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: 9265623
• 负责人: Thorsten Maly
• 金额: $ 6.18万
• 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 2017-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9265623
• 关键词: Alzheimer' s Disease; Area; Cell Nucleus; Cell physiology; Crystallography; Defect; Development; Disease; Electron Nuclear Double Resonance; Electron Spin Resonance Spectroscopy; Electrons; Equipment; Evaluation; Frequencies; Funding; Health; Heart; Home environment; Hydration status; Laboratories; Liquid substance; Maps; Mechanics; Membrane; Membrane Lipids; Membrane Proteins; Methods; NMR Spectroscopy; Nuclear; Parkinson Disease; Performance; Phase; Physiological Processes; Play; Price; Proliferating; Protein Dynamics; Proteins; Research; Research Activity; Research Personnel; Resolution; Resources; Role; Sampling; Signal Transduction; Site; Small Business Innovation Research Grant; Spectrum Analysis; Spin Labels; Structure; System; Techniques; Temperature; United States National Institutes of Health; Water; base; cost; cost effective; design; electron diffraction; improved; instrumentation; interest; irradiation; macromolecule; programs; protein structure; prototype; public health relevance; research study; solid solution; solid state nuclear magnetic resonance; structural biology; success; technology development; three dimensional structure

 DESCRIPTION (provided by applicant): The proposed research focuses on the development of a cost-effect, turn-key probe 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 electron paramagnetic resonance (EPR) spectrometer. 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 o 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. Currently there is no commercial probe available to readily start 9 GHz ODNP experiments; which means the method is only available to a small number of labs that are tech savvy and willing to tinker with their instrumentation. In this SBIR Phase I application we propose to develop a turn-key, cost-effective ODNP probe to be used in a state-of-the-art continuous wave X-Band EPR spectrometer, which can be found in many academic spectroscopy facilities. The successful development of this technology will provide researchers access 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.

 描述（由申请人提供）：拟议的研究重点是开发一种成本效益高、交钥匙的液态奥豪瑟动态核极化（ODNP）光谱探针，以研究位于它将使研究人员能够在最先进的商用 X 波段电子顺磁共振 (EPR) 中轻松进行 ODNP 实验。近年来，DNP 已被证明是一种在世界各地实验室的 NMR 实验中提高信号强度的可靠方法，并且在将 DNP 用于固态和溶液态 NMR 光谱方面取得了实质性进展。尽管该方法自 20 世纪 60 年代以来就已广为人知，但最近才成功应用于研究位于大型生物大分子等界面处的水的位点特异性平移动力学。作为膜蛋白，ODNP 可以绘制膜蛋白和脂质膜的局部和位点特异性水合动力学景观，并且可以提供有关蛋白质结构和动力学的关键信息。目前没有商业探针可轻松启动 9 GHz ODNP 实验；这意味着该方法仅适用于少数精通技术并愿意修补仪器的实验室。在这个 SBIR 第一阶段应用中，我们建议开发一种交钥匙的、具有成本效益的 ODNP 探针，用于最先进的连续波 X 波段 EPR 光谱仪，可在许多学术光谱设施中找到，该技术的成功开发将使研究人员能够将 ODNP 光谱纳入其研究常规，而无需在家中进行故障排除。这将极大地推广该方法，并且引起美国国立卫生研究院资助的许多项目的极大兴趣。