Dipartimento di Scienze della Vita e dell'Ambiente , Università Politecnica delle Marche , Ancona 60122 , Italy.
Université Grenoble Alpes-Laboratoire TIMC/IMAG UMR CNRS 5525 , Grenoble 38000 , France.
J Phys Chem B. 2019 Jan 24;123(3):631-638. doi: 10.1021/acs.jpcb.8b11719. Epub 2019 Jan 9.
The understanding of amyloid β-peptide (Aβ) interactions with cellular membranes is a crucial molecular challenge against Alzheimer's disease. Indeed, Aβ prefibrillar oligomeric intermediates are believed to be the most toxic species, able to induce cellular damages directly by membrane damage. We present a neutron-scattering study on the interaction of large unilamellar vesicles (LUV), as cell membrane models, with both freshly dissolved Aβ and early toxic prefibrillar oligomers, intermediate states in the amyloid pathway. In addition, we explore the effect of coincubating the Aβ-peptide with the chaperonin Hsp60, which is known to strongly interact with it in its aggregation pattern. In fact, the interaction of the LUV with coincubated Aβ/Hsp60, right after mixing and after following the aggregation protocol leading to the toxic intermediates in the absence of Hsp60, is studied. Neutron spin echo experiments show that the interaction with both freshly dissolved and aggregate Aβ species brings about an increase in membrane stiffness, whereas the presence of even very low amounts of Hsp60 (ratio Aβ/Hsp60 = 25:1) maintains unaltered the elastic properties of the membrane bilayer. A coherent interpretation of these results, related to previous literature, can be based on the ability of the chaperonin to interfere with Aβ aggregation, by the specific recognition of the Aβ-reactive transient species. In this framework, our results strongly suggest that early in a freshly dissolved Aβ solution are present some species able to modify the bilayer dynamics, and the chaperonin plays the role of an assistant in such stochastic "misfolding events", avoiding the insult on the membrane as well as the onset of the aggregation cascade.
对淀粉样β肽(Aβ)与细胞膜相互作用的理解是对抗阿尔茨海默病的一个关键分子挑战。事实上,Aβ原纤维前聚集态中间体被认为是最具毒性的物种,能够通过膜损伤直接诱导细胞损伤。我们进行了一项中子散射研究,研究了大单层囊泡(LUV)与新鲜溶解的 Aβ和早期毒性原纤维前聚集态中间体(淀粉样途径中的中间状态)之间的相互作用。此外,我们还探索了将 Aβ 肽与伴侣蛋白 Hsp60 共孵育的效果,已知 Hsp60 在其聚集模式中与 Aβ 强烈相互作用。事实上,研究了 LUV 与共孵育的 Aβ/Hsp60 的相互作用,在混合后以及在没有 Hsp60 的情况下遵循导致毒性中间体的聚集方案之后。中子自旋回波实验表明,与新鲜溶解的和聚集的 Aβ 物种的相互作用都会导致膜刚性增加,而即使存在非常低量的 Hsp60(Aβ/Hsp60 比为 25:1)也能保持膜双层的弹性特性不变。这些结果与之前的文献相关,可以基于伴侣蛋白通过对 Aβ 反应性瞬态物种的特异性识别来干扰 Aβ 聚集的能力进行一致解释。在这种情况下,我们的结果强烈表明,在新鲜溶解的 Aβ 溶液中,存在一些能够改变双层动力学的物种,而伴侣蛋白在这种随机的“错误折叠事件”中起到辅助作用,从而避免了对膜的损伤以及聚集级联的发生。