Institute of Multidisciplinary Research for Advanced Materials, Tohoku University, Katahira 2-1-1, Aoba-ku, Sendai, 980-8577, Japan.
Graduate School of Life Sciences, Tohoku University, Sendai, 980-8577, Japan.
Sci Rep. 2021 Mar 23;11(1):6643. doi: 10.1038/s41598-021-86098-1.
Liquid droplets of aggregation-prone proteins, which become hydrogels or form amyloid fibrils, are a potential target for drug discovery. In this study, we proposed an experiment-guided protocol for characterizing the design grammar of peptides that can regulate droplet formation and aggregation. The protocol essentially involves investigation of 19 amino acid additives and polymerization of the identified amino acids. As a proof of concept, we applied this protocol to fused in sarcoma (FUS). First, we evaluated 19 amino acid additives for an FUS solution and identified Arg and Tyr as suppressors of droplet formation. Molecular dynamics simulations suggested that the Arg additive interacts with specific residues of FUS, thereby inhibiting the cation-π and electrostatic interactions between the FUS molecules. Second, we observed that Arg polymers promote FUS droplet formation, unlike Arg monomers, by bridging the FUS molecules. Third, we found that the Arg additive suppressed solid aggregate formation of FUS, while Arg polymer enhanced it. Finally, we observed that amyloid-forming peptides induced the conversion of FUS droplets to solid aggregates of FUS. The developed protocol could be used for the primary design of peptides controlling liquid droplets and aggregates of proteins.
易于聚集的蛋白质的液滴会形成水凝胶或形成淀粉样纤维，这是药物发现的一个潜在目标。在这项研究中，我们提出了一个实验指导的方案，用于描述可以调节液滴形成和聚集的肽的设计语法。该方案主要涉及对 19 种氨基酸添加剂的研究和鉴定氨基酸的聚合。作为概念验证，我们将该方案应用于融合肉瘤蛋白（FUS）。首先，我们评估了 19 种 FUS 溶液中的氨基酸添加剂，并鉴定出 Arg 和 Tyr 是抑制液滴形成的抑制剂。分子动力学模拟表明，Arg 添加剂与 FUS 的特定残基相互作用，从而抑制 FUS 分子之间的阳离子-π 和静电相互作用。其次，我们观察到 Arg 聚合物通过桥接 FUS 分子促进 FUS 液滴的形成，而不是 Arg 单体。第三，我们发现 Arg 添加剂抑制了 FUS 的固体聚集体形成，而 Arg 聚合物增强了它。最后，我们观察到淀粉样形成肽诱导 FUS 液滴向 FUS 的固体聚集体转化。所开发的方案可用于控制蛋白质液滴和聚集体的肽的初步设计。
