Centre for structural analysis of complex biological systems

复杂生物系统结构分析中心

• 批准号: BB/M012107/1
• 负责人: Ian Collinson
• 金额: $ 69.72万
• 依托单位: University of Bristol
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2014
• 资助国家: 英国
• 起止时间: 2014 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FM012107%2F1
• 关键词: Centre; structural; analysis; complex; biological

Understanding the function of the molecules of life requires knowledge of their three dimensional structures. Seeing, at or near to the level of individual atoms, how the building blocks of life (proteins and DNA) are assembled enables us to understand both how they may act to drive the chemical reactions that power and maintain living cells, and how they are organised into more complex structures that form the basis of cells and tissues. Detailed knowledge of structure can explain how specific alterations affect function, for example where changes to specific molecules are linked to disease, or how biological systems can be engineered to fulfil useful functions, such as making new drugs or turning carbon dioxide into liquid fuels.Most structures of biological molecules are derived from experiments where ordered crystals of the pure material are exposed to X-rays. The success of this approach relies upon inducing crystals to form. Unfortunately, for many interesting and important biological molecules this remains very difficult, and large numbers of experiments must be conducted to identify suitable conditions for crystal formation. However, recent technological developments have increased the number of experiments possible with limited amounts of material, and created automated systems to monitor the progress of experiments and detect crystals as they form. Furthermore, technology has improved our ability to create conditions mimicking those existing inside biological membranes (the structures that separate the cell interior from its surroundings and organise the cell into compartments) greatly simplifying the process of obtaining crystals of proteins that are normally associated with membranes. Such proteins perform key biological functions at the cell surface, enabling cells to recognise one another and to bind biological surfaces, and regulating the traffic of molecules, including other proteins, into and out of the cell. However, membrane proteins are much harder to work with, and hence less well understood, than other protein systems.Here we request funds to purchase equipment that will transform our ability to grow crystals, and obtain structures, of a range of biologically interesting but technically challenging targets. We will create a state-of-the-art Facility to exploit recent successes producing proteins and protein assemblies in the quantities necessary for crystallisation. Specifically, we wish to purchase: i) a robot to set up crystallisation experiments in conditions replicating the membrane environment; ii) an automated system to house the numbers of crystallisation experiments made possible by robotic systems working on small scales, and that will monitor their progress without human intervention; and iii) a complete crystallisation facility, including a robot to set up experiments and a microscope to inspect the results, maintained in a controlled, oxygen-free, environment. We will use this equipment to obtain structures of a number of biological molecules and assemblies including: the machinery controlling protein movement across membranes; the surface proteins of the human red blood cell that determine blood group, surface proteins from disease-causing bacteria that enable them to bind human cells; giant molecular machines synthesising drugs and antibiotics; the protein assembly by which cells carry out the instructions contained within genes; artificial proteins that carry electrons; and a wide range of proteins, involved in processes from bacterial antibiotic resistance to conversion of carbon dioxide into liquid fuels, that only function when oxygen is absent. Through our strong links to other local Universities our Facility, which will be unique within the region, will be open to researchers across the South West and South Wales, and will provide cutting edge instrumentation on which to provide the next generation of scientists with skills essential to the UK science and technology base.

了解生命分子的功能需要了解它们的三维结构。在单个原子水平或接近单个原子的水平上，观察生命的组成部分(蛋白质和DNA)是如何组装的，使我们能够了解它们如何推动为活细胞提供动力和维持活细胞的化学反应，以及它们是如何组织成更复杂的结构，形成细胞和组织的基础。对结构的详细了解可以解释特定的变化是如何影响功能的，例如，特定分子的变化与疾病有关，或者生物系统如何被设计来实现有用的功能，如制造新药或将二氧化碳转化为液体燃料。生物分子结构的大多数来自于将纯材料的有序晶体暴露在X射线下的实验。这种方法的成功依赖于诱导晶体的形成。不幸的是，对于许多有趣和重要的生物分子来说，这仍然是非常困难的，必须进行大量的实验来确定适合晶体形成的条件。然而，最近的技术发展增加了用有限的材料进行实验的次数，并创建了自动化系统来监测实验的进展并在晶体形成时进行检测。此外，技术提高了我们创造模拟生物膜内现有条件的能力，生物膜是将细胞内部与环境分开并将细胞组织成隔间的结构，极大地简化了获得通常与膜相关的蛋白质晶体的过程。这些蛋白质在细胞表面执行关键的生物功能，使细胞能够相互识别并结合生物表面，并调节包括其他蛋白质在内的分子进出细胞的交通。然而，膜蛋白比其他蛋白质系统更难处理，因此了解得更少。在这里，我们请求资金购买设备，这些设备将改变我们生长晶体的能力，并获得一系列具有生物学意义但具有技术挑战性的靶点的结构。我们将创建一个最先进的设施，以利用最近成功地生产结晶所需数量的蛋白质和蛋白质组件。具体地说，我们希望购买：i)一个机器人，用于在复制膜环境的条件下进行结晶实验；ii)一个自动化系统，用于存储小规模工作的机器人系统进行的大量结晶实验，并将在没有人工干预的情况下监控其进展；iii)一个完整的结晶设施，包括一个设置实验的机器人和一个在受控、无氧环境中检查结果的显微镜。我们将使用这台设备来获得一些生物分子和组件的结构，包括：控制蛋白质跨膜移动的机制；决定血型的人类红细胞表面蛋白；来自致病细菌的使它们能够结合人类细胞的表面蛋白；合成药物和抗生素的巨型分子机器；细胞执行基因内指令的蛋白质组装；携带电子的人造蛋白质；以及广泛的蛋白质，参与从细菌抗药性到将二氧化碳转化为液体燃料的过程，这些蛋白质只有在没有氧气的情况下才能发挥作用。通过我们与当地其他大学的紧密联系，我们的设施在该地区将是独一无二的，将向西南和南威尔士的研究人员开放，并将提供尖端仪器，为下一代科学家提供对英国科学和技术基础至关重要的技能。

项目成果

Resistance to the "last resort" antibiotic colistin: a single-zinc mechanism for phosphointermediate formation in MCR enzymes.

• DOI: 10.1039/d0cc02520h
• 发表时间: 2020-05
• 期刊: Chemical communications
• 影响因子: 4.9
• 作者: Emily Lythell;R. Suardíaz;P. Hinchliffe;Chonnikan Hanpaibool;Surawit Visitsatthawong;Sofia Oliveira;Eric J. M. Lang;Panida Surawatanawong;V. Lee;T. Rungrotmongkol;Natalie Fey;J. Spencer;A. Mulholland

The Role of Cytochrome P450 AbyV in the Final Stages of Abyssomicin C Biosynthesis

细胞色素 P450 AbyV 在 Abyssomicin C 生物合成最后阶段的作用

• DOI: 10.1002/ange.202213053
• 发表时间: 2022
• 期刊: Angewandte Chemie
• 作者: Devine A