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Maintaining cell wall integrity in Gram-Negative Bacteria

维持革兰氏阴性细菌细胞壁的完整性

基本信息

批准号：
NE/T014717/1
负责人：
David Ian Roper
金额：
$ 1.48万
依托单位：
University of Warwick
依托单位国家：
英国
项目类别：
Research Grant
财政年份：
2020
资助国家：
英国
起止时间：
2020 至无数据
项目状态：
已结题

来源：
https://gtr.ukri.org/projects?ref=NE%2FT014717%2F1
关键词：
Maintaining cell wall integrity Gram

项目摘要

BBSRC: Christopher Graham: BB/M01116X/1The development of resistance to antibiotics by pathogenic bacteria already undermines modern and future healthcare and has been acknowledged as a worldwide issue with an impact as important as climate change. We are in initial stages of a post-antibiotics era, akin to the pre-19th century world in terms of the danger from infections. Or particular concern in this respect are Gram-negative bacteria since their outer membrane presents a particular barrier to entry of existing and future antibiotics. The pharmaceutical industry has reduced in its capacity and commitment to funding antibiotic discovery in recent years due to complex economic issues, however, a large proportion of successful antibiotics have been those based on targeting the cell walls of bacteria generally. Both Gram-negative bacteria and Gram-positive bacteria have an inner membrane and cell wall peptidoglycan polymer however a third outer layer known as the outer membrane is unique to Gram-negatives. This outer membrane in Gram-negative bacteria's greatest defence against most antibiotics since it provides a significant barrier to their entry as well as having an important biological role in Gram-negative bacterial physiology. Our hypothesis is that the process of cell wall peptidoglycan and outer membrane biogenesis must be linked and if we understood this process then it may lead to the use of combinatorial antibiotics strategies that affect both systems synergistically. This would allow us to truly break down Gram-negative bacterial pathogens, disrupting the outer-membrane, therefore, allowing peptidoglycan affecting drugs to pass. We have preliminary data that shows disruption of the peptidoglycan synthesis system actually causes outer membrane destabilisation and increased bacterial susceptibility, and hypothesise the outer membrane and peptidoglycan layers are created simultaneously and locally through complex interactions.We have established a collaboration with Professor Lori Burrows at McMaster University in Canada who has the specialist genetic tools and facilities to enable us to explore this hypothesis in P. aeuginosa, one of the most important Gram-Negative pathogens. The University of Warwick does not currently have the facilities or expertise to do this but our collaboration with Canada will allow this knowledge to be gained and facilitate a parallel study in E.coli in the future. We aim in this study, to show the co-ordination of the proteins responsible for outer membrane and peptidoglycan synthesis by making bacterial strains in which individual pairs of proteins are specially fluorescently tagged allowing us to use appropriate microscopy to visualise their co-localisation. This will establish at a molecular level how these processes are linked and what proteins may be involved. Previously both outer membrane peptidoglycan biogenesis has been studied as separate events but knowledge gained in this collaboration could be of fundamental biological importance as well as the basis upon which future antimicrobial strategies that target the two processes in parallel are based. The study in Canada will be undertaken by a highly talented UK PhD student who will learn new skills, be exposed to an entirely different research environment and thus gain enormously from this international training opportunity.

BBSRC：克里斯托弗·格雷厄姆：BB/M01116 X/1病原菌对抗生素的耐药性已经破坏了现代和未来的医疗保健，并被认为是一个全球性问题，其影响与气候变化一样重要。我们正处于后抗生素时代的初始阶段，就感染的危险而言，类似于19世纪世纪前的世界。或者在这方面特别关注的是革兰氏阴性细菌，因为它们的外膜对现有和未来的抗生素的进入呈现出特定的屏障。由于复杂的经济问题，近年来制药行业对抗生素发现的资助能力和承诺有所减少，然而，很大一部分成功的抗生素都是基于针对细菌细胞壁的抗生素。革兰氏阴性菌和革兰氏阳性菌都具有内膜和细胞壁肽聚糖聚合物，然而，被称为外膜的第三外层是革兰氏阴性菌所特有的。这种外膜是革兰氏阴性细菌对大多数抗生素的最大防御，因为它为抗生素的进入提供了重要的屏障，并且在革兰氏阴性细菌生理学中具有重要的生物学作用。我们的假设是，细胞壁肽聚糖和外膜生物合成的过程必须联系在一起，如果我们理解了这个过程，那么它可能会导致使用组合抗生素策略，协同影响两个系统。这将使我们能够真正分解革兰氏阴性细菌病原体，破坏外膜，因此，允许肽聚糖影响药物通过。我们有初步的数据表明，肽聚糖合成系统的破坏实际上会导致外膜不稳定和细菌易感性增加，我们与加拿大麦克马斯特大学的洛里伯罗斯教授建立了合作关系，他拥有专业的遗传工具和设施，使我们能够探索这一点铜绿假单胞菌是最重要的革兰氏阴性病原体之一。沃里克大学目前没有这样做的设施或专业知识，但我们与加拿大的合作将使这种知识得以获得，并促进未来在大肠杆菌中的平行研究。我们在这项研究中的目标是，通过制造细菌菌株来显示负责外膜和肽聚糖合成的蛋白质的协调，其中单独的蛋白质对被特别荧光标记，使我们能够使用适当的显微镜来可视化它们的共定位。这将在分子水平上确定这些过程是如何联系在一起的，以及可能涉及哪些蛋白质。以前两个外膜肽聚糖生物合成已被研究作为单独的事件，但在这种合作中获得的知识可能是基本的生物学重要性，以及未来的抗菌策略，目标是两个过程的基础上平行。在加拿大的研究将由一个非常有才华的英国博士生谁将学习新的技能，接触到一个完全不同的研究环境，从而从这个国际培训机会获得巨大的承担。