Biological activity of a C-terminal fragment of Pasteurella multocida toxin
Biological activity of a C-terminal fragment of Pasteurella multocida toxin
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DOI：
10.1128/iai.69.6.3628-3634.2001
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发表时间：
2001-06-01
影响因子：
3.1
通讯作者：
Aktories, K
中科院分区：
文献类型：
--
作者：
Busch, C;Orth, J;Aktories, K
多杀性巴氏杆菌PMT的蛋白毒素是一种有效的有丝分裂原和磷脂酶C β的激活剂。在这项研究中，不同的毒素片段进行了研究，一个C-末端片段，包括氨基酸581至1285（PMT 581 C）的构建，这是无活性的完整的牛胚肺（EBL）细胞加入培养基后，但引起重组的肌动蛋白细胞骨架和四舍五入的细胞时，通过电穿孔引入细胞。作为全毒素，毒素片段PMT 581 C在通过电穿孔引入细胞后诱导总肌醇磷酸水平的增加。短于PMT 581 C的C-末端片段以及N-末端片段无活性。全毒素中丝氨酸与半胱氨酸-1165的交换导致增加磷酸肌醇水平的能力完全丧失。相应地，突变的毒素片段PMT581C.C1165S在通过电穿孔引入细胞后无活性，表明Cys-1165在毒素的生物活性中起重要作用。
The protein toxin of Pasteurella multocida PMT is a potent mitogen and activator of phospholipase C beta. In this study different toxin fragments were investigated A C-terminal fragment encompassing amino acids 581 through 1285 (PMT581C) was constructed, which was inactive toward intact embryonic bovine lung (EBL) cells after addition to culture medium but caused reorganization of the actin cytoskeleton and rounding up of cells when introduced into the cells by electroporation. As the holotoxin, the toxin fragment PMT581C induced an increase in total inositol phosphate levels after introduction into the cell by electroporation. A C-terminal fragment shorter than PMT581C as well as N-terminal fragments were inactive. Exchange of cysteine-1165 for serine in the holotoxin resulted in a complete loss of the ability to increase inositol phosphate levels. Correspondingly, the mutated toxin fragment PMT581C.C1165S was inactive after cell introduction by electroporation, suggesting an essential role of Cys-1165 in the biological activity of the toxin.