Realignment of a 600 MHz NMR system for cellular applications and antibiotic development

重新调整 600 MHz NMR 系统，用于细胞应用和抗生素开发

• 批准号: 497967936
• 金额: --
• 依托单位: Heinrich-Heine-Universität Düsseldorf
• 依托单位国家: 德国
• 项目类别: Major Research Instrumentation
• 财政年份: 2022
• 资助国家: 德国
• 起止时间: 2021-12-31 至 无数据
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/497967936?language=en
• 关键词: Realignment; 600; MHz; NMR; system

The main goal of the project is to make the outstanding properties of fluorine atoms (19F) available for NMR-based structural investigations for the research groups at HHU. The measure therefore represents an essential application extension of the Biomolecular NMR Center, which has been in existence since 2004, and will enable high-resolution insights into cellular systems as well as significantly more efficient antibiotic development. Currently, there is no adequate possibility at HHU to use 19F nuclear spins in biological macromolecules and in living systems. However, a steadily growing body of research from other NMR centers confirms the broad applicability and tremendous scientific utility of 19F nuclear spin. The real benefit of the 19F detection technique stems from the fact that fluorine atoms, on the one hand, have very good NMR properties (100% occurrence of the NMR active isotope, high signal strength and strong dependence of the NMR frequency on the chemical environment). Furthermore, 19F is present only in negligible amounts in biological systems, such as eukaryotic cells. However, there are numerous ways to specifically label molecules with 19F. This allows, for example, highly sensitive and high-resolution NMR studies of 19F-labeled molecules of their native cellular environment, since virtually no background signal emanates from the cell. This is not the case for the regular NMR active nuclei of biological systems (1H, 13C, 15N, 31P). Therefore, concrete planned 19F-NMR applications include the investigation of the structure and dynamics of neuropeptides, G-protein coupled receptors, and DNA-mediated catalysis using so-called in-cell NMR methods. In addition, in-cell NMR will be further developed by using not only isolated cells but also, for the first time, so-called three-dimensional organoids, such as brain organoids.In addition to in-cell NMR, 19F NMR spectroscopy also offers unique possibilities for in vitro investigations, such as the characterization of nucleic acid structures or the screening of protein-ligand interactions. For the latter, an automated sample changer is also to be acquired as part of the measure, which should enable efficient screening of substance (fragment) e.g. for the development of a new class of antibiotics. The new in-cell NMR capabilities can be ideally combined with cryo-electron tomography. The planned combination of in-cell NMR and cryo-electron tomography will generate an outstanding added value for both foci of HHU, which is only available at a few locations worldwide.This measure is a new procurement that will serve to convert an already existing and fully intact 14.1 Tesla superconducting 600 MHz NMR magnet into a contemporary tool for in-cell and in-organoid NMR that is useful for the site.

该项目的主要目标是使 HHU 研究小组能够利用氟原子 (19F) 的出色特性进行基于 NMR 的结构研究。因此，该措施代表了自 2004 年成立的生物分子 NMR 中心的重要应用扩展，并将实现对细胞系统的高分辨率洞察以及显着更有效的抗生素开发。目前，HHU 还没有足够的可能性在生物大分子和生命系统中使用 19F 核自旋。然而，来自其他 NMR 中心的稳定增长的研究证实了 19F 核自旋的广泛适用性和巨大的科学实用性。 19F检测技术的真正好处源于这样一个事实：氟原子一方面具有非常好的NMR特性（NMR活性同位素100%出现、高信号强度以及NMR频率对化学环境的强烈依赖性） ）。此外，19F在生物系统（例如真核细胞）中的存在量可忽略不计。然而，有多种方法可以用 19F 特异性标记分子。例如，这允许对其天然细胞环境中的 19F 标记分子进行高灵敏度和高分辨率的 NMR 研究，因为实际上细胞中没有发出背景信号。对于生物系统的常规 NMR 活性核（1H、13C、15N、31P）而言，情况并非如此。因此，具体计划的 19F-NMR 应用包括使用所谓的细胞内 NMR 方法研究神经肽、G 蛋白偶联受体和 DNA 介导的催化的结构和动力学。此外，细胞内核磁共振将得到进一步发展，不仅使用分离的细胞，还首次使用所谓的三维类器官，例如脑类器官。除了细胞内核磁共振之外，19F核磁共振波谱也将得到发展。为体外研究提供了独特的可能性，例如核酸结构的表征或蛋白质-配体相互作用的筛选。对于后者，还需要购买自动样品更换器作为测量的一部分，这应该能够有效筛选物质（片段），例如。用于开发一类新抗生素。新的细胞内核磁共振功能可以与冷冻电子断层扫描完美结合。计划将细胞核磁共振和冷冻电子断层扫描相结合，将为 HHU 的两个焦点产生显着的附加值，而 HHU 仅在全球少数几个地点提供。这项措施是一项新​​的采购，将有助于转换现有的和完好无损的 14.1 Tesla 超导 600 MHz NMR 磁体成为现代细胞内和类器官 NMR 工具，对现场有用。