Centre for structural analysis of complex biological systems

理解生命分子的功能需要了解它们的三维结构。在单个原子或接近单个原子的水平上观察生命的构建块（蛋白质和DNA）是如何组装的，使我们能够理解它们如何驱动化学反应，从而为活细胞提供动力和维持，以及它们如何被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改变如何影响功能，例如，特定分子的变化与疾病有关，或者如何设计生物系统以实现有用的功能，例如制造新药或将二氧化碳转化为液体燃料。大多数生物分子的结构都是从纯物质的有序晶体暴露在x射线下的实验中得到的。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晶体形成的合适条件。然而，最近的技术发展增加了用有限材料进行实验的可能性，并创造了自动化系统来监测实验的进展和检测晶体的形成。此外，技术已经提高了我们创造模拟生物膜（将细胞内部与周围环境分离并将细胞组织成隔室的结构）内部条件的能力，大大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学功能，使细胞能够相互识别并结合生物表面，并调节分子（包括其他蛋白质）进出细胞的流量。然而，与其他蛋白质系统相比，膜蛋白更难处理，因此对其了解较少。在此，我们请求资金来购买设备，以改变我们培养晶体的能力，并获得一系列生物学上有趣但技术上具有挑战性的目标的结构。我们将建立一个最先进的设施，利用最近的成功生产蛋白质和蛋白质组装，以达到结晶所需的数量。具体来说，我们希望购买：i)一个机器人，在复制膜环境的条件下进行结晶实验；Ii)一个自动化系统，用于容纳由机器人系统在小尺度上工作所实现的结晶实验数量，并在没有人为干预的情况下监测实验进展；iii)一个完整的结晶设备，包括一个机器人来设置实验和一个显微镜来检查结果，保持在一个可控的，无氧的环境中。我们将使用该设备获得许多生物分子和组件的结构，包括：控制蛋白质跨膜运动的机制；人类红细胞的表面蛋白质决定血型，致病细菌的表面蛋白质使它们能够结合人类细胞；巨大的分子机器合成药物和抗生素；细胞执行基因中包含的指令的蛋白质组合；携带电子的人造蛋白质；从细菌对抗生素的耐药性到将二氧化碳转化为液体燃料，各种各样的蛋白质只有在缺氧的情况下才能发挥作用。通过我们与其他当地大学的紧密联系，我们的设施将在该地区独一无二，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。

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.