Structural Characterization of hIAPP aggregates using MAS solid-state NMR

使用 MAS 固态 NMR 表征 hIAPP 聚集体的结构

• 批准号: 400866545
• 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: 2018
• 资助国家: 德国
• 起止时间: 2017-12-31 至 2020-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/400866545?language=en
• 关键词: Structural; Characterization; hIAPP; aggregates; using

The goal of the project is the determination of the structure of hIAPPox and hIAPPred fibrils at high resolution using MAS solid-state NMR. We will focus in particular on the N-terminal region of the peptide hormone to investigate the role of the disulfide bridge in the aggregate structure, and to understand its function in seeding and redox balancing. Seeding will be done with de novo generated fibrils as well as with ex vivo aggregate material. In addition to the pure hIAPPred and hIAPPox fibrils, we will produce hIAPP fibril samples that are obtained using a redox buffer that yields a 1:1 mixture of hIAPPred and hIAPPox in solution prior to aggregation. We aim to find out if only hIAPPred peptides are incorporated into the fibril, and how hIAPPox perturbs the fibril structure. In addition to static structures, we will investigate the dynamics of proteins in these amyloid fibrils, and aim to correlate local structural parameters to the thermodynamic stability of fibril structure. We expect that these experiments will allow to obtain a better mechanistic understanding of seeding and cross-seeding of an amyloid fibril.

该项目的目标是使用 MAS 固态 NMR 以高分辨率确定 hIAPPox 和 hIAPPred 原纤维的结构。我们将特别关注肽激素的 N 末端区域，以研究二硫键在聚集结构中的作用，并了解其在播种和氧化还原平衡中的功能。将用从头产生的原纤维以及离体聚集材料进行接种。除了纯 hIAPPred 和 hIAPPox 原纤维之外，我们还将生产 hIAPP 原纤维样品，这些样品是使用氧化还原缓冲液获得的，该缓冲液在聚集之前在溶液中产生 hIAPPred 和 hIAPPox 的 1:1 混合物。我们的目标是查明是否只有 hIAPPred 肽掺入原纤维中，以及 hIAPPox 如何扰乱原纤维结构。除了静态结构之外，我们还将研究这些淀粉样原纤维中蛋白质的动力学，并旨在将局部结构参数与原纤维结构的热力学稳定性相关联。我们期望这些实验能够更好地理解淀粉样原纤维的播种和交叉播种。