Enterotoxigenic Escherichia coli TibA glycoprotein adheres to human intestine epithelial cells.
Enterotoxigenic Escherichia coli (ETEC) is capable of invading epithelial cell lines derived from the human ileum and colon. Two separate invasion loci (tia and tib) that direct noninvasive E. coli strains to adhere to and invade cultured human intestine epithelial cells have previously been isolated from the classical ETEC strain H10407. The tib locus directs the synthesis of TibA, a 104-kDa outer membrane glycoprotein. Synthesis of TibA is directly correlated with the adherence and invasion phenotypes of the tib locus, suggesting that this protein is an adhesin and invasin. Here we report the purification of TibA and characterization of its biological activity. TibA was purified by continuous-elution preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Purified TibA was biotin labeled and then shown to bind to HCT8 human ileocecal epithelial cells in a specific and saturable manner. Unlabeled TibA competed with biotin-labeled TibA, suggesting the presence of a specific TibA receptor in HCT8 cells. These results show that TibA acts as an adhesin. Polyclonal anti-TibA antiserum inhibited invasion of ETEC strain H10407 and of recombinant E. coli bearing tib locus clones, suggesting that TibA also acts as an invasin. The ability of TibA to direct epithelial cell adhesion suggests a role for this protein in ETEC pathogenesis.
Lindenthal
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Department of Molecular Biosciences, University of Kansas, Lawrence, Kansas 66045-2106, USA.
Elsinghorst
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EA
D018830:Adhesins, Escherichia coli / Q000302:isolation & purification / Q000502:physiology*; D000818:Animals; D001422:Bacterial Adhesion; D001426:Bacterial Proteins / Q000502:physiology*; D004926:Escherichia coli / Q000502:physiology*; D006023:Glycoproteins / Q000502:physiology; D006801:Humans; D007106:Immune Sera / Q000276:immunology; D007413:Intestinal Mucosa / Q000382:microbiology*; D011817:Rabbits
D018830:Adhesins, Escherichia coli; D001426:Bacterial Proteins; D006023:Glycoproteins; D007106:Immune Sera