Vernalis Research, Granta Park, Great Abington, Cambridge CB21 6GB, UK.
YSBL, Department of Chemistry, University of York, Heslington, York YO10 5DD, UK.
Int J Mol Sci. 2020 Jul 24;21(15):5257. doi: 10.3390/ijms21155257.
Over the past decade intrinsically disordered proteins (IDPs) have emerged as a biologically important class of proteins, many of which are of therapeutic relevance. Here, we investigated the interactions between a model IDP system, tau K18, and nine literature compounds that have been reported as having an effect on tau in order to identify a robust IDP-ligand system for the optimization of a range of biophysical methods. We used NMR, surface plasmon resonance (SPR) and microscale thermophoresis (MST) methods to investigate the binding of these compounds to tau K18; only one showed unambiguous interaction with tau K18. Several near neighbors of this compound were synthesized and their interactions with tau K18 characterized using additional NMR methods, including 1D ligand-observed NMR, diffusion-ordered spectroscopy (DOSY) and F NMR. This study demonstrates that it is possible to detect and characterize IDP-ligand interactions using biophysical methods. However, care must be taken to account for possible artefacts, particularly the impact of compound solubility and where the protein has to be immobilized.
在过去的十年中，无序蛋白质（IDPs）已成为一类具有重要生物学意义的蛋白质，其中许多与治疗相关。在这里，我们研究了模型 IDP 系统 tau K18 与九种文献化合物之间的相互作用，这些化合物已被报道对 tau 有影响，目的是为一系列生物物理方法的优化确定一个稳健的 IDP-配体系统。我们使用 NMR、表面等离子体共振（SPR）和微尺度热泳（MST）方法来研究这些化合物与 tau K18 的结合情况；只有一种化合物与 tau K18 表现出明确的相互作用。该化合物的几个近邻被合成，并使用其他 NMR 方法，包括 1D 配体观察 NMR、扩散有序光谱（DOSY）和 F NMR 来表征它们与 tau K18 的相互作用。这项研究表明，使用生物物理方法检测和表征 IDP-配体相互作用是可行的。然而，必须注意可能出现的假象，特别是化合物溶解度的影响以及蛋白质必须固定的位置。
