Deuteration Initiative for Neutron Scattering and NMR Studies of Biological Molecules

本提案要求EPSRC支持一个由英国科学家组成的具有高度生物学意义的研究项目。所有这些都将使用复杂的方法，使部分生物分子的氢原子被其较重的“同位素”氘取代。这允许新的中子散射和核磁共振工作，其中这些部分被突出显示，提供其他方法无法获得的关于结构和运动的重要信息。这种“氘化”或“标记”需要在朗格万研究所的格勒诺布尔氘化实验室（ILL由英国拥有/资助33%）开发的创新方法。该实验室是由EPSRC先前的拨款建造的，该拨款为其提供了设备，并启动了英国推动的科学活动。它现在是世界上独一无二的设施，拥有蓬勃发展的内部和用户程序。位于一个非凡的科学校园，下面描述的基于英国的项目也将受益于其他强大的平台。DNA t束序列的中子研究（Keele）“t束”是胸腺嘧啶重复运行的DNA区域。他们参与了监管程序。在旨在了解这些序列以及它们如何与功能相关的研究中，氘化将用于突出两条DNA链中的每一条。丝状病毒的结构和动力学（Cambridge, Keele）丝状病毒是生物学基本过程的独特模型，在药物发现和疫苗设计中也有重要的技术用途。我们将使用氘化方法来研究病毒的结构和运动。一种机械敏感通道蛋白的核磁共振/中子研究（牛津大学）我们将研究一种在不同环境下控制细胞形状和大小的膜上的“机械敏感”蛋白。我们在制作氘化样品方面取得了突破，因此我们可以获得可能与疾病及其治疗有关的重要信息。多酶复合体的中子散射研究（格拉斯哥）我们将研究如何将一组特定的蛋白质组合在一起，形成一个具有医学重要性的专门的“分子机器”，控制将葡萄糖转化为能量途径的关键步骤。细菌蛋白质-DNA复合物的研究（Portsmouth）所有的细菌物种都有一个“免疫系统”，因此它们自己的DNA可以与外来的DNA区分开来。氘化将使研究对理解如何克服细菌防御具有重要意义。“IMPase”酶的研究，与双相情感障碍（南安普顿）双相情感障碍是一种精神疾病。它通常用锂处理，锂与IMPase酶相互作用。我们将研究其与锂的相互作用，以了解锂疗法的性质、副作用和潜在的替代疗法。DNA和药物-DNA复合物的中子衍射研究（阅读）DNA并不总是人们想象中美丽的双螺旋结构。它形成了许多尚未被正确理解的“氢键”相互作用。抗癌药物和其他dna结合药物会干扰这些相互作用。氘化将使这些效应第一次被看到。核磁共振和中子对染色质结构的研究（剑桥）细胞内的DNA与蛋白质捆绑在一起，形成一种叫做染色质的结构。氘化将用于研究关键蛋白复合物如何调节染色质结构的形成，以及哪些基因被表达（转录成RNA和蛋白质）。人体铁转运蛋白的结构研究（伦敦国王学院）铁转运蛋白是一种参与膳食铁从肠道转运到血液的蛋白质。它是血色素沉着病和地中海贫血治疗干预的主要目标。氘化将用于提供有助于药物设计的信息。

This proposal requests EPSRC support for a consortium of UK scientists for research projects of high biological interest. All will use sophisticated methods that enable parts of biomolecules to have their hydrogen atoms replaced by its heavier 'isotope' deuterium. This allows novel neutron scattering and NMR work in which these parts are highlighted, providing important information on structure and movement that is not available by other methods. This 'deuteration' or 'labelling' requires innovative approaches that have been developed at the Grenoble Deuteration Laboratory at the Institut Laue Langevin (the ILL is 33% UK-owned/funded). The laboratory was built with funding from a previous EPSRC grant that equipped it and initiated UK-driven scientific activity. It is now a unique facility in the world, with a flourishing in-house and user programme. Located on a remarkable science campus, the UK-based projects described below will also benefit from other powerful platforms available.1. Neutron studies of T-tract sequences of DNA (Keele)'T-tracts' are regions of DNA with repetitive runs of thymine. They have been implicated in regulatory processes. Deuteration will be used to highlight each of the two DNA strands in studies aimed at understanding these sequences and how they may be related to function.2. Structure and dynamics in filamentous viruses (Cambridge, Keele) Filamentous viruses are unique models for fundamental processes inbiology, and also have important technological uses in drug discovery and vaccine design. We will use deuteration methods to study structure and movement within the virus3. NMR/neutron studies of a mechanosensitive channel protein (Oxford) We will study a 'mechanosensitive' protein from membranes that controls cell shape and size in different environments. We have made a breakthrough in producing deuterated samples so that we can obtain important information that may be relevant to diseases and their treatment.4. Neutron scattering studies of a multienzyme complex (Glasgow)We will study how a particular set of proteins is put together to make a specialised 'molecular machine' of medical importance that controls a key step in the pathway that converts glucose into energy.5. Studies of bacterial protein-DNA complexes (Portsmouth)All bacterial species have an 'immune system', whereby their own DNA can be distinguished from foreign DNA. Deuteration will allow studies that will be important in understanding how bacterial defences can be overcome.6. Studies of the 'IMPase' enzyme, in relation to bipolar disorder (Southampton)Bipolar disorder is a mental illness. It is often treated with lithium, which interacts with the IMPase enzyme. We will study its interaction with lithium in order to understand the nature of lithium therapy, side effects, and potential alternative therapies.7. Neutron diffraction studies of DNA and drug-DNA complexes (Reading)DNA is not always the beautiful double helix of popular imagination. It forms many 'hydrogen-bonding' interactions that are not properly understood. Anticancer and other DNA-binding drugs interfere with these interactions. Deuteration will allow these effects to be seen for the first time.8. NMR and neutron studies of chromatin structure (Cambridge)DNA within the cell is packaged into bundles with proteins to give a structure called chromatin. Deuteration will be used to study how key protein complexes work to regulate the formation of chromatin structure, and which genes are expressed (transcribed into RNA and protein).9. Structural studies of human ferroportin (King's College London)Ferroportin is a protein that is involved in transferring dietary iron from intestines to blood. It is a prime target for therapeutic intervention in the illnesses haemachromatosis and thalassaemia. Deuteration will be used to provide information that will help drug design.