An ODNP Probe to Study Hydration Dynamics in Membrane Protein

用于研究膜蛋白水合动力学的 ODNP 探针

• 批准号: 9896838
• 负责人: Thorsten Maly
• 金额: $ 54.79万
• 依托单位: BRIDGE 12 TECHNOLOGIES, INC.
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-30 至 2021-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9896838
• 关键词: Alzheimer' s Disease; Area; Cell Nucleus; Cell physiology; Communities; Cost Savings; Data; Defect; Development; Disease; Educational process of instructing; Electron Spin Resonance Spectroscopy; Electrons; Ensure; Equipment; Frequencies; Funding; Generations; Health; Heart; Heating; Home environment; Hydration status; Laboratories; Liquid substance; Magnetic Resonance; Maps; Mechanics; Membrane; Membrane Lipids; Membrane Proteins; Methods; NMR Spectroscopy; Nuclear; Output; Parkinson Disease; Performance; Phase; Physiologic pulse; Physiological Processes; Play; Power Sources; Price; Proliferating; Protein Dynamics; Proteins; Research; Research Activity; Research Personnel; Resolution; Resources; Role; Sampling; Scientist; Signal Transduction; Site; Small Business Innovation Research Grant; Solid; Source; Spectrum Analysis; Spin Labels; System; Techniques; Time; United States National Institutes of Health; Water; X-Ray Crystallography; base; computerized data processing; cost effective; design; electron diffraction; experimental study; improved; instrument; instrumentation; interest; irradiation; macromolecule; microwave electromagnetic radiation; programs; protein structure; solid state nuclear magnetic resonance; structural biology; success; technology development; three dimensional structure; usability

Project Summary / Abstract The proposed research focuses on the development of turn-key instrumentation for liquid-state Overhauser Dynamic Nuclear Polarization (ODNP) spectroscopy to study the site-specific translational dynamics of water molecules located at the interface of bio-macromolecules such as membrane proteins. It will allow researchers to readily perform ODNP experiments in a state-of-the-art commercially available X-band cw/pulsed electron paramagnetic resonance (EPR) spectrometer or, alternatively, in a standalone, benchtop ODNP system. In recent years, DNP has proven to be a robust method to increase signal intensities in NMR experiments in laboratories around the world and substantial progress has been made in adapting DNP for solid- and solution- state NMR spectroscopy. This progress has sparked a new interest in ODNP spectroscopy. Although the method is known since the 1960s it has just recently been applied successfully to study the site-specific translational dynamics of water located at the interface of large bio-macromolecules such as membrane proteins. ODNP can map out the local and site-specific hydration dynamics landscape of membrane proteins and lipid membranes and can provide critical information about the protein structure and dynamics. One of the major challenge of this method is that currently no turn-key, robust instrumentation is available. In this SBIR phase 2 application we will build on the phase 1 success. We propose to develop a continuous- wave ODNP resonator, a microwave power source with integrated NMR spectrometer, and an application package for OpenVnmrJ to completely control the experiment. The resonator will have a large conversion factor to reduce the required microwave power and therefore sample heating. The successful development of this technology will provide researchers access to instrumentation allowing them to incorporate ODNP spectroscopy in their research routine without the hassle of troubleshooting home-built equipment. 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 拟议的研究重点是开发交钥匙仪表的液态 Overhauser动态核极化（ODNP）光谱研究位点特异性翻译动力学 水分子位于生物大分子如膜蛋白的界面。它将允许 研究人员可以在最先进的商用X波段cw/pulsed 电子顺磁共振（EPR）光谱仪或可替代地，在独立的台式ODNP系统中。 近年来，DNP已被证明是一种强大的方法，以增加核磁共振实验中的信号强度 在世界各地的实验室中，在使DNP适用于固体和溶液方面取得了实质性进展， 核磁共振谱。这一进展引发了对ODNP光谱学的新兴趣。尽管方法 自20世纪60年代以来，它已被成功地应用于研究位点特异性翻译 水的动力学位于大的生物大分子如膜蛋白的界面。ODNP可以 绘制出膜蛋白和脂质膜的局部和特定位点的水合动力学景观 并且可以提供关于蛋白质结构和动力学的关键信息。 这种方法的主要挑战之一是目前没有交钥匙，强大的仪器。 在此SBIR第2阶段应用中，我们将在第1阶段成功的基础上进行开发。我们建议发展一个持续的- 波ODNP谐振器、集成NMR谱仪的微波功率源及其应用 OpenVnmrJ软件包来完全控制实验。谐振器将具有大的转换因子 以减少所需的微波功率并因此减少样品加热。 这项技术的成功开发将为研究人员提供仪器 使他们能够将ODNP光谱学纳入其研究程序，而无需麻烦地排除故障 自制设备这将极大地推广该方法，并对许多由政府资助的项目产生很大的兴趣。 美国国立卫生研究院