Department of Biotechnology, University of Verona, 37134 Verona, Italy.
ACZON srl, Monte San Pietro, BO 40050, Italy.
Bioconjug Chem. 2022 Jul 20;33(7):1261-1268. doi: 10.1021/acs.bioconjchem.2c00168. Epub 2022 Jun 10.
Intrinsically disordered proteins (IDPs) are increasingly found to be associated with irreversible neurodegenerative disorders. The protein tau is a prototypical IDP whose abnormal aggregation into insoluble filaments is a major hallmark of Alzheimer's disease. The view has emerged that aggregation may proceed via alternative pathways involving oligomeric intermediates or phase-separated liquid droplets. Nanoparticles (NPs) offer significant potential for probing the mechanisms of protein fibrillation and may be capable of redirecting conformational transitions. Here, we camouflaged dye-doped silica NPs through functionalization with tau molecules to impart them the ability to associate with protein assemblies such as aggregates or condensates. The prepared NP-tau conjugates showed little influence on the aggregation kinetics and morphology of filamentous aggregates of tau but were found to associate with the filaments. Moreover, NP-tau conjugates were recruited and concentrated into polyanion-induced condensates of tau, driven by multivalent electrostatic interactions, thereby illuminating liquid droplets and their time-dependent transformation, as observed by fluorescence microscopy. NP-tau conjugates were capable of entering human neuroglioma cells and were not cytotoxic. Hence, we propose that NP-tau conjugates could serve as nanotracers for in vitro and in-cell studies to target and visualize tau assemblies and condensates, contributing to an explanation for the molecular mechanisms of abnormal protein aggregation.
内在无序蛋白质(IDPs)越来越多地与不可逆的神经退行性疾病有关。tau 蛋白是一种典型的 IDP,其异常聚集形成不溶性纤维是阿尔茨海默病的主要标志。目前的观点认为,聚集可能通过涉及寡聚中间体或相分离液滴的替代途径进行。纳米颗粒(NPs)为探测蛋白质纤化机制提供了重要的潜力,并且可能能够重新引导构象转变。在这里,我们通过用 tau 分子功能化来伪装染料掺杂的二氧化硅 NPs,赋予它们与蛋白质聚集体(如纤维状聚集体或凝聚物)结合的能力。制备的 NP-tau 缀合物对 tau 的丝状聚集体的聚集动力学和形态几乎没有影响,但被发现与纤维结合。此外,由于多价静电相互作用,NP-tau 缀合物被招募并浓缩到多阴离子诱导的 tau 凝聚物中,从而通过荧光显微镜观察到液滴及其随时间的转化。NP-tau 缀合物能够进入人神经胶质瘤细胞,并且没有细胞毒性。因此,我们提出 NP-tau 缀合物可以作为纳米示踪剂用于体外和细胞内研究,以靶向和可视化 tau 聚集体和凝聚物,有助于解释异常蛋白质聚集的分子机制。