globotriaosylceramide and Dysentery--Bacillary

Shiga toxins consist of an A-moiety and five B-moieties able to bind the neutral glycosphingolipid globotriaosylceramide (Gb3) on the cell surface. To intoxicate cells efficiently, the toxin A-moiety has to be cleaved by furin and transported retrogradely to the Golgi apparatus and to the endoplasmic reticulum. The enzymatically active part of the A-moiety is then translocated to the cytosol, where it inhibits protein synthesis and in some cell types induces apoptosis. Protection of cells can be provided either by inhibiting binding of the toxin to cells or by interfering with any of the subsequent steps required for its toxic effect. In this article we provide a brief overview of the interaction of Shiga toxins with cells, describe some compounds and conditions found to protect cells against Shiga toxins, and discuss whether they might also provide protection in animals and humans.

Topics: Animals; Antidotes; Apoptosis; Bacterial Proteins; Dysentery, Bacillary; Hemolytic-Uremic Syndrome; Host-Pathogen Interactions; Humans; Protein Biosynthesis; Protein Conformation; Protein Transport; Shiga Toxins; Shiga-Toxigenic Escherichia coli; Shigella dysenteriae; Structure-Activity Relationship; Trihexosylceramides

Topics: Bacterial Toxins; Base Sequence; Carbohydrate Sequence; Disease Models, Animal; Dysentery, Bacillary; Endocytosis; Enterotoxins; Escherichia coli; Gastrointestinal Hemorrhage; Gene Expression Regulation, Bacterial; Genes, Bacterial; Hemolytic-Uremic Syndrome; Humans; Molecular Sequence Data; Protein Biosynthesis; Receptors, Cell Surface; Shiga Toxin 1; Shiga Toxin 2; Shiga Toxins; Shigella; Structure-Activity Relationship; Trihexosylceramides; Virulence

Biomembranes are hard to compress laterally, and membrane area compressibility has not been associated with biological processes. Using X-ray surface scattering, we observed that bacterial Shiga toxin compresses lipid packing in a gel phase monolayer upon binding to its cellular receptor, the glycolipid Gb3. This toxin-induced reorganization of lipid packing reached beyond the immediate membrane patch that the protein was bound to, and linkers separating the Gb3 carbohydrate and ceramide moieties modulated the toxin's capacity to compress the membrane. Within a natural membrane, asymmetric compression of the toxin-bound leaflet could provide a mechanism to initiate narrow membrane bending, as observed upon toxin entry into cells. Such lipid compression and long-range membrane reorganization by glycolipid-binding proteins represent novel concepts in membrane biology that have direct implications for the construction of endocytic pits in clathrin-independent endocytosis.

Topics: Cell Membrane; Dysentery, Bacillary; Endocytosis; Humans; Models, Molecular; Phosphatidylethanolamines; Shiga Toxin; Shigella dysenteriae; Trihexosylceramides

Lateral variation of the in-plane orientation of lipids in a bilayer is referred to as texture. The influence of the protein Shiga toxin on orientational membrane texture was studied in phosphatidylcholine lipid bilayers using polarization two-photon fluorescence microscopy and atomic force microscopy. A content of 1% of glycosphingolipid globotriaosylceramide (Gb3) receptor lipids in a bilayer was used to bind the Shiga toxin B-subunit to the surface of gel domains. Binding of the Shiga toxin B-subunit to lipids led to the modulation of orientational membrane texture in gel domains and induced membrane reordering. When Shiga toxin was added above the lipid chain melting temperature, the toxin interaction with the membrane induced rearrangement and clustering of Gb3 lipids that resulted in the long range order and alignment of lipids in gel domains. The toxin induced redistribution of Gb3 lipids inside gel domains is governed by the temperature at which Shiga toxin was added to the membrane: above or below the phase transition. The temperature is thus one of the critical factors controlling lipid organization and texture in the presence of Shiga toxin. Lipid chain ordering imposed by Shiga toxin binding can be another factor driving the reconstruction of lipid organization and crystallization of lipids inside gel domains.

Topics: Dysentery, Bacillary; Humans; Lipid Bilayers; Phase Transition; Phospholipids; Shiga Toxin; Shigella dysenteriae; Trihexosylceramides

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.

Topics: Animals; Bacterial Toxins; Binding, Competitive; Chromatography, High Pressure Liquid; Dysentery, Bacillary; Globosides; Glycosphingolipids; HeLa Cells; Humans; Jejunum; Liposomes; Oligosaccharides; Rabbits; Receptors, Cell Surface; Receptors, Immunologic; Shiga Toxins; Shigella dysenteriae; Trihexosylceramides