Magnetic Resonance Center (CERM), University of Florence, Via Luigi Sacconi 6, 50019, Sesto Fiorentino, Italy.
Department of Chemistry "Ugo Schiff", University of Florence, Via della Lastruccia 3-13, 50019, Sesto Fiorentino, Italy.
Sci Rep. 2020 Nov 11;10(1):19574. doi: 10.1038/s41598-020-76522-3.
Using SAXS and NMR spectroscopy, we herein provide a high-resolution description of the intrinsically disordered N-terminal domain (PNT, aa 1-406) shared by the Nipah virus (NiV) phosphoprotein (P) and V protein, two key players in viral genome replication and in evasion of the host innate immune response, respectively. The use of multidimensional NMR spectroscopy allowed us to assign as much as 91% of the residues of this intrinsically disordered domain whose size constitutes a technical challenge for NMR studies. Chemical shifts and nuclear relaxation measurements provide the picture of a highly flexible protein. The combination of SAXS and NMR information enabled the description of the conformational ensemble of the protein in solution. The present results, beyond providing an overall description of the conformational behavior of this intrinsically disordered region, also constitute an asset for obtaining atomistic information in future interaction studies with viral and/or cellular partners. The present study can thus be regarded as the starting point towards the design of inhibitors that by targeting crucial protein-protein interactions involving PNT might be instrumental to combat this deadly virus.
通过使用小角 X 射线散射（SAXS）和核磁共振（NMR）光谱技术，我们在此提供了关于在尼帕病毒（NiV）磷蛋白（P）和 V 蛋白之间共享的固有无序 N 端结构域（PNT，aa 1-406）的高分辨率描述。这两种蛋白分别是病毒基因组复制和逃避宿主先天免疫反应的关键因素。多维 NMR 光谱技术的使用使我们能够对这个固有无序结构域的 91%的残基进行分配，而该结构域的大小对 NMR 研究来说是一个技术挑战。化学位移和核弛豫测量提供了一种高度灵活的蛋白质图像。SAXS 和 NMR 信息的结合能够描述该蛋白质在溶液中的构象集合。目前的结果不仅提供了对这个固有无序区域构象行为的总体描述，而且还为未来与病毒和/或细胞伙伴的相互作用研究中获得原子信息提供了帮助。因此，本研究可以被视为设计抑制剂的起点，通过靶向涉及 PNT 的关键蛋白-蛋白相互作用，这些抑制剂可能有助于对抗这种致命病毒。
