Pathogenesis of shigella diarrhea. XI. Isolation of a shigella toxin-binding glycolipid from rabbit jejunum and HeLa cells and its identification as globotriaosylceramide.
A glycolipid that specifically binds shigella toxin was isolated from both HeLa cells and rabbit jejunal mucosa and identified as globotriaosylceramide (Gb3) by its identical mobility on HPTLC to authentic erythrocyte Gb3. Toxin also bound to a band tentatively identified as alpha-hydroxylated Gb3. In addition, toxin bound to P1 antigen present in group B human erythrocyte glycolipid extracts. The common feature of the three binding glycolipids is a terminal Gal alpha 1----4Gal disaccharide linked beta 1----4 to either Glc or GlcNAc. Globoisotriaosylceramide, which differs from Gb3 only in possessing a Gal alpha 1----3Gal terminal disaccharide, and LacCer, which lacks the terminal Gal residue of Gb3, were incapable of binding the toxin. Binding was shown to be mediated by the B subunit by the use of isolated toxin A and B subunits and monoclonal subunit-specific antibodies. Gb3-containing liposomes competitively inhibited the binding of toxin to HeLa cell monolayers but did not inhibit toxin-induced cytotoxicity. These studies show an identical carbohydrate-specific glycolipid receptor for shigella toxin in gut and in HeLa cells. The toxin B subunit that mediates this binding has also been shown to recognize a glycoprotein receptor with different sugar specificity. Thus, we have demonstrated that the same small (Mr 6,500) B subunit polypeptide has two distinctive carbohydrate-specific binding sites. The Gal alpha 1----4Gal disaccharide of the glycolipid toxin receptor is also recognized by the Gal-Gal pilus of uropathogenic E. coli. This suggests the possibility that the pilus and toxin B subunit contain homologous sequences. If this is true, it may be possible to use the purified Gal-Gal pilus to produce toxin-neutralizing antibodies.
D000818:Animals; D001427:Bacterial Toxins / Q000378:metabolism*; D001667:Binding, Competitive; D002851:Chromatography, High Pressure Liquid; D004405:Dysentery, Bacillary / Q000209:etiology* / Q000378:metabolism; D005915:Globosides / Q000302:isolation & purification / Q000378:metabolism*; D006028:Glycosphingolipids / Q000378:metabolism*; D006367:HeLa Cells / Q000032:analysis; D006801:Humans; D007583:Jejunum / Q000032:analysis; D008081:Liposomes / Q000378:metabolism; D009844:Oligosaccharides / Q000378:metabolism; D011817:Rabbits; D011956:Receptors, Cell Surface; D011971:Receptors, Immunologic / Q000378:metabolism; D022603:Shiga Toxins; D012762:Shigella dysenteriae; D014281:Trihexosylceramides
D001427:Bacterial Toxins; D005915:Globosides; D006028:Glycosphingolipids; D008081:Liposomes; D009844:Oligosaccharides; D011956:Receptors, Cell Surface; D011971:Receptors, Immunologic; D022603:Shiga Toxins; D014281:Trihexosylceramides; C045390:shigella toxin receptor; C018549:globotriaosylceramide