Solid-state NMR Structural Characterizations of Polymorphic Transthyretin Amyloids

多态性运甲状腺素蛋白淀粉样蛋白的固态 NMR 结构表征

• 批准号: 10164870
• 负责人: KWANG HUN LIM
• 金额: $ 21.91万
• 依托单位: EAST CAROLINA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10164870
• 关键词: Adopted; Alzheimer' s Disease; Amyloid; Amyloid beta-Protein; Amyloidosis; Biological; Biological Markers; Collaborations; Degenerative Disorder; Deposition; Disease; Docking; Exhibits; Goals; Human; Hydrophobicity; Label; Letters; Link; Molecular; Molecular Conformation; Mutation; Parkinson Disease; Pathogenicity; Pathway interactions; Phenotype; PrP; Prealbumin; Prion Diseases; Prions; Process; Proteins; Research; Resolution; Scheme; Structural Models; Structure; Testing; Therapeutic Agents; Tissues; Variant; alpha synuclein; amyloid formation; amyloid structure; beta pleated sheet; biophysical analysis; conformational conversion; disease phenotype; flexibility; human disease; improved; innovation; insight; interest; interfacial; intermolecular interaction; monomer; mutant; non-prion; polypeptide; prevent; programs; protein aggregation; protein folding; protein function; protein misfolding; simulation; solid state nuclear magnetic resonance; therapeutically effective; three dimensional structure; tissue degeneration; tool

Protein misfolding and amyloid formation is implicated in numerous diseases such as amyloidoses, prion and Alzheimer's diseases. Prion disease is unique in that the natively folded prion protein forms aggregates with distinct molecular conformations (prion strains), which underlie different disease phenotypes.1-3 The prion strain may be encoded in the primary sequence and mutations of the protein induce different strains, causing distinct disease phenotypes. Recent studies have suggested the strain hypothesis is applicable to other amyloid diseases that also manifest diverse disease phenotypes.1,2,4,5 Nonprion amyloids were shown to exhibit a wide conformational diversity,6-10 which may be linked to the phenotype variations. However, little is known about molecular basis of the diverse misfolding pathways and structural diversity of amyloid. Structural studies of the amyloid are essential to understanding molecular mechanism of amyloid diversity. Effect of the pathogenic mutations on misfolding pathway should also be examined. The systematic biophysical studies have, however, been challenging for previously investigated amyloidogenic proteins due to the limited number of pathogenic mutations associated with distinct disease phenotypes. In addition, the most extensively studied polypeptides, β-amyloid and α-synuclein associated with Alzheimer's and Parkinson's diseases respectively, are natively unfolded, rendering the polypeptides not amenable for mechanistic studies of the initial conformational transition (misfolding). This research program is aimed at investigating amyloid formation mechanisms of a natively folded protein, transthyretin (TTR), using solid-state NMR. Transthyretin (TTR) is one of more than 30 human proteins that undergo an aberrant conformational change and misassemble into β-structured amyloid. Amyloid formation of wild type and more than 100 mutant forms of TTR are known to cause various amyloidoses with enormous phenotype diversity.11 The main hypothesis of this proposal is that pathogenic mutant forms of TTR may have distinct misfolding pathways, adopting diverse amyloid conformations with different toxic activities, which may result in diverse disease phenotypes and tissue-selective depositions. The hypothesis will be tested through the structural characterization of amyloid derived from wild-type and various pathogenic mutant forms of TTR. In particular, solid-state NMR with innovative labeling schemes will provide valuable insights into amyloid diversity. Specific aims of the proposal are to explore: (1) Native-like structural features of amyloid core regions. (2) Conformational changes of the loop regions during amyloid formation. (3) Quaternary structure of WT and mutant forms of TTR amyloid. Mechanistic understanding of the misfolding and amyloid formation pathways would be critical to developing effective therapeutic strategies for TTR amyloidoses.

蛋白质不折叠和淀粉样蛋白形成在许多疾病中隐含，例如淀粉样蛋白， prion和阿尔茨海默氏症的疾病。 prion病是独一无二的 具有不同分子构象（prion菌株）的聚集体，这是不同的疾病 表型。1-3prion菌株可以在蛋白质的一级序列和突变中编码 诱导不同的菌株，导致不同的疾病表型。最近的研究表明 菌株假设适用于也表现出多种疾病的其他淀粉样蛋白疾病 表型。证明1,2,4,5非质淀粉样蛋白表现出广泛的构象多样性，6-10， 可能与表型变化有关。但是，关于分子基础知之甚少 淀粉样蛋白的多种错误折叠途径和结构多样性。淀粉样蛋白的结构研究是 了解淀粉样蛋白多样性的分子机制必不可少。病原 还应检查有关错误折叠途径的突变。系统的生物物理研究已经 然而，由于有限的 与不同疾病表型相关的致病突变数量。另外，最多 与阿尔茨海默氏症相关的广泛研究多肽，β-淀粉样蛋白和α-突触核蛋白 帕金森的疾病分别是本地展开的，使多肽不适合 初始会议过渡（错误折叠）的机械研究。该研究计划针对 在研究本内折叠蛋白的淀粉样蛋白形成机理时，使用甲状腺素蛋白（TTR），使用 固态NMR。硫代蛋白（TTR）是30多种人类蛋白质之一，这种蛋白质发生异常 构象变化和无质体成β结构淀粉样蛋白。野生型淀粉样蛋白形成和 已知有100多种突变体形式的TTR会引起各种淀粉样蛋白 表型多样性。11该提案的主要假设是TTR的致病突变体形式 可能具有明显的错误折叠途径，采用潜水员淀粉样蛋白构象 活动，可能导致潜水疾病表型和组织选择性沉积。 假设将通过源自野生型和淀粉样蛋白的结构表征来检验 TTR的各种致病突变形式。特别是具有创新标签的固态NMR 方案将为淀粉样蛋白多样性提供宝贵的见解。该提案的具体目的是 探索：（1）淀粉样蛋白核心区域的天然样结构特征。 （2）构象变化 淀粉样蛋白形成过程中的环区域。 （3）WT和突变形式的第四纪结构 淀粉样蛋白。对错误折叠和淀粉样形成途径的机械理解将是 对于制定有效的TTR淀粉样蛋白治疗策略至关重要。

项目成果

Toxic Misfolded Transthyretin Oligomers with Different Molecular Conformations Formed through Distinct Oligomerization Pathways.

• DOI: 10.1021/acs.biochem.2c00390
• 发表时间: 2022-11-01
• 期刊: BIOCHEMISTRY
• 影响因子: 2.9
• 作者: Dasari, Anvesh K. R.;Yi, Sujung;Coats, Matthew F.;Wi, Sungsool;Lim, Kwang Hun
• 通讯作者: Lim, Kwang Hun

CD and Solid-State NMR Studies of Low-Order Oligomers of Transthyretin.

• DOI: 10.1007/978-1-0716-2597-2_21
• 发表时间: 2023
• 期刊: Methods in molecular biology (Clifton, N.J.)