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首页> 外文期刊>Biochemistry >Formation of a biologically active toxin complex of the binary Clostridium botulinum C2 toxin without cell membrane interaction
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Formation of a biologically active toxin complex of the binary Clostridium botulinum C2 toxin without cell membrane interaction

机译：没有细胞膜相互作用的二元肉毒梭菌C2毒素的生物活性毒素复合物的形成

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摘要

Clostridium botulinum produces a binary toxin, which is composed of two separate proteins. The enzyme component, C2I, is an ADP-ribosyltransferase which modifies G-actin of eukaryotic cells. The proteolytically activated binding/translocation component, C2IIa, forms ring-shaped heptamers, which bind to cell receptors and mediate the transport of C2I into the cytosol of target cells. According to the current model, receptor-bound C2IIa serves as a docking platform for C2I on the cell surface. Following assembly of C2I, the toxin complex is taken up via receptor-mediated endocytosis, and finally, C2IIa facilitates translocation of C2I from acidic endosomes into the cytosol. Our data support an alternative scenario for the early steps of interaction of the C2 toxin and eukaryotic cells, due to the fact that C2IIa and C2I can interact prior to binding of the toxin to the cell surface. The C2IIa-C2I complex, which was formed in a cell-free system, was detected by native gel electrophoresis and subsequent immunoblot analysis or radiolabeling methods. The preformed C2 toxin complex ADP-ribosylated actin in vitro and induced cell rounding. The interaction of C2I with C2IIa did not enhance the binding of C2IIa to the cellular receptor. Intoxication of Vero cells and of human colon carcinoma cells (CaCo-2) was significantly enhanced when the preformed toxin complex was added to cultured cells as compared to addition of the single components.
机译：肉毒梭菌产生一种由两种不同的蛋白质组成的二元毒素。酶成分C2I是ADP-核糖基转移酶，可修饰真核细胞的G-肌动蛋白。蛋白水解激活的结合/转运成分C2IIa形成环形七聚体，该七聚体与细胞受体结合并介导C2I转运到靶细胞的细胞质中。根据当前模型，受体结合的C2IIa充当细胞表面上C2I的对接平台。 C2I组装后，毒素复合物通过受体介导的内吞作用吸收，最后，C2IIa促进C2I从酸性内体向胞质溶胶的转运。由于C2IIa和C2I可以在毒素结合到细胞表面之前相互作用，因此我们的数据为C2毒素与真核细胞相互作用的早期步骤提供了一种替代方案。通过天然凝胶电泳和随后的免疫印迹分析或放射性标记方法检测在无细胞系统中形成的C2IIa-C2I复合物。预先形成的C2毒素复合物ADP-核糖基肌动蛋白在体外并诱导细胞变圆。 C2I与C2IIa的相互作用不会增强C2IIa与细胞受体的结合。当向培养的细胞中添加预先形成的毒素复合物时，与添加单个成分相比，Vero细胞和人结肠癌细胞（CaCo-2）的中毒显着增强。

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