Structural analysis of SAA-amyloid fibrils by cryo-EM and NMR

通过冷冻电镜和核磁共振对 SAA 淀粉样原纤维进行结构分析

• 批准号: 313913757
• 负责人: Professor Dr. Bernd Reif
• 金额: --
• 依托单位: Professur für Festkörper-NMR-Spektroskopie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2016
• 资助国家: 德国
• 起止时间: 2015-12-31 至 2021-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/313913757?language=en
• 关键词: Structural; analysis; SAA; amyloid; fibrils

AA amyloidosis presents, among all protein misfolding diseases, unique advantages for studying the structure, formation and propagation of pathogenic amyloids. There are highly relevant in vitro, cell culture and mouse model systems to reproduce crucial steps of amyloid fibril formation. Amyloid deposits are comparatively large-sized and amenable for biochemical analysis. Murine SAA1 is an extracellular protein and small enough to be well accessible for NMR. These features enable us to take an integrative structural biology approach in which we use cryo-electron microscopy (EM) and solid-state NMR spectroscopy to analyze the structure of the fibrils. These fibril structures are relevant for disease because they give rise to tissue damage and underlie the spreading of disease and protein misfolding inside the tissue. The two major techniques employed in this project, cryo-EM and solid-state NMR, are perfectly complementary. Cryo-EM can determine the overall electron density, symmetry and protofilament substructure and is amenable to body-derived fibrils. Solid-state NMR is an ideal tool to characterize dynamic regions within the aggregate. In this project we want to complementarily apply these techniques to study the structure of pathogenically relevant AA amyloid fibrils by examining bona fide fibrils from murine spleen, cell culture derived and in vitro formed fibrils.

在所有蛋白质错误折叠疾病中，AA淀粉样变性呈现出研究致病性淀粉样蛋白的结构，形成和传播的独特优势。在体外，细胞培养和小鼠模型系统中有高度相关的，可以重现淀粉样蛋白原纤维形成的关键步骤。淀粉样蛋白沉积物相对较大，可用于生化分析。鼠SAA1是一种细胞外蛋白质，足够小，可用于NMR。这些特征使我们能够采用一种综合的结构生物学方法，其中我们使用冷冻电子显微镜（EM）和固态NMR光谱法来分析原纤维的结构。这些原纤维结构与疾病有关，因为它们会导致组织损伤，并构成疾病和蛋白质内折叠式折叠的基础。该项目中使用的两种主要技术Cryo-EM和固态NMR是完全互补的。冷冻EM可以确定总的电子密度，对称性和原丝亚条件，并且可以适合身体衍生的原纤维。固态NMR是表征骨料内动态区域的理想工具。在这个项目中，我们希望通过检查鼠脾脏，细胞培养和体外形成的原纤维的良好原纤维来互补地应用这些技术来研究病原相关的AA淀粉样蛋白原纤维的结构。