1H, 13C, and 15N resonance assignment of a 179 residue fragment of hepatitis C virus non-structural protein 5A
1H, 13C, and 15N resonance assignment of a 179 residue fragment of hepatitis C virus non-structural protein 5A
复制标题
丙型肝炎病毒非结构蛋白 5A 的 179 个残基片段的 1H、13C 和 15N 共振分配
DOI：
--
复制
发表时间：
2011
影响因子：
0.9
通讯作者：
B. Brutscher
中科院分区：
文献类型：
--
作者：
S. Feuerstein;Zsófia Sólyom;Amine Aladağ;S. Hoffmann;D. Willbold;B. Brutscher
非结构蛋白5A（NS 5A）在丙型肝炎病毒的生命周期中发挥着重要作用。这种富含脯氨酸的磷蛋白被组织成三个结构域。除了在病毒复制和病毒组装中的作用外，NS 5A还参与多种细胞调节过程。结构域2和3的最新研究表明，它们都属于本质无序的蛋白质类，因为它们采用天然的未折叠状态。特别是，结构域2及其邻近区域负责NS 5A与病毒持续存在所必需的其他蛋白质的多重相互作用。对于本质无序蛋白质观察到的低化学位移分散对核磁共振光谱提出了挑战。在这里，我们报告的NS 5A的179个残基的片段，包括整个结构域2的顺序共振分配，使用一组灵敏度和分辨率优化的3D相关实验，以及在1H-15 N相关光谱的氨基酸类型编辑。我们的分配揭示了几个片段具有形成α-螺旋结构的高倾向，这可能对该蛋白片段作为多功能相互作用平台的功能具有重要意义。
Non-structural protein 5A (NS5A) plays an important role in the life cycle of hepatitis C virus. This proline-rich phosphoprotein is organized into three domains. Besides its role in virus replication and virus assembly, NS5A is involved in a variety of cellular regulation processes. Recent studies on domain 2 and 3 revealed that both belong to the class of intrinsically disordered proteins as they adopt a natively unfolded state. In particular, domain 2 together with its vicinal regions is responsible for NS5A’s multiple interactions with other proteins necessary for virus persistence. The low chemical shift dispersion observed for instrinsically disordered proteins presents a challenge for NMR spectroscopy. Here we report sequential resonance assignment of a 179-residue fragment of NS5A, comprising the entire domain 2, using a set of sensitivity and resolution optimized 3D correlation experiments, as well as amino-acid-type editing in 1H-15N correlation spectra. Our assignment reveals the presence of several segments with high propensity to form α-helical structure that may be of importance to the function of this protein fragment as a versatile interaction platform.