蛋白质的淀粉样纤维化与哺乳动物神经退行性疾病的发生密切相关，研究表明蛋白在淀粉样纤维化早期所形成的寡聚体是致病组分，而淀粉样蛋白的寡聚化机制目前尚无定论。由于寡聚体具有不稳定、瞬态性、不均一性等特点，使得系综水平的技术手段在淀粉样蛋白寡聚化机制的研究中有局限性。本项目拟以酵母Ure2作为淀粉样蛋白研究模型，应用单分子荧光共振能量传递，研究Ure2寡聚化动力学，以及寡聚体组装形式变化；应用荧光共振能量传递结合荧光相关光谱技术，研究Ure2寡聚体的大小及其在纤维化过程中的变化；应用单分子荧光共振能量传递结合单分子荧光偏振，研究Ure2在寡聚化过程中，分子内部的构象变化机制及结构域之间相互作用。本项目的研究成果将有助于深入理解淀粉样蛋白的寡聚机制，为开发治疗神经退行性疾病的药物提供新线索。

The phenomenon of amyloid fibril formation is closely related to mammalian neurodegenerative disease. It has been reported that the protein oligomers formed early during the process of amyloid fibril formation are the pathogenic species. However, the mechanism of the amyloid protein oligomerization process is still an open question. Study of the mechanism of oligomerization using traditional ensemble techniques is challenging, due to the metastable, transient, and heterogenous nature of the oligomers. In this project, using the yeast prion protein Ure2 as an amyloid model, we plan to study the kinetics of Ure2 oligomerization and structural reorganization within the oligomers using single molecule FRET. We also plan to apply the FRET-FCS technique to analyse the size of Ure2 oligomers and how this changes during the fibril formation process. Finally, we will investigate the conformational change and interdomain interaction within the Ure2 monomers during oligomerization by using a combination of single molecule FRET and single molecule polarization. This research will contribute to understanding of the mechanism of protein oligomerization, and will provide new clues to drug development for neurodegenerative disease.