Abstract
Epsilon-toxin (ET) of Clostridiumperfringens, which causes fatal enterotoxemia in ungulates, was previously shown to bind to and form a heptameric pore within the detergent-resistant membranes (DRMs) of MDCK cells. Depletion of cholesterol has also been shown to decrease the cytotoxicity of ET and its heptamerization. In this study, we investigated the effects of changes in sphingolipids, other DRM components of MDCK cells, on the cells' susceptibility to ET. Treatment with fumonisin B1 and PDMP, inhibitors of sphingolipid and glycosphingolipid syntheses, respectively, increased the susceptibility, while D609, a sphingomyelin synthesis inhibitor, had the opposite effect. The exogenous addition of ganglioside GM1 dramatically decreased the ET binding, heptamerization and cytotoxicity. These effects were shown not to be due to ET binding to GM1 or to denaturation of ET. We also found that the ET cytotoxicity towards MDCK cells decreased with an increase in culture time. In accordance with the resistance observed for prolonged cultured cells, GM3, a major ganglioside component, increased and sialidase treatment increased their susceptibility. These results suggest that membrane-anchored sialic acid of GM3 within DRMs inhibits ET binding, leading to prevention of the heptamerization of ET and cell death. It is also suggested that sialidase produced by this organism aids the targeting of ET to MDCK cells.
| Original language | English |
|---|---|
| Pages (from-to) | 245-253 |
| Number of pages | 9 |
| Journal | Microbiology and Immunology |
| Volume | 49 |
| Issue number | 3 |
| Publication status | Published - 2005 |
| Externally published | Yes |
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Keywords
- Clostridium perfringens
- Detergent-resistant membranes
- Epsilon-toxin
- Ganglioside
ASJC Scopus subject areas
- Immunology and Microbiology(all)
- Microbiology (medical)
- Microbiology
Cite this
Changes in ganglioside content affect the binding of Clostridium perfringens epsilon-toxin to detergent-resistant membranes of Madin-Darby canine kidney cells. / Shimamoto, Seiko; Tamai, Eiji; Matsushita, Osamu; Minami, Junzaburo; Okabe, Akinobu; Miyata, Shigeru.
In: Microbiology and Immunology, Vol. 49, No. 3, 2005, p. 245-253.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Changes in ganglioside content affect the binding of Clostridium perfringens epsilon-toxin to detergent-resistant membranes of Madin-Darby canine kidney cells
AU - Shimamoto, Seiko
AU - Tamai, Eiji
AU - Matsushita, Osamu
AU - Minami, Junzaburo
AU - Okabe, Akinobu
AU - Miyata, Shigeru
PY - 2005
Y1 - 2005
N2 - Epsilon-toxin (ET) of Clostridiumperfringens, which causes fatal enterotoxemia in ungulates, was previously shown to bind to and form a heptameric pore within the detergent-resistant membranes (DRMs) of MDCK cells. Depletion of cholesterol has also been shown to decrease the cytotoxicity of ET and its heptamerization. In this study, we investigated the effects of changes in sphingolipids, other DRM components of MDCK cells, on the cells' susceptibility to ET. Treatment with fumonisin B1 and PDMP, inhibitors of sphingolipid and glycosphingolipid syntheses, respectively, increased the susceptibility, while D609, a sphingomyelin synthesis inhibitor, had the opposite effect. The exogenous addition of ganglioside GM1 dramatically decreased the ET binding, heptamerization and cytotoxicity. These effects were shown not to be due to ET binding to GM1 or to denaturation of ET. We also found that the ET cytotoxicity towards MDCK cells decreased with an increase in culture time. In accordance with the resistance observed for prolonged cultured cells, GM3, a major ganglioside component, increased and sialidase treatment increased their susceptibility. These results suggest that membrane-anchored sialic acid of GM3 within DRMs inhibits ET binding, leading to prevention of the heptamerization of ET and cell death. It is also suggested that sialidase produced by this organism aids the targeting of ET to MDCK cells.
AB - Epsilon-toxin (ET) of Clostridiumperfringens, which causes fatal enterotoxemia in ungulates, was previously shown to bind to and form a heptameric pore within the detergent-resistant membranes (DRMs) of MDCK cells. Depletion of cholesterol has also been shown to decrease the cytotoxicity of ET and its heptamerization. In this study, we investigated the effects of changes in sphingolipids, other DRM components of MDCK cells, on the cells' susceptibility to ET. Treatment with fumonisin B1 and PDMP, inhibitors of sphingolipid and glycosphingolipid syntheses, respectively, increased the susceptibility, while D609, a sphingomyelin synthesis inhibitor, had the opposite effect. The exogenous addition of ganglioside GM1 dramatically decreased the ET binding, heptamerization and cytotoxicity. These effects were shown not to be due to ET binding to GM1 or to denaturation of ET. We also found that the ET cytotoxicity towards MDCK cells decreased with an increase in culture time. In accordance with the resistance observed for prolonged cultured cells, GM3, a major ganglioside component, increased and sialidase treatment increased their susceptibility. These results suggest that membrane-anchored sialic acid of GM3 within DRMs inhibits ET binding, leading to prevention of the heptamerization of ET and cell death. It is also suggested that sialidase produced by this organism aids the targeting of ET to MDCK cells.
KW - Clostridium perfringens
KW - Detergent-resistant membranes
KW - Epsilon-toxin
KW - Ganglioside
UR - http://www.scopus.com/inward/record.url?scp=17044432312&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=17044432312&partnerID=8YFLogxK
M3 - Article
C2 - 15781998
AN - SCOPUS:17044432312
VL - 49
SP - 245
EP - 253
JO - Microbiology and Immunology
JF - Microbiology and Immunology
SN - 0385-5600
IS - 3
ER -