Oligomerization of Clostridium perfringens epsilon toxin is dependent upon caveolins 1 and 2.
Oligomerization of Clostridium perfringens epsilon toxin is dependent upon caveolins 1 and 2.
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DOI：
10.1371/journal.pone.0046866
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
McClain MS
中科院分区：
文献类型：
--
作者：
Fennessey CM;Sheng J;Rubin DH;McClain MS
来自多项研究的证据表明，产气荚膜梭菌ε-毒素是一种成孔毒素，在敏感细胞的质膜中组装成寡聚复合物。在以前的研究中，我们使用基因陷阱诱变，以确定哺乳动物的毒素活性，包括小窝蛋白-2（CAV 2）的因素。在这项研究中，我们证明了小窝蛋白-2及其相互作用伙伴，小窝蛋白-1（CAV 1），在ε-毒素诱导的细胞毒性的重要性。在毒素敏感的人肾细胞系ACHN中使用CAV 2特异性shRNA，我们证实缺乏CAV 2的细胞对ε-毒素的抵抗力增强。同样，使用CAV 1特异性shRNA，我们证明了CAV 1缺陷的细胞也表现出对毒素的抵抗力增加。来自ε-毒素处理的ACHN细胞的CAV 1和CAV 2的免疫沉淀证明CAV 1和CAV 2与毒素相互作用。此外，蓝色-非变性PAGE表明毒素和小窝蛋白是670 kDa蛋白复合物的组分。虽然ε-毒素结合在小窝蛋白缺陷细胞中仅轻微扰动，但毒素的寡聚化在CAV 1和CAV 2缺陷细胞中均显著减少。这些结果表明，CAV 1和CAV-2通过促进毒素寡聚化来增强ε-毒素诱导的细胞毒性-这是孔形成和细胞死亡所必需的事件。
Evidence from multiple studies suggests that Clostridium perfringens ε-toxin is a pore-forming toxin, assembling into oligomeric complexes in the plasma membrane of sensitive cells. In a previous study, we used gene-trap mutagenesis to identify mammalian factors contributing to toxin activity, including caveolin-2 (CAV2). In this study, we demonstrate the importance of caveolin-2 and its interaction partner, caveolin-1 (CAV1), in ε-toxin-induced cytotoxicity. Using CAV2-specific shRNA in a toxin-sensitive human kidney cell line, ACHN, we confirmed that cells deficient in CAV2 exhibit increased resistance to ε-toxin. Similarly, using CAV1-specific shRNA, we demonstrate that cells deficient in CAV1 also exhibit increased resistance to the toxin. Immunoprecipitation of CAV1 and CAV2 from ε-toxin-treated ACHN cells demonstrated interaction of both CAV1 and -2 with the toxin. Furthermore, blue-native PAGE indicated that the toxin and caveolins were components of a 670 kDa protein complex. Although ε-toxin binding was only slightly perturbed in caveolin-deficient cells, oligomerization of the toxin was dramatically reduced in both CAV1- and CAV2-deficient cells. These results indicate that CAV1 and -2 potentiate ε-toxin induced cytotoxicity by promoting toxin oligomerization – an event which is requisite for pore formation and, by extension, cell death.
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影响因子：
4
作者：
Hoffmann, Christine;Berking, Anne;Hauck, Christof R.
通讯作者：
Hauck, Christof R.
影响因子：
64.5
作者：
Gurcel, Laure;Abrami, Laurence;van der Goot, F. Gisou
通讯作者：
van der Goot, F. Gisou
影响因子：
8
作者：
Galdiero, S;Gouaux, E
通讯作者：
Gouaux, E
质膜微域充当浓度平台，可促进气氛蛋白中毒。
DOI：
10.1083/jcb.147.1.175
发表时间：
1999-10-04
期刊：
The Journal of cell biology
影响因子：
--
作者：
Abrami L;van Der Goot FG
通讯作者：
van Der Goot FG
影响因子：
4.8
作者：
Garcia-Saez, Ana J.;Buschhorn, Sabine B.;Schwille, Petra
通讯作者：
Schwille, Petra