Goud Alladi Charanraj, Kozlov Ihor, Skoupilová Patricie, Malina Lukáš, Roy Sudeep, Das Viswanath
Institute of Molecular and Translational Medicine, Faculty of Medicine and Dentistry, Palacký University and University Hospital Olomouc, Olomouc, Czech Republic.
Department of Medical Biophysics, Faculty of Medicine and Dentistry, Palacký University Olomouc, Olomouc, Czech Republic.
Protein Sci. 2025 Sep;34(9):e70240. doi: 10.1002/pro.70240.
Tau aggregation driven by microtubule-associated protein tau (MAPT) mutations is central to frontotemporal dementia pathology, yet no disease-modifying therapies effectively target mutant tau. Here, we identify purpurin (PUR) and oleocanthal (OLC) as selective inhibitors of mutant tau aggregation using peptide models spanning the R2R3 interface. Biophysical and cellular assays demonstrated that both compounds more effectively inhibit the aggregation of mutant tau peptides compared to wild-type, with PUR preferentially targeting V287I and N279K variants, and OLC showing broader inhibitory activity. Surface plasmon resonance and docking analyses revealed more stable interactions and lower binding free energies with mutant tau, consistent with their enhanced inhibitory effects. Computational studies using monomeric and fibrillar tau structures supported the mutation-specific binding profiles of PUR and OLC. Atomic force microscopy and confocal imaging confirmed reduced fibril formation, while post-transduction treatment assays showed that both compounds significantly suppressed intracellular tau propagation. Additionally, OLC reduced tau phosphorylation and oligomerization in SY5Y-TauP301L-EGFP cells expressing mutant tau. These findings highlight the potential of PUR and OLC as structurally distinct, mutation-targeted inhibitors of tau aggregation and propagation, providing a rationale for their further development as candidate therapeutics for frontotemporal dementia.
由微管相关蛋白tau(MAPT)突变驱动的tau蛋白聚集是额颞叶痴呆病理学的核心,但尚无有效的疾病修饰疗法能够有效靶向突变型tau蛋白。在此,我们使用跨越R2R3界面的肽模型,鉴定出紫红素(PUR)和油橄榄苦素(OLC)为突变型tau蛋白聚集的选择性抑制剂。生物物理和细胞实验表明,与野生型相比,这两种化合物都能更有效地抑制突变型tau肽的聚集,PUR优先靶向V287I和N279K变体,而OLC表现出更广泛的抑制活性。表面等离子体共振和对接分析揭示了与突变型tau蛋白更稳定的相互作用和更低的结合自由能,这与它们增强的抑制作用一致。使用单体和纤维状tau蛋白结构的计算研究支持了PUR和OLC的突变特异性结合谱。原子力显微镜和共聚焦成像证实了纤维形成减少,而转导后处理实验表明,这两种化合物都能显著抑制细胞内tau蛋白的传播。此外,OLC降低了表达突变型tau蛋白的SY5Y-TauP301L-EGFP细胞中tau蛋白的磷酸化和寡聚化。这些发现突出了PUR和OLC作为结构上不同的、针对突变的tau蛋白聚集和传播抑制剂的潜力,为它们作为额颞叶痴呆候选治疗药物的进一步开发提供了理论依据。