Globotriaosylceramide (Gb3) content in HeLa cells is correlated to Shiga toxin-induced cytotoxicity and Gb3 synthase expression

Shin, In-Sun; Ishii, Satoshi; Shin, Jong-Seo; Sung, Kyong-Il; Park, Byung-Sung; Jang, Hyun-Yong; Kim, Byong-Wan

doi:10.5483/bmbrep.2009.42.5.310

Cited by 24 publications

(18 citation statements)

References 27 publications

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“…STx1, secreted by EHEC, has a B subunit identical to STx secreted by Shigella, and the A subunit differs in only one position (17). As expected (18), the lethal dose (LD 50 ) of STx1 was 0.05 ng/ml (Fig. 4A).…”

supporting

confidence: 88%

Manganese Blocks Intracellular Trafficking of Shiga Toxin and Protects Against Shiga Toxicosis

Mukhopadhyay

Linstedt

2012

Science

114

180

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Infections with Shiga toxin (STx)–producing bacteria cause more than a million deaths each year and have no definitive treatment. To exert its cytotoxic effect, STx invades cells through retrograde membrane trafficking, escaping the lysosomal degradative pathway. We found that the widely available metal manganese (Mn2+) blocked endosome-to-Golgi trafficking of STx and caused its degradation in lysosomes. Mn2+ targeted the cycling Golgi protein GPP130, which STx bound in control cells during sorting into Golgi-directed endosomal tubules that bypass lysosomes. In tissue culture cells, treatment with Mn2+ yielded a protection factor of 3800 against STx-induced cell death. Furthermore, mice injected with nontoxic doses of Mn2+ were completely resistant to a lethal STx challenge. Thus, Mn2+ may represent a low-cost therapeutic agent for the treatment of STx infections.

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supporting

confidence: 88%

Manganese Blocks Intracellular Trafficking of Shiga Toxin and Protects Against Shiga Toxicosis

Mukhopadhyay

Linstedt

2012

Science

114

180

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“…These profiles are most likely quite different for the plasma membranes of PBMC versus TZM-bl, and for the envelopes of pseudoviruses (293T endothelial cell-derived), as compared with primary isolates (PBMC-derived). Like PBMC, HeLa-derived cells also express both Gb3 (Shin et al, 2009) and GM3 (Markwell et al, 1984) although degree of expression has been shown to vary with degree of differentiation, passage number, and culture conditions (Markwell et al, 1984). It is known that mitogen activation increases cell surface expression of GSLs (Fantini et al, 1998a; Lund et al, 2006).…”

Section: Discussionmentioning

confidence: 99%

Multivalent dendrimeric compounds containing carbohydrates expressed on immune cells inhibit infection by primary isolates of HIV-1

et al. 2010

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Specific glycosphingolipids (GSL), found on the surface of target immune cells, are recognized as alternate cell surface receptors by the human immunodeficiency virus type 1 (HIV-1) external envelope glycoprotein. In this study, the globotriose and 3’-sialyllactose carbohydrate head groups found on two GSL were covalently attached to a dendrimer core to produce two types of unique multivalent carbohydrates (MVC). These MVC inhibited HIV-1 infection of T cell lines and primary peripheral blood mononuclear cells (PBMC) by T cell line-adapted viruses or primary isolates, with IC50s ranging from 0.1 – 7.4 µg/ml. Inhibition of Env-mediated membrane fusion by MVC was also observed using a dye-transfer assay. These carbohydrate compounds warrant further investigation as a potential new class of HIV-1 entry inhibitors. The data presented also shed light on the role of carbohydrate moieties in HIV-1 virus-host cell interactions.

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“…Stx is an AB 5 toxin consisting of a single A subunit associated with a pentamer of identical B subunits. This pentamer binds to the glycosphingolipid globotriaosylceramide (Gb3) in host cell membranes [4], [5], [6], [7] and delivers the A subunit into the cytoplasm. In the cytoplasm, the enzymatically active A subunit inhibits protein synthesis by cleaving an adenine nucleotide from 28S RNA within the 60S ribosomal subunit, preventing tRNA binding and protein synthesis [8], [9].…”

Section: Introductionmentioning

confidence: 99%

Shiga Toxin Binding to Glycolipids and Glycans

et al. 2012

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BackgroundImmunologically distinct forms of Shiga toxin (Stx1 and Stx2) display different potencies and disease outcomes, likely due to differences in host cell binding. The glycolipid globotriaosylceramide (Gb3) has been reported to be the receptor for both toxins. While there is considerable data to suggest that Gb3 can bind Stx1, binding of Stx2 to Gb3 is variable.MethodologyWe used isothermal titration calorimetry (ITC) and enzyme-linked immunosorbent assay (ELISA) to examine binding of Stx1 and Stx2 to various glycans, glycosphingolipids, and glycosphingolipid mixtures in the presence or absence of membrane components, phosphatidylcholine, and cholesterol. We have also assessed the ability of glycolipids mixtures to neutralize Stx-mediated inhibition of protein synthesis in Vero kidney cells.ResultsBy ITC, Stx1 bound both Pk (the trisaccharide on Gb3) and P (the tetrasaccharide on globotetraosylceramide, Gb4), while Stx2 did not bind to either glycan. Binding to neutral glycolipids individually and in combination was assessed by ELISA. Stx1 bound to glycolipids Gb3 and Gb4, and Gb3 mixed with other neural glycolipids, while Stx2 only bound to Gb3 mixtures. In the presence of phosphatidylcholine and cholesterol, both Stx1 and Stx2 bound well to Gb3 or Gb4 alone or mixed with other neutral glycolipids. Pre-incubation with Gb3 in the presence of phosphatidylcholine and cholesterol neutralized Stx1, but not Stx2 toxicity to Vero cells.ConclusionsStx1 binds primarily to the glycan, but Stx2 binding is influenced by residues in the ceramide portion of Gb3 and the lipid environment. Nanomolar affinities were obtained for both toxins to immobilized glycolipids mixtures, while the effective dose for 50% inhibition (ED50) of protein synthesis was about 10−11 M. The failure of preincubation with Gb3 to protect cells from Stx2 suggests that in addition to glycolipid expression, other cellular components contribute to toxin potency.

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Globotriaosylceramide (Gb3) content in HeLa cells is correlated to Shiga toxin-induced cytotoxicity and Gb3 synthase expression

Cited by 24 publications

References 27 publications

Manganese Blocks Intracellular Trafficking of Shiga Toxin and Protects Against Shiga Toxicosis

Manganese Blocks Intracellular Trafficking of Shiga Toxin and Protects Against Shiga Toxicosis

Multivalent dendrimeric compounds containing carbohydrates expressed on immune cells inhibit infection by primary isolates of HIV-1

Shiga Toxin Binding to Glycolipids and Glycans

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