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

点击翻译按钮获取中文摘要

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.