...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Journal of bacteriology >Intracellular Immunization of Prokaryotic Cells against a Bacteriotoxin
【24h】

Intracellular Immunization of Prokaryotic Cells against a Bacteriotoxin

机译：针对细菌毒素的原核细胞的细胞内免疫

获取原文
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Intracellularly expressed antibodies have been designed to bind and inactivate target molecules inside eukaryotic cells. Here we report that an antibody fragment can be used to probe the periplasmic localization of the colicin A N-terminal domain. Colicins form voltage-gated ion channels in the inner membrane of Escherichia coli. To reach their target, they bind to a receptor located on the outer membrane and then are translocated through the envelope. The N-terminal domain of colicins is involved in the translocation step and therefore is thought to interact with proteins of the translocation system. To compete with this system, a single-chain variable fragment (scFv) directed against the N-terminal domain of the colicin A was synthesized and exported into the periplasmic space of E. coli. The periplasmic scFv inhibited the lethal activity of colicin A and had no effect on the lethal activity of other colicins. Moreover, the scFv was able to specifically inactivate hybrid colicins possessing the colicin A N-terminal domain without affecting their receptor binding. Hence, the periplasmic scFv prevents the translocation of colicin A and probably its interaction with import machinery. This indicates that the N-terminal domain of the toxin is accessible in the periplasm. Moreover, we show that production of antibody fragments to interfere with a biological function can be applied to prokaryotic systems.
机译：细胞内表达的抗体已被设计为结合并灭活真核细胞内的靶分子。在这里，我们报告抗体片段可用于探测大肠菌素A N末端域的周质定位。大肠菌素在大肠埃希菌的内膜中形成电压门控离子通道。为了达到目标，它们与位于外膜上的受体结合，然后通过包膜移位。大肠菌素的N末端结构域参与了易位步骤，因此被认为与易位系统的蛋白质相互作用。为了与该系统竞争，合成了针对大肠菌素A N末端结构域的单链可变片段（scFv），并输出到 E的周质空间中。大肠杆菌。周质scFv抑制大肠菌素A的致死活性，而对其他大肠菌素的致死活性没有影响。此外，scFv能够特异性地灭活具有大肠素A N末端结构域的杂交大肠素，而不会影响其受体结合。因此，周质scFv阻止了大肠菌素A的移位，并可能阻止了其与进口机器的相互作用。这表明毒素的N-末端结构域在周质中可及。而且，我们显示出产生干扰生物功能的抗体片段的方法可以应用于原核系统。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号