The Disordered Region of the HCV Protein NS5A: Conformational Dynamics, SH3 Binding, and Phosphorylation.
The Disordered Region of the HCV Protein NS5A: Conformational Dynamics, SH3 Binding, and Phosphorylation.
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HCV 蛋白 NS5A 的无序区域：构象动力学、SH3 结合和磷酸化
DOI：
10.1016/j.bpj.2015.06.040
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发表时间：
2015
影响因子：
3.4
通讯作者：
Brutscher B
中科院分区：
文献类型：
--
作者：
Solyom Z;Schwarten M;Bosco M;Polidori A;Durand G;Willbold D;Brutscher B
内在无序蛋白（IDPs）在没有明确定义的三维结构的情况下发挥其生理作用。然而，残馀结构和构象动力学对其功能机制至关重要。例如，瞬态二级结构区域通常参与分子识别，结合后结构稳定（或不稳定）。另一方面，远程相互作用决定了IDP的流体动力学半径，从而决定了蛋白质通过所谓的飞投机制捕获结合伙伴的距离。远程相互作用的调节也提供了一种方便的方法来微调蛋白质的相互作用网络，通过使结合位点或多或少地可接近。本研究主要利用核磁共振波谱技术研究了丙型肝炎病毒（HCV）非结构蛋白5A （NS5A）的残馀二级结构和远程相互作用，这是一种典型的病毒IDP，在病毒生命周期中具有多种功能。NS5A包括一个n端折叠结构域，随后是一个大的（约250个残基）无序的c端部分。将全长NS5A的核磁共振波谱与仅由c端残基191-447 （NS5A- d2d3）组成的蛋白结构体的核磁共振波谱进行比较，我们可以得出结论，NS5A的球状部分和无序部分之间没有明显的相互作用。NS5A-D2D3虽然整体柔韧性较高，但存在较大程度的局部残留（α-螺旋和β-旋）结构，以及静电远程相互作用网络。此外，我们可以证明这些远程相互作用在与宿主蛋白Bin1结合以及NS5A被CK2磷酸化后被调节。由于参与这些相互作用的带电肽区在不同的HCV基因型中是保守的，这些短暂的远程相互作用可能对NS5A在HCV生命周期中的一些功能很重要。
Intrinsically disordered proteins (IDPs) perform their physiological role without possessing a well-defined three-dimensional structure. Still, residual structure and conformational dynamics of IDPs are crucial for the mechanisms underlying their functions. For example, regions of transient secondary structure are often involved in molecular recognition, with the structure being stabilized (or not) upon binding. Long-range interactions, on the other hand, determine the hydrodynamic radius of the IDP, and thus the distance over which the protein can catch binding partners via so-called fly-casting mechanisms. The modulation of long-range interactions also presents a convenient way of fine-tuning the protein's interaction network, by making binding sites more or less accessible. Here we studied, mainly by nuclear magnetic resonance spectroscopy, residual secondary structure and long-range interactions in nonstructural protein 5A (NS5A) from hepatitis C virus (HCV), a typical viral IDP with multiple functions during the viral life cycle. NS5A comprises an N-terminal folded domain, followed by a large (∼250-residue) disordered C-terminal part. Comparing nuclear magnetic resonance spectra of full-length NS5A with those of a protein construct composed of only the C-terminal residues 191–447 (NS5A-D2D3) allowed us to conclude that there is no significant interaction between the globular and disordered parts of NS5A. NS5A-D2D3, despite its overall high flexibility, shows a large extent of local residual (α-helical andβ-turn) structure, as well as a network of electrostatic long-range interactions. Furthermore, we could demonstrate that these long-range interactions become modulated upon binding to the host protein Bin1, as well as after NS5A phosphorylation by CK2. As the charged peptide regions involved in these interactions are well conserved among the different HCV genotypes, these transient long-range interactions may be important for some of the functions of NS5A over the course of the HCV life cycle.
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HCV NS5A 蛋白的结构域 2 本质上是非结构化的。
影响因子：
1.6
作者：
X. Hanoulle;Aurélie Badillo;Dries Verdegem;F. Penin;G. Lippens
通讯作者：
G. Lippens
影响因子：
2.1
作者：
Aladag, Amine;Hoffmann, Silke;Schwarten, Melanie
通讯作者：
Schwarten, Melanie
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
影响因子：
0.9
作者：
S. Feuerstein;Zsófia Sólyom;Amine Aladağ;S. Hoffmann;D. Willbold;B. Brutscher
通讯作者：
B. Brutscher
影响因子：
5.6
作者：
Feuerstein, Sophie;Solyom, Zsofia;Brutscher, Bernhard
通讯作者：
Brutscher, Bernhard
影响因子：
15
作者：
Lescop, Ewen;Rasia, Rodolfo;Brutscher, Bernhard
通讯作者：
Brutscher, Bernhard