Institute of Biochemistry, Department of Biology, ETH Zurich, Hönggerbergring 64, 8093 Zürich, Switzerland.
Institute of Molecular Systems Biology, Department of Biology, ETH Zurich, Otto-Stern-Weg 3, 8093 Zürich, Switzerland.
Nucleic Acids Res. 2023 May 22;51(9):4555-4571. doi: 10.1093/nar/gkad195.
The pandemic caused by SARS-CoV-2 has called for concerted efforts to generate new insights into the biology of betacoronaviruses to inform drug screening and development. Here, we establish a workflow to determine the RNA recognition and druggability of the nucleocapsid N-protein of SARS-CoV-2, a highly abundant protein crucial for the viral life cycle. We use a synergistic method that combines NMR spectroscopy and protein-RNA cross-linking coupled to mass spectrometry to quickly determine the RNA binding of two RNA recognition domains of the N-protein. Finally, we explore the druggability of these domains by performing an NMR fragment screening. This workflow identified small molecule chemotypes that bind to RNA binding interfaces and that have promising properties for further fragment expansion and drug development.
由 SARS-CoV-2 引起的大流行要求我们共同努力，深入了解β冠状病毒的生物学特性，以为药物筛选和开发提供信息。在这里，我们建立了一个工作流程，以确定 SARS-CoV-2 的核衣壳 N 蛋白的 RNA 识别和可药性，该蛋白是病毒生命周期中至关重要的高度丰富蛋白。我们使用一种协同方法，将 NMR 光谱学与蛋白质-RNA 交联结合质谱联用，快速确定 N 蛋白的两个 RNA 识别结构域的 RNA 结合情况。最后，我们通过进行 NMR 片段筛选来探索这些结构域的可药性。该工作流程确定了与 RNA 结合界面结合的小分子化学型，它们具有进一步进行片段扩展和药物开发的有前途的特性。
