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

通过固态 NMR 确定大蛋白质-RNA 复合物结构的 DNP 方法

• 批准号: 399211197
• 负责人: Professor Dr.-Ing. Björn Corzilius
• 金额: --
• 依托单位: Institut für Molekulare Biowissenschaften
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2018
• 资助国家: 德国
• 起止时间: 2017-12-31 至 2022-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/399211197?language=en
• 关键词: DNP; approach; structural; determination; large

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.

在该研究项目中，我们旨在研究动态核极化（DNP）的可行性（DNP）辅助实验，用于固态NMR对大蛋白RNA复合物的结构研究。固态NMR已成功地用于研究淀粉样蛋白原纤维，膜蛋白和大蛋白质复合物的研究。最近已经报道了有关蛋白-RNA复合物的首次SSNMR研究。尽管可以在合理的时间内获得有关小蛋白RNA复合物的结构信息，但由于缺乏敏感性和严重的信号重叠，对大型复合物的研究更加耗时（通常是不可行）。 DNP通过将顺磁性偏振剂的电子自旋极化转移到核自旋方面，从而显着提高了NMR实验的敏感性。 NMR信号增强因子通常达到两个数量级。这导致了成千上万的时间节省的时间，并在几天内开放了执行3D和4D实验的可能性。 MAS NMR和DNP增强的MAS NMR活跃于RNA结构生物学的独特合作将使我们能够开发用于原子分辨率接近本地环境中大型RNP复合物的结构研究的方法。该项目中开发的方法将在结构生物学的几个领域中找到广泛的适用性。