Department of Chemistry, University of Rochester, Rochester, New York 14627, USA.
ACS Chem Neurosci. 2012 Mar 21;3(3):211-20. doi: 10.1021/cn2001188. Epub 2012 Jan 9.
Amyloid-β (Aβ) self-assembly into cross-β amyloid fibrils is implicated in a causative role in Alzheimer's disease pathology. Uncertainties persist regarding the mechanisms of amyloid self-assembly and the role of metastable prefibrillar aggregates. Aβ fibrils feature a sheet-turn-sheet motif in the constituent β-strands; as such, turn nucleation has been proposed as a rate-limiting step in the self-assembly pathway. Herein, we report the use of an azobenzene β-hairpin mimetic to study the role turn nucleation plays on Aβ self-assembly. [3-(3-Aminomethyl)phenylazo]phenylacetic acid (AMPP) was incorporated into the putative turn region of Aβ42 to elicit temporal control over Aβ42 turn nucleation; it was hypothesized that self-assembly would be favored in the cis-AMPP conformation if β-hairpin formation occurs during Aβ self-assembly and that the trans-AMPP conformer would display attenuated fibrillization propensity. It was unexpectedly observed that the trans-AMPP Aβ42 conformer forms fibrillar constructs that are similar in almost all characteristics, including cytotoxicity, to wild-type Aβ42. Conversely, the cis-AMPP Aβ42 congeners formed nonfibrillar, amorphous aggregates that exhibited no cytotoxicity. Additionally, cis-trans photoisomerization resulted in rapid formation of native-like amyloid fibrils and trans-cis conversion in the fibril state reduced the population of native-like fibrils. Thus, temporal photocontrol over Aβ turn conformation provides significant insight into Aβ self-assembly. Specifically, Aβ mutants that adopt stable β-turns form aggregate structures that are unable to enter folding pathways leading to cross-β fibrils and cytotoxic prefibrillar intermediates.
淀粉样蛋白-β(Aβ)自组装成交叉-β淀粉样纤维与阿尔茨海默病病理学中的致病作用有关。关于淀粉样蛋白自组装的机制和亚稳前纤维状聚集体的作用仍然存在不确定性。Aβ纤维在组成的β-链中具有片层-转折-片层基序;因此,转折成核被提出是自组装途径中的限速步骤。在此,我们报告了使用偶氮苯β发夹类似物来研究转折成核在 Aβ自组装中的作用。[3-(3-氨甲基)苯基偶氮]苯乙酸(AMPP)被掺入 Aβ42 的假定转折区域,以引起对 Aβ42 转折成核的时间控制;假设如果β发夹形成发生在 Aβ自组装过程中,那么 Aβ42 顺式-AMPP 构象将有利于自组装,而反式-AMPP 构象将显示出减弱的纤维化倾向。出乎意料的是,观察到反式-AMPP Aβ42 构象形成的纤维状结构在几乎所有特征上都与野生型 Aβ42 相似,包括细胞毒性。相反,顺式-AMPP Aβ42 同系物形成无纤维状、无定形聚集体,没有细胞毒性。此外,顺-反异构化导致天然样淀粉样纤维的快速形成,而在纤维状态下的反-顺转换降低了天然样纤维的数量。因此,对 Aβ 转折构象的时间光控提供了对 Aβ 自组装的重要见解。具体而言,采用稳定β-转折的 Aβ 突变体形成无法进入导致交叉-β纤维和细胞毒性前纤维状中间产物的折叠途径的聚集体结构。