Bacillus Subtilis Chaperone/protease Mechanisms In Metabolic Shutdown

枯草芽孢杆菌伴侣/蛋白酶代谢关闭机制

• 批准号: BB/X001415/1
• 负责人: Rivka Isaacson
• 金额: $ 64.92万
• 依托单位: King's College London
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2023
• 资助国家: 英国
• 起止时间: 2023 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FX001415%2F1
• 关键词: Bacillus; Subtilis; Chaperone; protease; Mechanisms

In many types of bacteria there is a rotary-shredder machine called ClpCP that acts as waste disposal for proteins that are no longer needed. It is built of two parts: ClpC, which is a 6-membered ring that uses energy from ATP hydrolysis to unwind the three dimensional shapes of proteins into a single chain that looks like beads on a wire. This is then fed into ClpP, which takes the form of a stacked pair of 7-membered rings, to be chopped up into small reusable parts. The ClpCP machine is recruited to particular jobs within cells by adaptor proteins. One of these jobs is to destroy many proteins that are important for metabolism, when bacteria form long-lived hardy spores to survive harsh conditions as the spores need to become metabolically inactive. In this study my group will examine a newly discovered protein, MicA, which acts an adaptor to ClpCP during bacterial spore formation. Sporulation is partially responsible for the persistence of 'hospital superbugs' as spores are a long-lived bacterial form, resistant to cleaning agents and thriving in patients depleted of natural gut microflora.We intend to uncover the detailed molecular shapes of MicA when it is bound to ClpCP and substrates using indirect techniques as they are too small to see even using powerful microscopes. We specialise in measuring protein shapes and the way they fit together by producing them artificially in large quantities, with the help of bacteria which act as our 'protein factories'. We then deduce the proteins' molecular structures by processing their behaviour when we bounce X-rays or electrons off them or put them in strong magnetic fields. Each of these techniques has its strengths and weaknesses but our combined approach can yield complementary information filling in the gaps left by using just one of the methods. Collaborating with microbiologists, electron microscopists and chaperone scientists, we will feed information into each others' experiments to build up a mechanistic picture of this important step in bacterial spore formation. For example, if we identify a mutation in one of our proteins that makes it bind more tightly to its partner our collaborator can make the same mutation within bacteria to test whether it has the predicted effect in living systems.By solving this jigsaw puzzle we hope to be in a stronger position to design novel antibiotics to attack the increasing problem of bacterial drug resistance and the project also has longer term implications for understanding metabolism and protein quality control in many aspects of health and disease.

在许多类型的细菌中，有一台称为CLPCP的旋转式机器，可作为不再需要的蛋白质的废物处理。它由两个部分构建：CLPC，它是一个6元的环，它利用ATP水解的能量将蛋白质的三维形状放在单个链中，将蛋白质的三维形状放在一条看起来像电线上的珠子的单个链中。然后将其馈入CLPP，以一对堆叠的7元环的形式将其切成可重复使用的小部分。 CLPCP机器通过衔接蛋白招募到细胞内的特定作业。这些工作之一是消灭许多对新陈代谢重要的蛋白质，当细菌形成长寿的耐硬孢子以生存恶劣的条件，因为孢子需要代谢不活跃。在这项研究中，我的小组将检查新发现的蛋白质云母，该蛋白质在细菌孢子形成过程中起着对CLPCP的适配器。孢子形成部分负责“医院超级细菌”的持续性，因为孢子是一种长寿形式，对清洁剂的耐药性和在天然肠道微生物中耗尽的患者中蓬勃发展。我们打算在使用Indiques clpcp and-crictiques by-simby的情况下，发现米云的详细分子形状，即它们既有小小的技术，又是小小的技术。我们专注于测量蛋白质形状以及它们通过大量人工产生的蛋白质形状，借助细菌作为我们的“蛋白质工厂”。然后，当我们弹跳X射线或电子或将它们放置在强磁场中时，我们通过处理其行为来推断蛋白质的分子结构。这些技术中的每一个都有其优点和劣势，但是我们的合并方法可以通过仅使用其中一种方法来产生互补的信息填充所留下的空白。与微生物学家，电子显微镜医生和伴侣科学家合作，我们将向彼此的实验提供信息，以建立细菌孢子形成中这一重要步骤的机械图。例如，如果我们在其中一种蛋白质中确定突变，使其与伴侣更紧密地结合，我们的合作者可以在细菌中制造相同的突变，以测试它是否在生活系统中具有预测的效果。通过解决这种拼图难题，我们希望我们希望在设计中具有更强的抗生素来攻击较长的抗药性问题，以攻击较长的疗法，并在较长的范围内构成了较长的材料，并且在较长的范围内构成了良好的质量，该材料的质量是细菌的良好问题。和疾病。