Applications of NMR Spectroscopy to Study Structure, Dynamics and Small Molecule Interactions Related to Protein Folding and Misfolding

核磁共振波谱是研究生物大分子在自然环境中的结构、动力学和相互作用的有力技术，包括蛋白质和细胞环境，以及在固体环境中的生物膜、生物材料和膜蛋白。这种方法使研究人员能够获得这些成分的详细原子分辨率图像，并研究这些生物分子产生的相互作用的强度和频率，从而获得感兴趣的系统的全面图像。溶液生物分子核磁共振的一个独特特征，特别是最近开发的TXO冷冻探针，是表征蛋白质的残余结构和瞬态相互作用的能力，这些蛋白质缺乏明显的三维结构，被称为内在无序。内在紊乱是真核生物基因组编码蛋白质的主要特征，这些蛋白质是许多细胞过程以及异常人类疾病（包括癌症、神经退行性疾病、扩张型心肌病）的核心。生物分子核磁共振的另一个重点研究领域是在固态内；由于自组装或与生物膜或蛋白质网络的相互作用，样品处于更刚性的构象。生物分子ssNMR已经推动了蛋白质淀粉样原纤维和膜蛋白的表征，并且是一种非常准确的方法来探测生物分子组件的结构特性，即生物材料，大蛋白复合物和水凝胶。化学系的核磁共振设备一直服务于剑桥地区的大型分子科学家社区，包括化学系，错误折叠疾病中心（CMD）以及一些公司和其他部门。申请人的研究实验室最近的研究进展使蛋白质聚集和错误折叠疾病领域的重大发现成为可能，为此，CMD已获得资助，以促进过渡研究，以寻找治疗神经退行性疾病，包括阿尔茨海默病和帕金森病。CMD的活动，以及使用我们的核磁共振设备的其他研究实验室，将极大地受益于高通量筛选与这些疾病相关的前体蛋白的小分子。尽管我们的旗舰700 MHz仪器配备了冷冻探针TXO和样品交换器，但这种方法目前无法有效应用，我们需要更现代化的光谱仪控制台来提高灵敏度并应用快速数据采集方法。更换控制台是我们更新700 MHz光谱仪的更大计划的一部分，包括对磁铁的维护，以确保对仪器进行更经济有效的管理，并避免未来的损坏。除了我们在溶液核磁共振方面的专业知识外，我们还在研究与阿尔茨海默病和帕金森病相关的聚集物种方面获得了国际相关的专业知识。在这种情况下，我们希望为我们的核磁共振设备配备生物固体核磁共振能力，用于蛋白质研究，这在剑桥地区令人惊讶地缺失。该设备还将允许对与阿尔茨海默病和帕金森病相关的特定蛋白质聚集的选定分子进行药物筛选。除了该应用程序将对与神经退行性疾病中内在无序蛋白质和蛋白质聚集研究相关的CMD活动带来巨大好处之外，新的NMR功能将为剑桥大学的科学家和经常访问该设施的合作机构（包括工业合作者）提供更大的服务。促进和影响有机化学、材料科学和化学生物学各个方面的研究。

NMR spectroscopy is a powerful technique to study the structure, dynamics and interactions of biomacromolecules in their natural environment in solution, including proteins the cellular milieu, and in the solid-state, including biological membranes, biomaterials and membrane-proteins. This methodology allows researchers to gain detailed atomic resolution pictures of these components and to also study the strength and frequency of the interactions incurred by these biomolecules, giving a comprehensive picture of the systems of interest. A unique feature of the solution biomolecular NMR, in particular recently developed TXO cryoprobes, is the ability to characterise residual structure and transient interactions by proteins which lack distinct three dimensional structures and which are referred to as intrinsically disordered. Intrinsic disorder is a primary characteristic of ~30% of the proteins encoded in the eukaryotic genome and these proteins are central to a number of cellular processes as well as aberrant human diseases including cancer, neurodegenerative disorders, dilated cardiomyopathies. Another key research area in biomolecular NMR is within the solid-state; where samples are in more rigid conformations due to self-assembly or interactions with biological membranes or protein networks. Biomolecular ssNMR has been driving the characterisation of protein amyloid fibrils and membrane proteins, and is an extremely accurate method to probe the structural properties of biomolecular assemblies, i.e.biomaterials, large protein complexes and hydrogels. The NMR facility in the Department of Chemistry has been serving a large community of molecular scientists in the Cambridge area, including the Departments of Chemistry, the Centre for Misfolding Disease (CMD) and a number of companies and other Departments. Recent research developments in the research labs of the applicants have enabled significant discoveries in the field of protein aggregation and misfolding diseases, for which the CMD has been funded to boost the transitional research into finding cures for neurodegenerative disorders include Alzheimer's and Parkinson's diseases. The activities of the CMD, as well as of the other research labs accessing our NMR facility, would greatly benefit from the availability of high-throughput screening of small molecules on the precursor proteins linked with these diseases. This methodology cannot, currently, be applied with efficiency, despite our flagship 700 MHz instrument being equipped with a cryoprobe TXO and sample exchanger, and we need a more modern spectrometer console to boost sensitivity and apply fast methods of data acquisitions. The change of console is part of our bigger plan to rejuvenate our 700 MHz spectrometer, including maintenance to the magnet to ensure more cost effective management of the instrument and avoid future breakage.In addition to our expertise in solution NMR, we have gained internationally relevant expertise in studying the aggregated species that are associated with Alzheimer's and Parkinson's disorders. In this context, we would like to equip our NMR facility with bio-solid NMR capability for protein investigations, which is surprisingly missing in the Cambridge area. This equipment will allow also, the drug-screening of selected molecules to specific protein aggregates associated with Alzheimer's and Parkinson's. In addition to the great benefits that this application would have on the CMD activities associated with the study of intrinsically disordered proteins and protein aggregation in neurodegenerative disorders, the new NMR capability will serve the larger community of scientists at the University of Cambridge and the collaborating institutions that frequently access this facility, including industrial collaborators, faciliting and impacting on research encompassing various aspects of organic chemistry,material sciences and chemical biology.