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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Single molecule detection of intermediates during botulinum neurotoxin translocation across membranes
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Single molecule detection of intermediates during botulinum neurotoxin translocation across membranes

机译：肉毒杆菌神经毒素跨膜转运过程中中间体的单分子检测

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摘要

The dynamics of Clostridium botulinum neurotoxins (BoNTs) protein-translocation across membranes was investigated by using a single molecule assay with millisecond resolution on excised patches of neuronal cells. Translocation of BoNT/A light chain (LC) by heavy chain (HC) was observed in real time as an increase of channel conductance: the HC channel is occluded by the LC during transit, then unoccluded after completion of translocation and release of LC-cargo. We identified an entirely unknown succession of intermediate conductance stages during LC translocation. For the single-chain BoNT/E, by contrast to the di-chain BoNT/A, we demonstrate that productive translocation requires proteolysis of the LC cargo from the HC chaperone. We propose a model for the set of protein-protein interactions between translocase and cargo at each step of translocation that supports the notion of an interdependent, tight interplay between the HC chaperone and the LC cargo preventing LC aggregation and dictating the outcome of translocation: productive passage of cargo or abortive channel occlusion by cargo.
机译：肉毒梭菌神经毒素（BoNTs）跨膜蛋白转运的动力学是通过使用毫秒分辨率的单分子测定法对切除的神经元细胞膜片进行研究的。随着通道电导的增加，实时观察到BoNT / A轻链（LC）通过重链（HC）的转运：HC通道在转运过程中被LC阻塞，然后在完成转运和释放LC-之后不阻塞货物。我们确定了LC移位期间中间电导阶段的完全未知的继承。对于单链BoNT / E，与双链BoNT / A相比，我们证明了生产易位需要从HC伴侣得到的LC货物进行蛋白水解。我们提出了在易位的每一步中，转位酶与货物之间的蛋白质-蛋白质相互作用的模型，该模型支持HC分子伴侣和LC货物之间相互依存的紧密相互作用的概念，从而防止LC聚集并决定了易位的结果：货物通过或货物被流产通道阻塞。

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