scholarly journals Shiga toxin–binding site for host cell receptor GPP130 reveals unexpected divergence in toxin-trafficking mechanisms

2013 ◽  
Vol 24 (15) ◽  
pp. 2311-2318 ◽  
Author(s):  
Somshuvra Mukhopadhyay ◽  
Brendan Redler ◽  
Adam D. Linstedt
Keyword(s):  
Shiga Toxin ◽  
Transmembrane Domain ◽  
Cell Receptor ◽  
Toxin B ◽  
Toxin Binding ◽  
B Subunit ◽  
Host Cell Receptor ◽  
Developed World ◽  
Trafficking Receptor ◽  
Shed Light

Shiga toxicosis is caused by retrograde trafficking of one of three types of Shiga toxin (STx), STx, STx1, or STx2. Trafficking depends on the toxin B subunits, which for STx and STx1 are identical and bind GPP130, a manganese (Mn)-sensitive intracellular trafficking receptor. Elevated Mn down-regulates GPP130, rendering STx/STx1 harmless. Its effectiveness against STx2, however, which is a serious concern in the developed world, is not known. Here we show that Mn-induced GPP130 down-regulation fails to block STx2 trafficking. To shed light on this result, we tested the purified B subunit of STx2 for binding to GPP130 and found that it failed to interact. We then mapped residues at the interface of the GPP130-STx/STx1 complex. In GPP130, binding mapped to a seven-residue stretch in its lumenal stem domain next to the transmembrane domain. This stretch was required for STx/STx1 transport. In STx/STx1, binding mapped to a histidine–asparagine pair on a surface-exposed loop of the toxin B subunit. Significantly, these residues are not conserved in STx2, explaining the lack of effectiveness of Mn against STx2. Together our results imply that STx2 uses an evolutionarily distinct trafficking mechanism and that Mn as a potential therapy should be focused on STx/STx1 outbreaks, which account for the vast majority of cases worldwide.

scholarly journals Switching Shiga Toxin (Stx) Type from Stx2d to Stx2a but Not Stx2c Alters Virulence of Stx-Producing Escherichia coli (STEC) Strain B2F1 in Streptomycin (Str)-Treated Mice

Toxins ◽  
2021 ◽  
Vol 13 (1) ◽  
pp. 64
Author(s):  
Beth A. McNichol ◽  
Rebecca A. Bova ◽  
Kieron Torres ◽  
Lan N. Preston ◽  
Angela R. Melton-Celsa
Keyword(s):  
Escherichia Coli ◽  
Amino Acid ◽  
Oral Administration ◽  
Amino Acid Composition ◽  
Shiga Toxin ◽  
Vero Cells ◽  
Toxin B ◽  
B Subunit ◽  
Intestinal Mucus ◽  
Binding Subunit

Shiga toxin (Stx)-producing Escherichia coli (STEC) strain B2F1 produces Stx type 2d, a toxin that becomes more toxic towards Vero cells in the presence of intestinal mucus. STEC that make Stx2d are more pathogenic to streptomycin (Str)-treated mice than most STEC that produce Stx2a or Stx2c. However, purified Stx2d is only 2- or 7-fold more toxic by the intraperitoneal route than Stx2a or Stx2c, respectively. We hypothesized, therefore, that the toxicity differences among Stx2a, Stx2c, and Stx2d occur at the level of delivery from the intestine. To evaluate that hypothesis, we altered the toxin type produced by stx2d+ mouse virulent O91:H21 clinical isolate B2F1 to Stx2a or Stx2c. Because B2F1 encodes two copies of stx2d, we did these studies in a derivative of B2F1 in which stx2d1 was deleted. Although the strains were equivalently virulent to the Str-treated mice at the 1010 dose, the B2F1 strain that produced Stx2a was attenuated relative to the ones that produced Stx2d or Stx2c when administered at 103 CFU/mouse. We next compared the oral toxicities of purified Stx2a, Stx2c, and Stx2d. We found that purified Stx2d is more toxic than Stx2a or Stx2c upon oral administration at 4 µg/mouse. Taken together, these studies suggest that Stx2 toxins are most potent when delivered directly from the bacterium. Furthermore, because Stx2d and Stx2c have the identical amino acid composition in the toxin B subunit, our results indicate that the virulence difference between Stx2a and Stx2d and Stx2c resides in the B or binding subunit of the toxins.

A System for Production and Rapid Purification of Large Amounts of the Shiga Toxin/Shiga-Like Toxin B Subunit

1991 ◽  
pp. 299-301
Author(s):  
Arthur Donohue-Rolfe ◽  
David W. K. Acheson ◽  
Gerald T. Keusch ◽  
Marcia B. Goldberg ◽  
Stephanie A. Boyko ◽  
...  
Keyword(s):  
Shiga Toxin ◽  
Toxin B ◽  
B Subunit ◽  
Rapid Purification

scholarly journals Shiga toxin B-subunit sequential binding to its natural receptor in lipid membranes

2007 ◽  
Vol 1768 (3) ◽  
pp. 628-636 ◽  
Author(s):  
David G. Pina ◽  
Ludger Johannes ◽  
Miguel A.R.B. Castanho
Keyword(s):  
Shiga Toxin ◽  
Lipid Membranes ◽  
Toxin B ◽  
B Subunit ◽  
Sequential Binding

scholarly journals Pathogenesis of shigella diarrhea. XI. Isolation of a shigella toxin-binding glycolipid from rabbit jejunum and HeLa cells and its identification as globotriaosylceramide.

1986 ◽  
Vol 163 (6) ◽  
pp. 1391-1404 ◽  
Author(s):  
M Jacewicz ◽  
H Clausen ◽  
E Nudelman ◽  
A Donohue-Rolfe ◽  
G T Keusch
Keyword(s):  
Hela Cells ◽  
Neutralizing Antibodies ◽  
Specific Binding ◽  
Jejunal Mucosa ◽  
Toxin B ◽  
Toxin Binding ◽  
B Subunit ◽  
Specific Binding Sites ◽  
Rabbit Jejunum ◽  
Group B

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.

Two-dimensional structures of the Shiga toxin B-subunit and of a chimera bound to the glycolipid receptor Gb3

2002 ◽  
Vol 139 (2) ◽  
pp. 113-121 ◽  
Author(s):  
Xavier Hagnerelle ◽  
Célia Plisson ◽  
Olivier Lambert ◽  
Sergio Marco ◽  
Jean Louis Rigaud ◽  
...  
Keyword(s):  
Shiga Toxin ◽  
Two Dimensional ◽  
Toxin B ◽  
B Subunit ◽  
Glycolipid Receptor

scholarly journals Recombinant Shiga Toxin B-Subunit-Keyhole Limpet Hemocyanin Conjugate Vaccine Protects Mice from Shigatoxemia

2005 ◽  
Vol 73 (10) ◽  
pp. 6523-6529 ◽  
Author(s):  
Paola Marcato ◽  
Thomas P. Griener ◽  
George L. Mulvey ◽  
Glen D. Armstrong
Keyword(s):  
Shiga Toxin ◽  
Lethal Dose ◽  
Keyhole Limpet Hemocyanin ◽  
Vaccine Antigen ◽  
Enzyme Linked Immunosorbent Assay ◽  
Toxin B ◽  
B Subunit ◽  
Adjuvant System ◽  
Shiga Toxin 2 ◽  
Uremic Syndrome

ABSTRACT Enterohemorrhagic Escherichia coli (EHEC) causes hemorrhagic colitis in humans and, in a subgroup of infected subjects, a more serious condition called hemolytic-uremic syndrome (HUS). These conditions arise because EHEC produces two antigenically distinct forms of Shiga toxin (Stx), called Stx1 and Stx2. Despite this, the production of Stx2 by virtually all EHEC serotypes and the documented role this toxin plays in HUS make it an attractive vaccine candidate. Previously, we assessed the potential of a purified recombinant Stx2 B-subunit preparation to prevent Shigatoxemia in rabbits. This study revealed that effective immunization could be achieved only if endotoxin was included with the vaccine antigen. Since the presence of endotoxin would be unacceptable in a human vaccine, the object of the studies described herein was to investigate ways to safely augment, in mice, the immunogenicity of the recombinant Stx2 B subunit containing <1 endotoxin unit per ml. The study revealed that sera from mice immunized with such a preparation, conjugated to keyhole limpet hemocyanin and administered with the Ribi adjuvant system, displayed the highest Shiga toxin 2 B-subunit-specific immunoglobulin G1 (IgG1) and IgG2a enzyme-linked immunosorbent assay titers and cytotoxicity-neutralizing activities in Ramos B cells. As well, 100% of the mice vaccinated with this preparation were subsequently protected from a lethal dose of Stx2 holotoxin. These results support further evaluation of a Stx2 B-subunit-based human EHEC vaccine.

Towards a vaccine candidate againstShigella dysenteriae1: expression of the Shiga toxin B-subunit in an attenuatedShigella flexneri aroDcarrier strain

1996 ◽  
Vol 21 (4) ◽  
pp. 277-288 ◽  
Author(s):  
Barbara D. Tzschaschel ◽  
Silke R. Klee ◽  
Victor de Lorenzo ◽  
Kenneth N. Timmis ◽  
Carlos A. Guzmán
Keyword(s):  
Shiga Toxin ◽  
Vaccine Candidate ◽  
Toxin B ◽  
B Subunit
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