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

• Subject Area: Basic Research in Biology and Medicine
• Term: Funded in 2022
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 497967936

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.

• DFG Programme: Major Research Instrumentation
• Major Instrumentation: 600 MHz NMR-Spektrometer-Upgrade plus Kryo-Plattform und Kryo-Probenkopf (Teilfinanzierung)
• Instrumentation Group: 1740 Hochauflösende NMR-Spektrometer
• Applicant Institution: Heinrich-Heine-Universität Düsseldorf
• Leader: Professor Dieter Willbold, Ph.D.