真核生物细胞囊泡运输过程中的膜融合依赖SNARE复合体的解聚再利用循环，但其解聚的受力位点及作用机制尚不清楚。我们前期研究成功解析了20S超复合体中α-SNAP-SNARE亚组装体3.9 Å与NSF-D1D2结构域3.7 Å的高分辨三维结构，确定了参与解聚的α-SNAP蛋白施力位点，以及SNARE螺旋束N端与NSF-D1结构域中心孔之间的相互作用。为此我们推测SNARE复合体在解聚过程中受到以其N端作用为支点与抓手的解旋与剪切。本项目依据课题组前期结构模型，结合人群神经退行性疾病突变数据，构建不同的SNARE蛋白定点与N端截短突变体，利用电生理、冷冻电镜、化学交联质谱等手段，探究突变对SNARE复合体解聚及20S超复合体结构的影响，阐明SNARE复合体在解聚过程中的受力模式，对揭示SNARE突变通过影响解聚循环效率引发疾病的结构基础具有重要意义，并为相关的药物设计与开发提供线索和理论依据。

Membrane fusion during the vesicle trafficking of eukaryotes depends on the disassembly to recycle of SNARE complex, but the stress sites and the mechanism of disassembly remain still unclear. Our previous work has figured out the high-resolution three-dimensional structure of α-SNAP-SNARE subcomplex at 3.9 Å and NSF-D1D2 domain at 3.7 Å in 20S super complex by cryo-electron microscope (Cryo-EM), which helps us confirm the drive site of α-SNAP involved in SNARE disassembly and the interplay between the N-terminus of SNARE helical bundle and the central pore of NSF-D1 domain. Accordingly, we speculate that SNARE complex is subjected to unwinding and shearing in the process of disassembly with its N-terminal interaction as the fulcrum and gripper. Based on the built atomic model and combined with population data on neurodegenerative disease mutations, this study will construct a series of site-directed and N-terminus truncation SNARE mutants to explore the effects of mutations on the disassembly of SNARE complex and the structure of 20S super complex, by means of electrophysiology, Cryo-EM and chemical cross-linking mass spectrometry. To elucidate the stress pattern of SNARE complex in the disassembly is of great significance to reveal the structural basis of SNARE mutation causing diseases by hindering the disassembly cycle, and will provide clues and theoretical basis for relevant drug design and development.