Replacement EPR spectrometer to the multiuser Biomedical EPR Facility at the University of Essex

我们建议将埃塞克斯大学生物医学EPR设施现有的电子顺磁共振（EPR）光谱仪替换为最先进的x波段光谱仪，该光谱仪具有未来升级的能力。EPR光谱学允许检测在所有氧化/还原反应中形成的称为顺磁性的状态的分子。这些顺磁状态，其中最著名的是自由基，可以是短暂的（或短或长的寿命）或稳定的。酶和蛋白质中顺磁状态的形成过程以及它们的消失是非常有趣的，因为一种顺磁中心转变为另一种顺磁中心的顺序事件可以及时跟踪，从而提供了一幅揭示酶如何工作的分子机制的图片。这些信息对于理解广泛的生物过程是重要的。目前的EMX EPR光谱仪（Bruker）已经使用了23年，并且出现了严重的间歇性故障。这些年来，生物医学EPR设施对英国一系列研究小组进行的蛋白质和酶的研究做出了重大贡献。该机构在蛋白质自由基、金属蛋白和酶方面发展了高度专业化的专业知识。我们内部建造的冷冻样品设备用于酶和底物之间快速反应的EPR光谱（由BBSRC资助，BB/E02355X/1），可以获得与顺磁性物质在反应时间过程中的外观和转化有关的有价值的动力学数据。这些数据集可用于计算机生成的动力学模型，以验证有关酶机制的假设。为了保持该设施在出版物、赠款奖励和工作人员培训中记录的高质量产出，我们需要一个类似级别的EPR光谱仪（E500 Bruker），但与现有型号相比具有实质性优势，即：a)最新的电子和软件包；B)该系统可以在未来升级为在第二工作频率和脉冲状态下工作。申请人财团由来自埃塞克斯大学两所学院的11名学者和8名外部合作者组成，他们都在各自的领域拥有卓越的国际声誉。这些科学家的研究范围很广，但都需要EPR光谱作为他们的研究方法。该建议包括10个项目，这些项目需要并将从所要求的工具的能力中获益匪浅：探索lividans链霉菌酶的生物技术潜力通过和在蛋白质之间传递电子。温度跳变时热响应智能材料相变动力学[j]。集成时间分辨EPR和金属蛋白的时间分辨晶体结构酶和蛋白质EPR谱的深度神经网络分析[j]。膜蛋白在聚合物基脂质颗粒中的隔离动力学细菌铁蛋白的生物化学新进展新生血红素过氧化物酶。饮食缺铁与心肌组织呼吸在埃塞克斯生物医学EPR设施现有的布鲁克EMX光谱仪一直是生产高质量出版物和获得拨款奖励的关键工具，在设施存在期间，资金和研究产出明显上升。更换光谱仪将确保我们在生物分子EPR光谱方面的世界领先地位将保持下去。

We propose to replace the current Electron Paramagnetic Resonance (EPR) spectrometer, at the Biomedical EPR Facility at the University of Essex, with the latest state of the art X-band spectrometer with capabilities for future upgrades.EPR spectroscopy allows detection of molecules in the states, called paramagnetic, that are formed in all oxidation/reduction reactions. These paramagnetic states, of which the best known are free radicals, can be transient (with either short or long life time) or stable. The processes of paramagnetic states formation within enzymes and proteins, as well as their disappearance, are of great interest since the sequential events of one type of paramagnetic centre transforming into another can be followed in time, thus providing a picture of the molecular mechanism revealing how an enzyme works. Such information is important for understanding a wide range of biological processes.The current EMX EPR spectrometer (Bruker) is 23 years old and has developed a serious intermittent fault. Throughout these years, the Biomedical EPR Facility has contributed significantly to the research focused on proteins and enzymes conducted by a range of research groups in the UK. The Facility has developed highly specialised expertise in protein-based free radicals and metallo-proteins and enzymes. Our in-house built apparatus for freezing samples for EPR spectroscopy of rapid reactions between enzyme and substrate (funded by BBSRC, BB/E02355X/1), allows valuable kinetic data pertaining to the appearance and transformation of paramagnetic species over a reaction time course to be obtained. Such data sets can be used in computer generated kinetic models to verify hypotheses about the enzyme mechanism.To maintain the high quality output of the Facility documented in publications, grant awards and staff training, we require an EPR spectrometer of a similar class (E500 Bruker) but with substantial advantages compared to the current model, namely: a) up-to-date electronics and software package; b) the system can be upgraded in the future to operate at a second working frequency and a pulsed regime.The consortium of applicants consists of 11 academics from two Schools of the University of Essex and 8 external collaborators, all with excellent international reputations in their fields. The scope of research by these scientists is wide but all require EPR spectroscopy as a method for their research.The proposal features 10 projects that require and will benefit greatly from the capabilities of the requested instrument:1. Exploring the biotechnological potential of Streptomyces lividans enzymes2. Electron transfer through and between proteins3. Kinetics of the phase transition in thermoresponsive smart materials on temperature jump4. Integrated time-resolved EPR and time-resolved crystal structures of metalloproteins5. Deep neural network analysis of the EPR spectra of enzymes and proteins6. Dynamics of membrane proteins sequestered in polymer based lipid particles7. New biochemistry of bacterioferritin8. De novo haem peroxidases9. Iron deficiency in diet and cardiac muscle tissue respiration10. New transporter-like membrane protein from StreptomycesThe existing Bruker EMX spectrometer at the Biomedical EPR Facility at Essex has been the key instrument in producing high quality publications and obtaining grant awards with a clear upward trajectory of funds and research outputs over the period of the Facility existence. The replacement spectrometer would ensure that our world leading position in EPR spectroscopy of biomolecules will be maintained.