Purification and preliminary characterization of the zonula occludens toxin receptor from human (CaCo2) and murine (IEC6) intestinal cell lines.
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.
Uzzau
S
S
Division of Pediatric Gastroenterology, Center for Vaccine Development, University of Maryland, School of Medicine, Baltimore, MD 21201, USA. uzzau@ssmain.uniss.it
Lu
R
R
Wang
W
W
Fiore
C
C
Fasano
A
A
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
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