Department of Physical Chemistry, Faculty of Science, University of Santiago de Compostela, Lugo, Spain.
Methods Mol Biol. 2023;2551:1-14. doi: 10.1007/978-1-0716-2597-2_1.
Alzheimer's disease (AD) is a progressive neurodegenerative disease affecting cognitive and memory abilities and is believed to be linked to the formation and accumulation of neurotoxic aggregates of the Amyloid-β peptide (Aβ). In particular, it is the formation of soluble pre-fibrillar oligomers within the early stage of Aβ aggregation which is thought to represent a key step in the development of AD, thus underlining the interest in characterizing the aggregation process and the nature of these aggregates. In this context, fluorescence correlation spectroscopy (FCS) has emerged as a valuable alternative for the study of these systems in solution. Indeed, the use of FCS to study terminally labelled Aβ provides a means to detect changes in the size and concentration of initially monomeric Aβ samples by monitoring these fluorescently labelled species freely diffusing in solution with single-molecule resolution. Herein, we show how to employ FCS to study the early aggregation process of Aβ(1-42) and how this can be used to estimate the critical concentration for oligomer formation and to characterize the aggregates formed.
阿尔茨海默病(AD)是一种进行性神经退行性疾病,影响认知和记忆能力,据信与神经毒性淀粉样蛋白-β肽(Aβ)聚集体的形成和积累有关。特别是,可溶性预纤维寡聚物在 Aβ 聚集的早期形成被认为是 AD 发展的关键步骤,因此强调了对聚集过程和这些聚集体性质进行表征的兴趣。在这种情况下,荧光相关光谱(FCS)已成为研究溶液中这些系统的一种有价值的替代方法。实际上,使用 FCS 研究末端标记的 Aβ 提供了一种通过监测在溶液中自由扩散的这些荧光标记的物种来检测初始单体 Aβ 样品的大小和浓度变化的方法,具有单分子分辨率。本文展示了如何使用 FCS 研究 Aβ(1-42)的早期聚集过程,以及如何利用它来估计寡聚物形成的临界浓度并表征形成的聚集体。