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.

这个SIG（共享仪器补助金）项目要求NIH资金对现有的 和功能齐全的Bruker Biospin时域E580电子顺磁共振（EPR） 光谱仪，其被配置为在X-（9.5GHz）和Q-频带（34 GHz）谐振频率下操作，以及 2009年收购的。拟议的升级将大大扩大现有 用于获得催化金属中心的结构组织的原子级细节的仪器， 酶、生物化学和光合作用过程的自由基中间体以及结构 组织寡核苷酸、水溶性蛋白和膜蛋白复合物。具体 拟议中的升级组件包括：增加（1）脉冲整形和频率啁啾 通过AWG（任意波形发生器）技术实现的功能，（2）300 W Q波段（34 GHz） 放大器，以实现激励带宽的进一步增加-高达5.5倍，以及（3）无冷冻剂 可变温度（5 K至300 K）的附件进行这些EPR实验在最佳 温度和消除对不可再生的液态氦的依赖， 研究中的一个主要瓶颈。升级后的仪器将允许进行时域EPR 实验在较低的蛋白质浓度，增加距离范围，提高准确性， 更长距离的限制，并扩大了蛋白质系统的研究，特别是膜 蛋白质复合物，我们目前有很大的困难，因为设备的限制。这些 如果来自北卡罗来纳州州立大学的六个主要用户和四个次要用户， 北、南卡罗来纳州和美国的其他合作团体。