Designed coiled-coil peptides inhibit the type three secretion system of enteropathogenic Escherichia coli.
Designed coiled-coil peptides inhibit the type three secretion system of enteropathogenic Escherichia coli.
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DOI：
10.1371/journal.pone.0009046
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发表时间：
2010-02-04
期刊：
影响因子：
3.7
通讯作者：
Navarro-Garcia F
中科院分区：
文献类型：
--
作者：
Larzábal M;Mercado EC;Vilte DA;Salazar-González H;Cataldi A;Navarro-Garcia F
肠致病性大肠埃希菌(EPEC)和肠出血性大肠杆菌(EHEC)是与人类疾病相关的两类大肠杆菌。这两种致病类型的一个主要毒力因素是三型分泌系统(TTSS)的表达，TTSS负责它们与肠粘膜的粘连能力，导致一种特征性的附着和消失病变(A/E)。TTSS将效应蛋白直接转移到宿主细胞中，从而颠覆哺乳动物细胞的生物化学。我们检查了旨在抑制TTSS的合成肽。Coila和CoilB多肽(代表转位蛋白EspA的卷曲区域)和Coild多肽(对应于针状蛋白EscF的卷曲区域)能有效地抑制EPEC E2348/69菌株对TTSS依赖的红细胞的溶血。Coila和CoilB多肽还减少了同一菌株在Hep-2细胞中肌动蛋白底座的形成，并损害了TTSS介导的蛋白质转位到上皮细胞中。有趣的是，Coila和CoilB能够阻止EspA组装，破坏TTSS的稳定性，从而导致TIR易位。Coila或CoilB多肽对EspA聚合的这种阻断也抑制了免疫印迹检测到的EspB和EspD的正确递送。有趣的是，与Coila多肽孵育的细菌的电子显微镜显示EspA细丝长度缩短。我们的数据表明，盘状多肽可以阻止TTSS装置的组装，从而阻止TTSS装置的功能，并表明这些多肽可能提供一个有吸引力的工具来阻断EPEC和EHEC的发病。
Enteropathogenic E. coli (EPEC) and enterohemorrhagic E. coli (EHEC) are two categories of E. coli strains associated with human disease. A major virulence factor of both pathotypes is the expression of a type three secretion system (TTSS), responsible for their ability to adhere to gut mucosa causing a characteristic attaching and effacing lesion (A/E). The TTSS translocates effector proteins directly into the host cell that subvert mammalian cell biochemistry. We examined synthetic peptides designed to inhibit the TTSS. CoilA and CoilB peptides, both representing coiled-coil regions of the translocator protein EspA, and CoilD peptide, corresponding to a coiled–coil region of the needle protein EscF, were effective in inhibiting the TTSS dependent hemolysis of red blood cells by the EPEC E2348/69 strain. CoilA and CoilB peptides also reduced the formation of actin pedestals by the same strain in HEp-2 cells and impaired the TTSS-mediated protein translocation into the epithelial cell. Interestingly, CoilA and CoilB were able to block EspA assembly, destabilizing the TTSS and thereby Tir translocation. This blockage of EspA polymerization by CoilA or CoilB peptides, also inhibited the correct delivery of EspB and EspD as detected by immunoblotting. Interestingly, electron microscopy of bacteria incubated with the CoilA peptide showed a reduction of the length of EspA filaments. Our data indicate that coiled-coil peptides can prevent the assembly and thus the functionality of the TTSS apparatus and suggest that these peptides could provide an attractive tool to block EPEC and EHEC pathogenesis.
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影响因子：
4.8
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