Development of Biomolecular High Field Pulsed DNP and cwDNP MAS-NMR Applications

生物分子高场脉冲 DNP 和 cwDNP MAS-NMR 应用的开发

• 批准号: 209112340
• 负责人: Professor Dr. Clemens Glaubitz
• 金额: --
• 依托单位: Institut für Physikalische und Theoretische Chemie
• 依托单位国家: 德国
• 项目类别: Major Instrumentation Initiatives
• 财政年份: 2011
• 资助国家: 德国
• 起止时间: 2010-12-31 至 2014-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/209112340?language=en
• 关键词: Development; Biomolecular; Field; Pulsed; DNP

Continuous wave (cw)DNP has been shown to offer unprecedented signal enhancement for solidstate NMR. Exploiting its full potential for biomolecular applications is just at the beginning. Unfortunately, its intrinsic inverse field dependence limits its use to magnetic fields, which are lower, compared to those usually used for biomolecular NMR. In contrast, signal enhancement by pulsed DNP does not depend on the magnetic field but requires further research and development. We propose to establish a pulsed DNP solid-state NMR experiment operating at 263 GHz / 400 MHz to drive such development. A gyrotron equipped with microwave switches, which allows creating coherent microwave pulses, will be integrated into our existing NMR infrastructure. A solid-state NMR double resonance probe with microwave resonator for macroscopically ordered samples will be developed. With this hardware setup, we aim to evaluate different pulsed DNP mechanisms and will apply them to solid-state NMR experiments on oriented and ultra-thin membrane protein samples. We expect that this work will not only allow advancement in the use of ordered samples but also provides us with valuable data based on which pulsed DNP MAS-NMR experiments can be established in the long term. In parallel to this developmental project, we will carry out cwDNP MAS-NMR applications on frozen RNA and protein solutions, membrane protein complexes and complete ribosome particles. Access to this unique research infrastructure will be granted by implementing an open user concept for this research program.

连续波（cw）DNP已被证明为固体核磁共振提供前所未有的信号增强。开发其生物分子应用的全部潜力才刚刚开始。不幸的是，与通常用于生物分子核磁共振的磁场相比，其固有的逆场依赖性限制了其在磁场中的应用。相比之下，脉冲DNP的信号增强不依赖于磁场，但需要进一步的研究和发展。我们建议建立一个263 GHz / 400 MHz的脉冲DNP固态核磁共振实验来推动这一发展。配备微波开关的回旋管，可以产生相干微波脉冲，将集成到我们现有的核磁共振基础设施中。研制了一种用于宏观有序样品的微波谐振腔固体核磁共振双共振探针。有了这个硬件设置，我们的目标是评估不同的脉冲DNP机制，并将其应用于定向和超薄膜蛋白质样品的固态核磁共振实验。我们期望这项工作不仅可以促进有序样品的使用，还可以为我们提供有价值的数据，基于这些数据，可以长期建立脉冲DNP MAS-NMR实验。在此开发项目的同时，我们将对冷冻RNA和蛋白质溶液、膜蛋白复合物和完整核糖体颗粒进行cwDNP MAS-NMR应用。通过对该研究项目实施开放用户概念，可以访问这个独特的研究基础设施。