DNP approach for structural determination of large protein-RNA complexes by solid-state NMR

点击翻译按钮获取中文摘要

In this research project we aim to investigate the feasibility of dynamic nuclear polarization (DNP) aided experiments for structural studies of large protein-RNA complexes by solid-state NMR. Solid-state NMR has been successfully applied for studies of amyloid fibrils, membrane proteins and large protein complexes; first ssNMR studies on protein-RNA complexes have been reported recently. While structural information on small protein-RNA complexes can be obtained in a reasonable time, studies of large complexes are much more time consuming - and often unfeasible - due to lack of sensitivity and severe signal overlap. DNP provides a significant boost in sensitivity for NMR experiments by transferring electron spin polarization of a paramagnetic polarizing agent to nuclear spins. NMR signal enhancement factors typically reach two orders of magnitude. This leads to time savings of the order of tens of thousands and opens the possibility to perform 3D and 4D experiments within few days. The unique collaboration of our research groups, being active in RNA structural biology by MAS NMR as well as DNP-enhanced MAS NMR, will allow us to develop methods for structural studies of large RNP complexes in close-to-native environment at atomic resolution. The methodology developed within this project will find broad applicability in several areas of structural biology.