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

应用核磁共振波谱研究与蛋白质折叠和错误折叠相关的结构、动力学和小分子相互作用

基本信息

批准号：
BB/R013535/1
负责人：
Christopher Dobson
金额：
$ 35.06万
依托单位：
University of Cambridge
依托单位国家：
英国
项目类别：
Research Grant
财政年份：
2018
资助国家：
英国
起止时间：
2018 至无数据
项目状态：
已结题

来源：
https://gtr.ukri.org/projects?ref=BB%2FR013535%2F1
关键词：
Applications NMR Spectroscopy Study Structure

项目摘要

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.

核磁共振波谱是一种强大的技术来研究生物大分子在其自然环境中的结构，动力学和相互作用，在溶液中，包括蛋白质，细胞环境，以及在固态，包括生物膜，生物材料和膜蛋白质。这种方法使研究人员能够获得这些组分的详细原子分辨率图像，并研究这些生物分子相互作用的强度和频率，从而全面了解感兴趣的系统。溶液生物分子NMR，特别是最近开发的TXO冷冻探针的独特特征是能够通过缺乏独特的三维结构并且被称为固有无序的蛋白质来消除残留结构和瞬时相互作用。内源性疾病是真核基因组中编码的约30%的蛋白质的主要特征，并且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、扩张性心肌病）的中心。生物分子NMR的另一个关键研究领域是固态;其中样品由于自组装或与生物膜或蛋白质网络的相互作用而处于更刚性的构象。生物分子ssNMR一直在推动蛋白质淀粉样纤维和膜蛋白的表征，并且是探测生物分子组装体（即生物材料、大蛋白质复合物和水凝胶）的结构特性的极其准确的方法。化学系的核磁共振设备一直为剑桥地区的分子科学家提供服务，包括化学系，错误折叠疾病中心（CMD）和一些公司和其他部门。申请人的研究实验室中的最新研究进展已经在蛋白质聚集和错误折叠疾病领域中实现了重大发现，CMD已经获得资助以促进过渡研究，以找到治疗包括阿尔茨海默病和帕金森病在内的神经退行性疾病的方法。CMD的活动，以及其他研究实验室访问我们的核磁共振设施，将大大受益于高通量筛选与这些疾病相关的前体蛋白的小分子的可用性。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，但目前这种方法无法有效应用，我们需要一个更现代的光谱仪控制台来提高灵敏度并应用快速数据采集方法。更换控制台是我们振兴700 MHz光谱仪的更大计划的一部分，包括维护磁铁，以确保更经济有效地管理仪器并避免未来的损坏。除了我们在溶液NMR方面的专业知识外，我们还在研究与阿尔茨海默氏症和帕金森氏症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NMR设备配备用于蛋白质研究的生物固体NMR能力，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氏症和帕金森氏症相关的特定蛋白质聚集体的选定分子进行药物筛选。除了这项应用将对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活动带来巨大好处外，新的NMR能力将为剑桥大学的更大科学家社区以及经常访问该设施的合作机构提供服务，包括工业合作者，促进和影响研究，包括有机化学，材料科学和化学生物学的各个方面。