BioEmPiRe; Accessing uncharted but essential landscapes to biological machineries by pulse EPR

为了在分子细节上理解生物过程，人们需要捕捉并可视化起作用的蛋白质。现代结构生物学方法如X射线结晶学和低温电子显微镜可以提供蛋白质结构的静态高分辨率快照，但它们无法捕捉运动中的蛋白质来揭示动态功能。利兹最先进的电子顺磁共振(EPR)光谱仪将能够提取蛋白质位点上设计的一对未配对电子之间的准确距离，从而充当分子纳米尺子。因此，在蛋白质的功能周期中准确测量这种距离将有助于阐明基本的生物过程。脉冲电子顺磁共振是现代生物分子研究中的一种强有力的方法，在过去的10年里取得了巨大的技术进步，灵敏度提高了一个数量级以上。作为利兹的蛋白质结构分子生物学家网络(包括几位领先的EPR专家)，PELDOR被认为是未来生物科学的关键方法。我们在利兹拥有非常庞大的用户基础，全国(帝国、国王、格拉斯哥、圣安德鲁斯)和国外(欧盟、澳大利亚和印度)，以及无与伦比的基础生物系统，具有代表性的蛋白质横跨所有生命王国。这些蛋白质参与了从癌症和神经退行性变到抗菌素耐药性和新陈代谢等广泛的疾病相关生物学机制。新的EPR光谱仪将首先揭示分子细节的基本生物机制的新信息，这些信息目前无法通过其他方法获得。我们的调查人员、合作者和行业合作伙伴来自广泛的国家和国际机构。我们在EPR以及生物和医学领域有着广泛的记录，预计这一安装将大大提高英国在全球生物EPR方面的能力和声誉。我们的商业案例将确保利兹中心的可持续性，并将服务于东北部和其他大学，正如我们积极支持BioEmPiRe的团体和调查人员名单所表明的那样。通过利兹大学的捐款，将获得一名EPR工作人员科学家的职位，最初为期两年。此外，还将对预定地点进行全面翻新，并购买一台冷冻机，以实现光谱仪的最佳安装和运行。该工具将成为英国学术和工业网络的一部分，进一步确保可持续性。这些升级将使英国保持国际竞争力，并继续开发和应用EPR方法来解决生物科学中的重要问题。

To understand biological processes at a molecular detail one needs to capture and visualize proteins in action. Modern structural biology methods such as X-ray crystallography and cryo-electron microscopy (CryoEM) could provide static high-resolution snapshots of protein structures, but they are unable to capture proteins in motion to reveal dynamic function. The new state-of-the-art electron paramagnetic resonance (EPR) spectrometer in Leeds will be capable of extracting accurate distances between pairs of unpaired electrons engineered on protein sites and thus acting as a molecular nanoscale-ruler. The accurate measurement of such distances over a protein's functional cycle will thereby enable the elucidation of fundamental biological processes. Pulsed EPR is a powerful method in modern biomolecular research and has seen tremendous technical advances over the last 10 years with sensitivity increasing by more than an order of magnitude. As a network of protein structural molecular biologists in Leeds (including several leading EPR specialists), consider PELDOR as a key approach in the future of biosciences. We have a very large base of users in Leeds, nationally (Imperial, King's, Glasgow, St Andrews) and abroad (EU, Australia and India) and an unmatched variety of fundamental biological systems with representative proteins across all kingdoms of life. These proteins are involved in a wide range of disease-related biological mechanisms from cancer and neurodegeneration to antimicrobial resistance and metabolism. Novel information for fundamental biological machineries in molecular detail and currently inaccessible by other methods, would be first revealed by the new EPR spectrometer. Our investigators, collaborators and industrial partners come from a wide range of national and international institutions. We have an extensive track record in the field of EPR and biological and medical sciences and anticipate this installation will substantially increase the UK's capability and reputation in biological EPR worldwide. Our business case will ensure sustainability for the Leeds-based centre and will serve the North East and other Universities as demonstrated by our list of groups and investigators actively supporting BioEmPiRe. The position of an EPR staff scientist will be secured for an initial period of two years through a contribution by the University of Leeds. In addition, intended location will be fully refurbished and a chiller will be purchased to enable the optimal installation and operation of the spectrometer. The instrument will be part of the UK academic and industrial networks further ensuring sustainability. These upgrades will allow the UK to remain internationally competitive and to continue developing and applying the EPR methodology to important problems across the biosciences.