Keivan Behjou Nasrin, Seyedalipour Bagher, Hosseini Faradonbeh Seyed Mahdi, Hosseinkhani Saman, Chaeichi Mohammad Javad
Department of Molecular and Cell Biology, Faculty of Basic Sciences, University of Mazandaran, Babolsar, Iran.
Department of Molecular and Cell Biology, Faculty of Basic Sciences, University of Mazandaran, Babolsar, Iran.
Bioorg Chem. 2025 Aug;163:108740. doi: 10.1016/j.bioorg.2025.108740. Epub 2025 Jul 12.
Ionic liquids (ILs) offer a diverse and tunable approach to inhibiting amyloid protein formation, providing new strategies to develop anti-amyloidogenic agents for amyloid-based diseases, as explored in protein-IL research. This study explores the formation of amyloid aggregates of the E49K mutant under amyloidogenic conditions and evaluates the inhibitory potential of ethylammonium bromide (EABr) as an anti-amyloidogenic agent relevant to ALS pathology. The effect of EABr was studied using molecular dynamics simulations, FTIR spectroscopy, ANS fluorescence, ThT fluorescence, and TEM imaging. EABr promotes the formation of compact structures by reducing the exposure of contagious hydrophobic pockets in the E49K mutant aggregates, as monitored by ANS fluorescence. EABr binds with moderate affinity to the E49K mutant forms, inhibiting fibrillation by stabilizing aggregation-prone regions, as shown in fluorescence quenching. The decrease in ThT fluorescence intensity and the inhibition of fibril formation in a concentration-dependent manner highlight the interaction of EABr with the E49K mutant throughout the incubation period. TEM images during the saturation phase provide compelling evidence that EABr inhibits the formation of amyloid fibrils in the E49K mutant, thus supporting ThT analysis results. These findings demonstrate that EABr can inhibit amyloid formation of the E49K SOD1 mutant in vitro, supporting its potential as a lead compound for further pharmacological studies.
离子液体（ILs）为抑制淀粉样蛋白形成提供了一种多样且可调节的方法，正如在蛋白质-离子液体研究中所探索的那样，为开发针对淀粉样蛋白相关疾病的抗淀粉样生成剂提供了新策略。本研究探讨了E49K突变体在淀粉样生成条件下淀粉样聚集体的形成，并评估了溴化乙铵（EABr）作为与肌萎缩侧索硬化症（ALS）病理相关的抗淀粉样生成剂的抑制潜力。使用分子动力学模拟、傅里叶变换红外光谱（FTIR）、ANS荧光、硫黄素T（ThT）荧光和透射电子显微镜（TEM）成像研究了EABr的作用。通过ANS荧光监测发现，EABr通过减少E49K突变体聚集体中传染性疏水口袋的暴露来促进紧密结构的形成。如荧光猝灭所示，EABr以中等亲和力与E49K突变体形式结合，通过稳定易聚集区域来抑制纤维化。ThT荧光强度的降低以及纤维形成的浓度依赖性抑制突出了EABr在整个孵育期与E49K突变体的相互作用。饱和阶段的TEM图像提供了令人信服的证据，表明EABr抑制了E49K突变体中淀粉样纤维的形成，从而支持了ThT分析结果。这些发现表明，EABr可以在体外抑制E49K超氧化物歧化酶1（SOD1）突变体的淀粉样形成，支持其作为进一步药理学研究先导化合物的潜力。
