Small-molecule binding to an intrinsically disordered protein revealed by experimental NMR 19 F transverse spin-relaxation
Small-molecule binding to an intrinsically disordered protein revealed by experimental NMR 19 F transverse spin-relaxation
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实验 NMR 19 F 横向自旋弛豫揭示了小分子与本质上无序的蛋白质的结合
DOI：
10.1101/2023.05.03.539297
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Heller G
中科院分区：
文献类型：
--
作者：
Heller G
内位无序蛋白质是高度动态的生物分子，其在许多结构构象之间快速相互转换。传统上，这些蛋白质被认为是不可药用的，因为它们缺乏经典的长寿命结合口袋。最近的证据表明，本质上无序的蛋白质可以结合小的药物样分子，然而，有有限的方法来表征这些相互作用的实验。在这里，我们表明，配体检测19 F横向弛豫速率（R2）从核磁共振光谱得到的小分子和一个内在无序的蛋白质之间的相互作用是高度敏感的，相反，化学位移扰动是最低限度的敏感，这种相互作用。用这种方法，我们发现5-氟吲哚小分子与丙型肝炎病毒非结构蛋白5A的无序结构域相互作用，其作用的Kd值为260 ± 110 μM。我们还表明，5-氟吲哚保持高度动态的结合形式。我们的研究结果表明，ligand-detected 19 F横向弛豫测量可以代表一个非常有效的筛选策略，以确定能够与这些传统上难以捉摸的，动态的生物分子相互作用的分子。
Intrinsically disordered proteins are highly dynamic biomolecules that rapidly interconvert between many structural conformations. Traditionally, these proteins have been considered un-druggable because of their lack of classical long-lived binding pockets. Recent evidence suggests that intrinsically disordered proteins can bind small, drug-like molecules, however, there are limited approaches to characterize these interactions experimentally. Here we demonstrate that ligand-detected19F transverse relaxation rates (R2) obtained from Nuclear Magnetic Resonance spectroscopy are highly sensitive to the interaction between a small-molecule and an intrinsically disordered protein, in contrast to chemical shift perturbations which are minimally sensitive for this interaction. With this method, we show that the small molecule, 5-fluoroindole, interacts with the disordered domains of non-structural protein 5A from hepatitis C virus with aKdof 260 ± 110 μM. We also demonstrate that 5-fluoroindole remains highly dynamic in the bound form. Our findings suggest that ligand-detected19F transverse relaxation measurements could represent a highly effective screening strategy to identify molecules capable of interacting with these traditionally elusive, dynamic biomolecules.