Pushing the envelope: atomic force microscopy imaging of the bacterial outer membrane during growth and division

挑战极限：生长和分裂过程中细菌外膜的原子力显微镜成像

• 批准号: BB/X00760X/1
• 负责人: Bart Hoogenboom
• 金额: $ 61.35万
• 依托单位: University College London
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2024
• 资助国家: 英国
• 起止时间: 2024 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FX00760X%2F1
• 关键词: Pushing; envelope; atomic; force; microscopy

Gram-negative bacteria are among the microbes with the highest potential to develop resistance against commonly used antibiotics, such that previously harmless infections can become severe and potentially life-threatening diseases. This is in part because these bacteria (including A. baumannii, P. aeruginosa, and enterobacteriaceae such as Salmonella and E. coli) are protected by an outer membrane that limits entry of antibiotics into the cell.Based on light - and particularly fluorescence - microscopy, it has been suggested that this outer membrane organises its building blocks in rather sophisticated ways, including the formation of protein islands that may also play a role in how efficiently bacteria clear antibiotics. Quite generally, light microscopy has been a most powerful tool to discover and understand biological phenomena that define the life, growth, division, and death of cells. Yet its resolution on living cells is mostly insufficient to resolve cells at molecular length scales.Atomic force microscopy (AFM) is an alternative technique that can probe single molecules by gently tracing their contours with a sharp probe. We have recently used AFM to resolve the outer membrane of living bacteria at molecular-scale resolution, thereby revealing how the membrane segregates into different protein-enriched and lipid-enriched domains.Here, noting that bacteria can divide every ~20 minutes under favourable conditions, we propose to develop methods that will enable us to carry out such AFM experiments on growing and dividing cells, with the aim to better understand how the outer membrane facilitates/adapts to bacterial growth and division, including the synthesis and insertion of new membrane components.

革兰氏阴性细菌是具有对常用抗生素抗药性的最高潜力的微生物之一，因此以前无害的感染可能会成为严重且潜在的威胁生命的疾病。这部分是因为这些细菌（包括baumannii，铜绿假单胞菌和肠杆菌科，例如沙门氏菌和大肠杆菌）受到外膜的保护，该外膜限制了抗生素进入细胞的外膜。基于光线的细胞。在细菌清除抗生素方面的作用。通常，光学显微镜是发现和理解定义细胞生命，生长，分裂和死亡的生物学现象的最强大工具。然而，其在活细胞上的分辨率大多不足以在分子长度尺度上解析细胞。原子力显微镜（AFM）是一种替代技术，可以通过用尖锐的探针轻轻跟踪其轮廓来探测单分子。我们最近使用AFM来解决分子尺度分辨率的生物细菌的外膜，从而揭示了膜如何将膜分离为不同的蛋白质增强和脂质增强的结构域。外膜促进/适应细菌生长和分裂，包括合成和插入新的膜成分。