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(共享仪器拨款)项目申请NIH资金对现有设备进行重大升级 和全功能布鲁克生物自旋时间域E580电子顺磁共振(EPR) 配置为在X-(9.5 GHz)和Q-波段(34 GHz)共振频率下工作的光谱仪以及 该公司于2009年收购。拟议的升级将显著扩展现有的 用于获得#年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中的自由基中间体，以及结构 组织寡核苷酸、水溶性蛋白和膜蛋白复合体。具体的 拟议升级的组件包括增加(1)脉冲整形和频率啁啾 AWG(任意波形发生器)技术支持的功能，(2)300 W Q频段(34 GHz) 放大器进一步实现激励带宽增加5.5倍，以及(3)无制冷剂 变温(5K至300K)附件，以便在最佳状态下进行这些EPR实验 并消除对供应短缺造成的不可再生液氦的依赖 这是这项研究的一个主要瓶颈。升级后的仪器将可以进行时域EPR 在较低的蛋白质浓度下进行实验，增加了距离范围并提高了准确性 更远距离限制和扩展蛋白质系统，特别是膜的研究 由于设备的限制，目前我们对蛋白质复合体的研究存在重大困难。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、南卡罗来纳州和美国的其他合作组织。