Departamento de Química Física, Instituto de Biotecnología y Unidad de Excelencia de Química Aplicada a Biomedicina y Medioambiente (UEQ), Facultad de Ciencias, Universidad de Granada, Granada, Spain.
Methods Mol Biol. 2023;2551:15-28. doi: 10.1007/978-1-0716-2597-2_2.
Amyloid aggregation is a hallmark in many neuropathologies and other diseases of tremendous impact. It is increasingly evident that neuronal death associated with Alzheimer's disease (AD) is mainly produced by oligomers of the amyloid-β (Aβ) peptide. Yet little is known about the detailed structural and biophysical mechanisms of their formation. This lack of complete understanding comes from the labile nature and handling complexity of the oligomers. Consequently, providing reproducible and robust protocols for oligomer preparation is of particular importance.In this study, we describe detailed methods for the preparation and isolation of micellar oligomers of Aβ that evolve towards larger and more stable oligomers enriched in beta-sheet structure and able to acquire a higher capacity to fibrillate. We also describe briefly some biophysical experiments allowing oligomer characterization.
淀粉样蛋白聚集是许多神经病理学和其他具有巨大影响的疾病的标志。越来越明显的是，与阿尔茨海默病（AD）相关的神经元死亡主要是由淀粉样β（Aβ）肽的低聚物产生的。然而，关于它们形成的详细结构和生物物理机制知之甚少。这种不完全理解来自于低聚物的不稳定性质和处理复杂性。因此，提供可重复和稳健的低聚物制备方案尤为重要。在这项研究中，我们描述了制备和分离 Aβ胶束低聚物的详细方法，这些低聚物会向富含β-折叠结构且更稳定的、具有更高纤维形成能力的大而稳定的低聚物进化。我们还简要描述了一些允许低聚物表征的生物物理实验。
