Structure-based drug design against a biosecurity pathogen BB/M009122/1

针对生物安全病原体的基于结构的药物设计 BB/M009122/1

• 批准号: 2579050
• 金额: --
• 依托单位: UNIVERSITY OF EXETER
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2020
• 资助国家: 英国
• 起止时间: 2020 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2579050
• 关键词: Structure; based; drug; design; against

Coxiella burnetii is the causative agent of Q-fever, a zoonotic disease endemic globally and particularly in the South-West in the UK. C. burnetii is an obligate pathogen of ruminants: in livestock it causes abortions and still-births, with obvious economic consequences. C. burnetii is an opportunistic pathogen of humans with a very low infectious dose. Q-fever causes serious flu-like symptoms. Whilst these are usually self-resolving, some patients require hospital treatment, and some of these go on to suffer serious complications (e.g. myocarditis). C. burnetii is one of the causative agents of the so-called "desert fever" suffered by UK service personal in the Middle East, and has been associated with chronic fatigue syndrome. There is a clear need for effective treatments for C. burnetii. We are part of an ongoing (BBSRC funded) programme to produce a vaccine using the polysaccharide O-antigen of C. burnetii, the best current vaccine candidate. This O-antigen contains two highly unusual sugars (virenose and dihydrohydroxystreptose). We have identified several of the biosynthetic proteins that support our hypotheses on the biosynthesis. These enzymes display at least two novel enzyme activities, which may provide excellent opportunities to make specific inhibitors. We have protocols to prepare proteins in high quantities, which should be sufficient to determine the protein structures. The main supervisor will collaborate with Diamond Light Source to use the upcoming micro-electron diffraction method as part of this structure solution.In this project, the student will select the most scientifically interesting proteins from the biosynthetic pathway. The student will determine the structure of these proteins, and will test the wild-type enzyme and selected mutants for enzyme activity. The student will test the activity of the enzymes in recombinant E. coli pathways to confirm the effects of mutants. The student will particularly look for enzyme variants that display altered activity that might be valuable in the whole-cell context. The student will also work with modelling software to study the enzymes using molecular dynamics and quantum mechanical simulations to support proposed enzymatic mechanisms. These data will then be used to model drug binding to the proteins as a first step towards drug development.The student's work will demonstrate the mechanisms of the novel enzymes in the O-antigen pathway. The student's work will also contribute to the development of recombinant O-antigen for vaccine development and the first stages of drug discovery. This work will therefore have both academic and practical outputs.

贝氏柯克斯体是Q热的病原体，Q热是一种人畜共患疾病，在全球范围内流行，特别是在英国西南部。C.伯内特氏菌是反刍动物的一种专性病原体：在牲畜中，它会导致流产和死胎，造成明显的经济后果。C.贝氏体是人类的机会致病菌，感染剂量非常低。Q热会引起严重的流感样症状。虽然这些通常是自行解决的，但有些患者需要住院治疗，其中一些患者继续遭受严重的并发症（例如心肌炎）。C.贝氏菌是中东地区英国军人所患的所谓“沙漠热”的病原体之一，并与慢性疲劳综合征有关。显然需要有效的治疗C。伯内特氏菌我们是一个正在进行的（BBSRC资助）计划的一部分，使用C的多糖O抗原生产疫苗。Burnetii，目前最好的候选疫苗。这种O抗原含有两种非常不寻常的糖（病毒糖和二氢羟基链霉菌糖）。我们已经确定了几个生物合成的蛋白质，支持我们的假设的生物合成。这些酶显示出至少两种新的酶活性，这可能为制备特异性抑制剂提供了极好的机会。我们有大量制备蛋白质的方案，这应该足以确定蛋白质结构。主要导师将与Diamond Light Source合作，使用即将推出的微电子衍射方法作为该结构解决方案的一部分。在这个项目中，学生将从生物合成途径中选择最具科学意义的蛋白质。学生将确定这些蛋白质的结构，并将测试野生型酶和选定的突变体的酶活性。学生将测试重组大肠杆菌中酶的活性。大肠杆菌途径，以确认突变体的影响。学生将特别寻找在全细胞环境中可能有价值的显示改变活性的酶变体。学生还将使用建模软件研究酶，使用分子动力学和量子力学模拟来支持拟议的酶机制。这些数据将被用来模拟药物与蛋白质的结合，作为药物开发的第一步。学生的工作将展示O-抗原途径中新酶的机制。学生的工作也将有助于开发重组O抗原的疫苗开发和药物发现的第一阶段。因此，这项工作将具有学术和实践成果。