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: 10010151
• 负责人: Thorsten Maly
• 金额: $ 22.73万
• 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 2022-02-09
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10010151
• 关键词: Alzheimer' s Disease; Cell physiology; Collaborations; Complex; Computers; Coupling; Cryoelectron Microscopy; Defect; Development; Disease; Electromagnetics; Electron Spin Resonance Spectroscopy; Electrons; Elements; Extravasation; Frequencies; Funding; Goals; Hand; Health; Heart; Liquid substance; Magnetic Resonance Spectroscopy; Measurement; Measures; Mechanical Stress; Mechanics; 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; Residual state; Resolution; Resources; Role; Sampling; Scientist; Shapes; Signal Transduction; Site; Small Business Innovation Research Grant; Spectrum Analysis; Spin Labels; Standardization; Structure; System; Techniques; Temperature; Time; Training; United States National Institutes of Health; Variant; X-Ray Crystallography; base; cost effective; cryostat; design; electric field; experimental study; improved; instrument; instrumentation; interest; macromolecule; microwave electromagnetic radiation; operation; prevent; programs; protein structure; prototype; simulation; structural biology; 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 overly complicated. 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. Today, much of the experiment setup and processing can be automated and standardized. Current commercial solutions do not come equipped with these streamlined features. With DEER becoming more popular it is time to propose a solution that is geared specifically towards distance measurements than creating 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 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.

项目概要/摘要 膜蛋白等复杂的生物大分子在许多细胞和生物体中发挥着至关重要的作用。 生理过程和特定缺陷与许多已知的相关。确定他们的三 维度结构是结构生物学的主要目标之一，NIH 投入了大量资金 资源来实现这一目标。近年来，脉冲EPR光谱，特别是双电子-电子 共振（DEER）为解决生物大分子的结构提供了有价值的结构约束。 然而，目前这些方法仅适用于该领域的专家，因为操作复杂 仪器需要大量培训，这使得非 EPR 专家很难获得这一极具价值的工具 信息。 最先进的光谱仪配置可能是最通用的选择，但是，许多功能 当前商用系统的一些部分是不必要的或过于复杂的。选项如连续 当主要目标是测量距离时，波 (cw) EPR 是不必要的。此外，还经常 对于不是脉冲 EPR 光谱学专家的应用科学家来说，这是难以承受的。今天，大部分 实验设置和处理可以自动化和标准化。目前的商业解决方案不 配备了这些简化的功能。随着 DEER 变得越来越流行，是时候提出一个 专门针对距离测量的解决方案，而不是创建通用研究 仪器。删除不必要的选项并仅关注 DEER 光谱将使我们能够设计一个 紧凑的交钥匙仪器。 在此 SBIR 应用中，我们建议开发一款紧凑型交钥匙 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磁体和低温恒温器，这是一种低 Q 但 高灵敏度谐振器和具有最先进脉冲整形功能的紧凑型 EPR 电桥。系统 将完全由计算机控制，并且只需最少的培训即可轻松操作。 这种紧凑型 Q 波段脉冲 EPR 光谱仪的成功开发将为研究人员提供 使用交钥匙仪器使他们能够轻松地将 DEER 等实验纳入他们的实验中 研究。这将极大地推广该方法，并且引起美国资助的许多项目的浓厚兴趣。 美国国立卫生研究院。