Elucidating the structures of gliding motility in Bdellovibrio bacteriovorus

蛭弧菌是其他革兰氏阴性细菌的天然捕食者，包括重要的植物和动物病原体，但不能伤害高等生物。这使得蛭弧菌作为一种活的抗菌剂具有巨大的潜力，可应用于医学，动物健康和作物保护。蛭弧菌的特殊生态位意味着它的许多蛋白质广泛适应捕食作用，因此与其他细菌无法比拟。因此，了解这种微观捕食者的生活的适当方法是使用结构生物学技术直接研究其分子机制。本计画将利用生物化学、结构生物学及遗传学技术，探讨滑行运动的机制。滑翔对于蛭弧菌寻找和杀死猎物的能力至关重要，深入了解驱动这种运动的马达将有助于更好地了解其生命周期。我们将开始从结构上研究蛭弧菌的多个滑翔机组成部分，包括单独的滑翔机和相互复合的滑翔机，目的是描述一个完整的滑翔机复合体。这个主要的工作大纲可以分为更小的子项目，由感兴趣的特定组件或滑动复合物（由不同的操纵子编码）定义。另一个目标将是使用细菌双杂交系统和下拉测定来鉴定蛋白质-蛋白质相互作用，以及推定的结合基序的靶向突变，以揭示运动元件拼凑在一起的方式。

Bdellovibrio is a natural predator of other gram-negative bacteria, including important plant and animal pathogens, yet is incapable of harming higher organisms. This gives Bdellovibrio enormous potential as a living antimicrobial with applications in medicine, animal health and crop protection. The extraordinary niche of Bdellovibrio means that many of its proteins are extensively adapted to roles in predation and are therefore incomparable to other bacteria. Thus, an appropriate method of understanding the life of this microscopic predator is by directly investigating its molecular mechanisms using structural biology techniques. This project will utilise biochemistry, structural biology, and genetic techniques to investigate the mechanisms underpinning gliding motility. Gliding is critical to Bdellovibrio's ability to both search for and kill prey, and insight into the motors driving this motility would be instrumental to better understanding its life cycle. We will begin to structurally characterise the components that comprise the multiple gliding machineries belonging to Bdellovibrio, alone and then in complex with one another, with the ambition to characterize a full gliding motor complex. This major work outline can be divided into smaller sub-projects, defined by either the specific component or gliding complex (encoded by distinct operons) of interest. An additional goal will be to use the bacterial two-hybrid system and pull-down assays to identify protein-protein interactions, and targeted mutations of putative binding motifs to reveal the way that the motor elements piece together.