Unravelling the rules of outer membrane protein folding: Opportunities for the design of new antibacterial strategies

揭示外膜蛋白折叠规则：设计新抗菌策略的机会

• 批准号: 2602428
• 金额: --
• 依托单位: University of Leeds
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2021
• 资助国家: 英国
• 起止时间: 2021 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2602428
• 关键词: Unravelling; rules; outer; membrane; protein

The folding and assembly of outer membrane proteins (OMPs) is essential to build the cell wall of gram-negative bacteria. How the sequences of OMPs enable their folding into the bacterial OM remains unknown, nor is it understood how the essential B-barrel assembly machinery (BAM), recognises folding OMPs and inserts them into the outer membrane (OM). Understanding OMP folding is not only of immense fundamental interest, but is translationally important as targeting this pathway could lead to new strategies to combat antibiotic resistance in gram-negative pathogens. Building on a recent collaboration with David Baker (Seattle) (Science 2021), we have successfully designed the first synthetic trans membrane beta barrel protein, providing a unique opportunity to compare the folding mechanisms of de novo designed proteins with their naturally evolved counterparts in vitro, and assisted/unassisted by BAM.This project will determine:(i) how the sequence of OMPs enables efficient folding to their highly-stable beta-barrel structures in vitro; (ii) which sequence/structural motifs of OMPs are recognised by BAM;(iii) how the cellular machinery recognizes OMPS and targets them to BAM for folding into the OM. The results will be new fundamental knowledge of how OMPs fold, new insights into how BAM recognises OMPs and potentially, new routs of combat antibiotic resistance by targeting OMP folding.

外膜蛋白（OMPs）的折叠和组装对于革兰氏阴性菌细胞壁的构建至关重要。OMP的序列如何使它们能够折叠成细菌OM仍然是未知的，也不知道基本的B桶组装机器（BAM）如何识别折叠的OMP并将它们插入外膜（OM）。了解OMP折叠不仅具有巨大的根本利益，而且非常重要，因为靶向该途径可能导致对抗革兰氏阴性病原体抗生素耐药性的新策略。基于最近与大卫贝克（西雅图）的合作（Science 2021），我们成功设计了第一个合成的跨膜β桶蛋白，提供了一个独特的机会来比较从头设计的蛋白质与它们在体外自然进化的对应物的折叠机制，并在BAM的辅助/非辅助下。该项目将确定：（i）OMPs序列如何在体外有效折叠成高度稳定的β桶结构;（ii）BAM识别OMPs的哪些序列/结构基序;（iii）细胞机器如何识别OMPs并将其靶向BAM以折叠成OM。这些结果将是关于OMP如何折叠的新的基础知识，关于BAM如何识别OMP的新见解，以及通过靶向OMP折叠来对抗抗生素耐药性的新途径。