Upgrade of Bruker Time-domain EPR Spectrometer

布鲁克时域EPR光谱仪升级

• 批准号: 10431412
• 负责人: ALEX I. SMIRNOV
• 金额: $ 59.99万
• 依托单位: NORTH CAROLINA STATE UNIVERSITY RALEIGH
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10431412
• 关键词: Amplifiers; Back; Biochemical; Dependence; Electron Spin Resonance Spectroscopy; Enzymes; Equipment; Free Radicals; Frequencies; Funding; Grant; Helium; Liquid substance; Membrane Proteins; Metals; Minor; North Carolina; Oligonucleotides; Physiologic pulse; Process; Proteins; Research; Shapes; South Carolina; System; Technology; Temperature; Time; United States National Institutes of Health; Universities; Water; experimental study; improved; instrument; instrumentation; protein complex

This SIG (Shared Instrumentation Grant) project requests NIH funds for a major upgrade of an existing and fully functional Bruker Biospin time domain E580 electron paramagnetic resonance (EPR) spectrometer configured to operate at X- (9.5 GHz) and Q-band (34 GHz) resonance frequencies and acquired back in 2009. The proposed upgrade will significantly expand capabilities of the existing instrument for obtaining atomic-level details on the structural organization of catalytic metal centers in enzymes, free radical intermediates of biochemical and photosynthetic processes, and structural organization oligonucleotides, water soluble proteins, and membrane protein complexes. The specific components of the proposed upgrades include adding (1) pulse-shaping and frequency chirping capabilities enabled by AWG (arbitrary waveform generator) technology, (2) 300 W Q-band (34 GHz) amplifier to achieve a further - up to 5.5-fold - increase in the excitation bandwidth, and (3) cryogen-free variable temperature (5 K to 300 K) accessory to carry out these EPR experiments at optimal temperatures and eliminate dependence on a non-renewable liquid Helium which short supply is creating a major bottleneck in the research. The upgraded instrument will allow for carrying out time-domain EPR experiments at lower protein concentrations, increase the distance range and improve accuracy of longer-range distance constrains, and expand studies of protein systems and, particularly, membrane protein complexes, we have currently major difficulties with because of the equipment limitations. These capabilities will benefit a group if six Major and four Minor users from North Carolina State University and other collaborating groups in North and South Carolina and the USA.

该 SIG（共享仪器补助金）项目请求 NIH 提供资金，对现有仪器进行重大升级 和功能齐全的 Bruker Biospin 时域 E580 电子顺磁共振 (EPR) 配置为在 X 波段 (9.5 GHz) 和 Q 波段 (34 GHz) 谐振频率下运行的光谱仪 于 2009 年收购。拟议的升级将显着扩展现有的功能 用于获得催化金属中心结构组织的原子级详细信息的仪器 酶、生化和光合作用过程的自由基中间体以及结构 组织寡核苷酸、水溶性蛋白质和膜蛋白复合物。具体的 拟议升级的组成部分包括添加 (1) 脉冲整形和频率啁啾 由 AWG（任意波形发生器）技术实现的功能，(2) 300 W Q 频段 (34 GHz) 放大器以实现激励带宽进一步增加高达 5.5 倍，以及 (3) 无制冷剂 可变温度（5 K 至 300 K）附件，以最佳状态进行这些 EPR 实验 温度并消除对因供应短缺而造成的不可再生液氦的依赖 研究的一大瓶颈。升级后的仪器将允许执行时域 EPR 在较低蛋白质浓度下进行实验，增加距离范围并提高准确性 更长距离的距离限制，并扩大了蛋白质系统的研究，特别是膜 由于设备的限制，我们目前在蛋白质复合物方面遇到了很大的困难。这些 如果来自北卡罗来纳州立大学的六名主要用户和四名次要用户以及 北卡罗来纳州、南卡罗来纳州以及美国的其他合作团体。