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首页> 外文期刊>Journal of Molecular Biology >Ion Conductance of the Stem of the Anthrax Toxin Channel during Lethal Factor Translocation
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Ion Conductance of the Stem of the Anthrax Toxin Channel during Lethal Factor Translocation

机译：致死因子易位过程中炭疽毒素通道茎的离子电导

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摘要

The tripartite anthrax toxin consists of protective antigen, lethal factor (LF), and edema factor. PA(63) (the 63-kDa, C-terminal part of protective antigen) forms heptameric channels in cell membranes that allow for the transport of LF and edema factor into the cytosol. These channels are mushroom shaped, with a ring of seven phenylalanine residues (known as the phenylalanine clamp) lining the junction between the cap and the stem. It is known that when LF is translocated through the channel, the phenylalanine clamp creates a seal that causes an essentially complete block of conduction. In order to examine ion conductance in the stem of the channel, we used Venus yellow fluorescent protein as a molecular stopper to trap LFN (the 30-kDa, 263-residue N-terminal segment of LF), as well as various truncated constructs of LFN, in mutant channels in which the phenylalanine clamp residues were mutated to alanines. Here we present evidence that ion movement occurs within the channel stem (but is stopped, of course, at the phenylalanine clamp) during protein translocation. Furthermore, we also propose that the lower region of the stem plays an important role in securing peptide chains during translocation. (C) 2014 Elsevier Ltd. All rights reserved.
机译：三方炭疽毒素由保护性抗原，致死因子（LF）和浮肿因子组成。 PA（63）（保护性抗原的63 kDa C端部分）在细胞膜上形成七聚体通道，从而可以将LF和水肿因子转运到细胞质中。这些通道是蘑菇形的，在帽和茎之间的连接处有一个七个苯丙氨酸残基的环（称为苯丙氨酸钳）。众所周知，当LF通过通道移位时，苯丙氨酸钳会形成一个密封，从而导致基本上完全的传导阻滞。为了检查通道干中的离子电导，我们使用金星黄色荧光蛋白作为分子塞子来捕获LFN（LF的30kDa，263个残基的N末端片段），以及各种截短的构建体。 LFN，在苯丙氨酸钳位残基突变为丙氨酸的突变通道中。在这里，我们提供证据表明，在蛋白质移位过程中，离子运动发生在通道杆内（但当然会在苯丙氨酸钳位处停止）。此外，我们还提出茎的下部区域在转运期间在确保肽链中起重要作用。 （C）2014 Elsevier Ltd.保留所有权利。

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