Bacterial modification of carbohydrates; structural and functional analysis of a key virulence strategy.

许多细菌病原体都有一个增强毒力和持久性的共同机制;它们产生修饰细菌细胞表面碳水化合物的酶。所述修饰可以帮助逃避免疫系统（例如沙门氏菌属中的O-抗原的修饰），增强生物膜形成（金黄色葡萄球菌）或提供保护免受渗透压（大肠杆菌）;抑制这些酶可能导致新的抗生素。为了开发这些蛋白质作为潜在的新的治疗靶点，有许多问题需要回答的结构和功能之间的关系。本申请的目的是研究该蛋白质家族的成员OpgC，其在革兰氏阴性细菌病原体中使周质葡聚糖琥珀酰化。该项目将采用结构生物学、生物物理学和分子微生物学的跨学科方法和尖端技术。与目前许多药物靶点一样，OpgC是一种完整的膜蛋白，因此该项目也将为这些具有挑战性的蛋白质提供宝贵的技术经验。您将接受重组蛋白的培训，包括膜蛋白，表达和纯化，使用新型酰基供体模拟物和生物物理/结构表征开发膜蛋白的功能测定，包括NMR光谱学和X射线晶体学。生物学系的技术设施还拥有广泛的设施，包括用于表征改性底物的最先进的质谱仪。实验室会议，研讨会系列，minisymopsia和参与和发展推广活动的机会将帮助您开发广泛的科学和可转移的技能。有一个充满活力的博士社区，举办咖啡和职业等活动。您将共同监督，并将在专门从事生物化学，结构生物学和微生物学的实验室进行工作。该项目适合于对生物化学有很强背景和兴趣的申请人，以及对细菌致病机制的分子基础有整体兴趣的申请人。

Many bacterial pathogens share a common mechanism for enhancing virulence and persistence; they produce enzymes that modify carbohydrates on the bacterial cell surface. The modification may aid evasion of the immune system (for example modification of the O-antigen in Salmonella sp.), enhance biofilm formation (Staphylococcus aureus) or provide protection from osmotic stress (Escherichia coli); inhibiting these enzymes could lead to new antibiotics. With a view to developing these proteins as potential new therapeutic targets, there are many questions that need to be answered regarding the relationship between structure and function. The aim of this application is to investigate a member of this protein family, OpgC, that succinylates periplasmic glucans in Gram-negative bacterial pathogens. The project will employ an interdisciplinary approach and cutting edge techniques in structural biology, biophysics and molecular microbiology. Like many current drug targets, OpgC is an integral membrane protein thus this project will also provide valuable experience in techniques for working with these challenging proteins. You will receive training in recombinant protein, including membrane protein, expression and purification, the development of functional assays for membrane proteins using novel acyl donor mimics and biophysical/structural characterisation including, as appropriate, NMR spectroscopy and X-ray crystallography. The Technology Facility in the Department of Biology also houses a wide range of facilities including state-of- the-art mass spectrometry for characterising modified substrates. Lab meetings, seminar series, minisymposia and the opportunity to participate in and develop outreach activities will help you develop a broad range of scientific and transferable skills. There is a vibrant PhD community that hosts events like Coffee and Careers. You will be co-supervised and will carry the work out in labs specializing in biochemistry, structural biology and microbiology. This project is suitable for applicants with a strong background and interest in biochemistry and an overall interest in the molecular basis of bacterial pathogenesis.