020848:Chimerin 1; D004731:Endotoxins; D058619:Maltose-Binding Proteins; D011956:Receptors, Cell Surface; D011993:Recombinant Fusion Proteins; C402827:zonula occludens toxin receptor; C077728:zonula occludens toxin, Vibrio cholerae; D002772:Cholera Toxin