Benchtop Q-Band Pulsed EPR Spectrometer for Intermolecular Distance Measurements

用于分子间距离测量的台式 Q 波段脉冲 EPR 光谱仪

• 批准号: 10484084
• 负责人: Thorsten Maly
• 金额: $ 101.41万
• 依托单位: BRIDGE 12 TECHNOLOGIES, INC.
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-10 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10484084
• 关键词: Alzheimer' s Disease; Cell physiology; Complex; Computers; Cryoelectron Microscopy; Defect; Development; Disease; Electron Spin Resonance Spectroscopy; Electrons; Evaluation; Extravasation; Feedback; Frequencies; Funding; Generations; Goals; Hand; Health; Heart; Length; Liquid substance; Magnetic Resonance Spectroscopy; Measurement; Measures; Membrane Proteins; Methods; Morphologic artifacts; NMR Spectroscopy; Non-Insulin-Dependent Diabetes Mellitus; Parkinson Disease; Performance; Phase; Physiologic pulse; Physiological Processes; Play; Problem Solving; Proliferating; Protein Dynamics; Proteins; Research; Research Personnel; Resolution; Resources; Role; Sampling; Scientist; Shapes; Signal Transduction; Site; Small Business Innovation Research Grant; Source; Spectrum Analysis; Spin Labels; Standardization; Structure; System; Techniques; Temperature; Testing; Time; Training; United States National Institutes of Health; X-Ray Crystallography; base; cost effective; cryostat; design; expectation; experimental study; improved; instrument; instrumentation; interest; macromolecule; operation; phase 1 designs; prevent; protein structure; prototype; structural biology; success; three dimensional structure; tool; vector

Project Summary / Abstract Complex bio-macromolecules such as membrane proteins play crucial roles in many cellular and physiological processes and specific defects are associated with many known. Determining their three- dimensional structures is one of the main objectives in structural biology and the NIH devotes considerable resources towards this goal. In recent years, pulse EPR spectroscopy, in particular Double Electron-Electron Resonance (DEER) has contributed valuable structural constraints to solve structures of bio-macromolecules. However, currently these methods are only available to experts in the field because operating the complex instrument requires lots of training making it difficult for a non-EPR expert to access this highly valuable information. State-of-the-art spectrometer configurations may be the most versatile option; however, many features of current commercially available systems are unnecessary or simply confusing. Options such as continuous wave (cw) EPR are unnecessary when the main objective is to measure distances. In addition, it is also often overwhelming to applications scientist that are not experts in pulsed EPR spectroscopy. Much of the experiment setup and processing can be standardized but current commercial instruments don’t come with these streamlined features. With DEER becoming more popular it is time to propose a solution that is geared specifically towards distance measurements rather than offering a universal research instruments. Removing unnecessary options and focusing only on DEER spectroscopy will allow us to design a compact, turn-key instrument. In this SBIR application we propose to develop a compact, turn-key Q-band pulsed EPR spectrometer for pulsed dipolar spectroscopy. The system will feature a liquid cryogen-free magnet and cryostat, a low-Q but high-sensitivity resonator and a compact, EPR bridge with state-of-the-art pulse shaping capabilities. The system will be fully computer controlled and easy to operate with minimal training. The successful development of this compact, Q-band pulsed EPR spectrometer will provide researchers access to turn-key instrumentation allowing them to easily incorporate experiments such as DEER in their research. This will greatly proliferate the method and is of large interest to many projects funded by the U.S. National Institutes of Health.

项目总结/摘要 复杂的生物大分子，如膜蛋白，在许多细胞和 生理过程和特定缺陷与许多已知的相关。决定他们的三个- 三维结构是结构生物学的主要目标之一，NIH投入了相当大的精力， 为实现这一目标提供资源。近年来，脉冲EPR谱，特别是双电子-电子 共振（DEER）为求解生物大分子结构提供了有价值的结构约束。 然而，目前这些方法仅对该领域的专家可用，因为操作复杂的 仪器需要大量的培训，使非EPR专家难以访问这个非常有价值的 信息. 最先进的光谱仪配置可能是最通用的选择;然而，许多功能 目前商业上可用的系统是不必要的或简单的混淆。连续等选项 当主要目的是测量距离时，波（cw）EPR是不必要的。此外，还经常 这对不是脉冲EPR光谱学专家的应用科学家来说是压倒性的。大部分实验 设置和处理可以标准化，但是当前的商业仪器不具有这些 精简的功能。随着DEER变得越来越受欢迎，是时候提出一个适合 专门用于距离测量，而不是提供通用的研究工具。去除 不必要的选择，只专注于DEER光谱将使我们能够设计一个紧凑的，交钥匙 仪器 在这个SBIR应用中，我们建议开发一个紧凑的，交钥匙的Q波段脉冲EPR光谱仪 用于脉冲偶极光谱。该系统将采用无液体冷冻剂的磁体和低温恒温器，低Q但 高灵敏度谐振器和具有最先进脉冲整形能力的紧凑型EPR电桥。系统 将完全由电脑控制，操作简单，只需很少的培训。 这种紧凑的Q波段脉冲EPR光谱仪的成功开发将为研究人员提供 获得交钥匙仪器，使他们能够轻松地将实验，如鹿在他们的 research.这将极大地推广该方法，并对美国资助的许多项目产生重大影响。 国立卫生研究院。