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

埃塞克斯大学多用户生物医学 EPR 设施的替换 EPR 光谱仪

• 批准号: BB/T01802X/1
• 负责人: Dimitri Svistunenko
• 金额: $ 46.98万
• 依托单位: University of Essex
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2021
• 资助国家: 英国
• 起止时间: 2021 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FT01802X%2F1
• 关键词: Replacement; EPR; spectrometer; multiuser; Biomedical

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.

我们建议更换目前的电子顺磁共振（EPR）光谱仪，在生物医学EPR设施在埃塞克斯大学，与最新的最先进的X波段光谱仪的能力，为未来upgradations.EPR光谱允许检测的分子的状态，称为顺磁性，形成在所有的氧化/还原反应。这些顺磁状态，其中最著名的是自由基，可以是短暂的（具有短或长的寿命）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过程非常有趣，因为一种类型的顺磁中心转变为另一种类型的顺磁中心的顺序事件可以及时跟踪，从而提供了揭示酶如何工作的分子机制的图片。目前的EMX EPR光谱仪（布鲁克公司）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设施为英国一系列研究小组进行的蛋白质和酶研究做出了重大贡献。该设施在基于蛋白质的自由基和金属蛋白质和酶方面发展了高度专业化的专门知识。我们内部制造的用于酶和底物之间快速反应的EPR光谱的冷冻样品的装置（由BBSRC资助，BB/E02355 X/1）允许获得与顺磁性物质在反应时间过程中的出现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、赠款奖励和员工培训中记录的高质量输出，我们需要一台类似级别的EPR光谱仪（E500 Bruker），但与当前型号相比具有显著优势，即：a）最新的电子和软件包; B）该系统可以在将来被升级以在第二工作频率和脉冲状态下操作。申请人财团由来自埃塞克斯大学两个学院的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出色的国际声誉。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。该提案包括10个项目，这些项目需要并将大大受益于所需仪器的功能：1.探索变铅青链霉菌酶的生物技术潜力2.蛋白质之间的电子传递。温度跃变时温敏智能材料的相变动力学4。金属蛋白质的时间分辨EPR和时间分辨晶体结构的集成5.酶和蛋白质EPR谱的深度神经网络分析6.基于聚合物的脂质颗粒中膜蛋白的动力学7.细菌铁蛋白的新生物化学。新生血红素过氧化物酶9.饮食缺铁与心肌组织呼吸10.来自链霉素的新型转运蛋白样膜蛋白位于埃塞克斯的生物医学EPR设施中现有的布鲁克EMX光谱仪一直是制作高质量出版物和获得资助奖项的关键工具，在设施存在期间，资金和研究产出呈明显上升趋势。更换后的光谱仪将确保我们在生物分子EPR光谱学领域的世界领先地位。

项目成果

Rigidifying a De Novo Enzyme Increases Activity and Induces a Negative Activation Heat Capacity.

• DOI: 10.1021/acscatal.1c01776
• 发表时间: 2021-09-17
• 期刊: ACS catalysis
• 影响因子: 12.9
• 作者: Hindson SA;Bunzel HA;Frank B;Svistunenko DA;Williams C;van der Kamp MW;Mulholland AJ;Pudney CR;Anderson JLR
• 通讯作者: Anderson JLR

EPR spectroscopy of whole blood and blood components: can we diagnose abnormalities?

• DOI: 10.7555/jbr.35.20210011
• 发表时间: 2021-04-30
• 期刊: Journal of biomedical research
• 影响因子: 2.3
• 作者: Svistunenko DA

The circularly permuted globin domain of Androglobin

雄珠蛋白的循环排列珠蛋白结构域

• DOI: 10.21203/rs.3.rs-1685645/v1
• 发表时间: 2022
• 作者: Reeder B