Institute of Protein Research, Russian Academy of Sciences , Pushchino 142290, Russia.
State Research Center for Applied Microbiology & Biotechnology , Obolensk 142279, Russia.
Langmuir. 2018 Feb 13;34(6):2332-2343. doi: 10.1021/acs.langmuir.7b03393. Epub 2018 Jan 30.
To identify the key stages in the amyloid fibril formation we studied the aggregation of amyloidogenic fragments of Aβ peptide, Aβ(16-25), Aβ(31-40), and Aβ(33-42), using the methods of electron microscopy, X-ray analysis, mass spectrometry, and structural modeling. We have found that fragments Aβ(31-40) and Aβ(33-42) form amyloid fibrils in the shape of bundles and ribbons, while fragment Aβ(16-25) forms only nanofilms. We are the first who performed 2D reconstruction of amyloid fibrils by the Markham rotation technique on electron micrographs of negatively stained fragments of Aβ peptide. Combined analysis of the data allows us to speculate that both the fibrils and the films are formed via association of ring-shaped oligomers with the external diameter of about 6 to 7 nm, the internal diameter of 2 to 3 nm, and the height of ∼3 nm. We conclude that such oligomers are the main building blocks in fibrils of any morphology. The interaction of ring oligomers with each other in different ways makes it possible to explain their polymorphism. The new mechanism of polymerization of amyloidogenic proteins and peptides, described here, could stimulate new approaches in the development of future therapeutics for the treatment of amyloid-related diseases.
为了确定淀粉样纤维形成的关键阶段,我们使用电子显微镜、X 射线分析、质谱和结构建模等方法研究了淀粉样肽 Aβ 的淀粉样纤维形成片段 Aβ(16-25)、Aβ(31-40)和 Aβ(33-42)的聚集。我们发现片段 Aβ(31-40)和 Aβ(33-42)形成束状和带状的淀粉样纤维,而片段 Aβ(16-25)仅形成纳米薄膜。我们是第一个在负染色 Aβ 肽片段的电子显微镜照片上使用 Markham 旋转技术对淀粉样纤维进行二维重建的人。综合数据分析使我们推测,纤维和薄膜都是通过具有约 6 至 7nm 外径、2 至 3nm 内径和约 3nm 高度的环形寡聚物的缔合形成的。我们得出结论,这种寡聚物是任何形态纤维的主要构建块。环形寡聚物之间以不同的方式相互作用,使得它们的多态性成为可能。这里描述的淀粉样蛋白和肽的聚合的新机制,可以为开发治疗淀粉样相关疾病的未来治疗方法提供新的方法。