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

基本信息

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
项目类别：
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&apos 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

项目摘要

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.

蛋白质错误折叠和淀粉样蛋白形成与许多疾病有关，例如淀粉样变性、朊病毒和阿尔茨海默氏病的独特之处在于，朊病毒蛋白形成了天然折叠。具有不同分子构象（朊病毒株）的聚集体，它们是不同疾病的基础表型1-3 朊病毒株可能编码蛋白质的一级序列和突变诱导不同的菌株，导致不同的疾病表型。菌株假说适用于也表现出多种疾病的其他淀粉样蛋白疾病表型。1,2,4,5 非朊病毒淀粉样蛋白表现出广泛的构象多样性，6-10 可能与表型变异有关，然而，人们对其分子基础知之甚少。淀粉样蛋白的不同错误折叠途径和结构多样性。对于理解淀粉样蛋白多样性的分子机制至关重要。还应该检查错误折叠途径的突变，系统的生物物理学研究，然而，由于有限的研究，先前研究的淀粉样蛋白形成一直具有挑战性。此外，与不同疾病表型相关的致病突变数量最多。广泛研究了与阿尔茨海默病相关的多肽、β-淀粉样蛋白和 α-突触核蛋白帕金森病分别是天然未折叠的，使得多肽不适合治疗该研究计划的目的是对初始构象转变（错误折叠）进行机制研究。研究天然折叠蛋白运甲状腺素蛋白 (TTR) 的淀粉样蛋白形成机制，使用固态 NMR 是 30 多种发生异常的人类蛋白质之一。构象改变并错误组装成野生型和β-结构淀粉样蛋白的形成。已知超过 100 种 TTR 突变形式可引起各种淀粉样变性表型多样性。11 该提案的主要假设是 TTR 的致病突变形式可能具有不同的错误折叠途径，采用具有不同毒性的不同淀粉样蛋白构象活动，这可能导致不同的疾病表型和组织选择性沉积。该假设将通过野生型和衍生型淀粉样蛋白的结构表征进行检验 TTR 的各种致病突变形式，特别是具有创新标记的固态 NMR。该计划将为淀粉样蛋白多样性提供有价值的见解。探索：（1）淀粉样蛋白核心区的类似天然结构特征（2）淀粉样蛋白核心区的构象变化。 (3) WT的四级结构和TTR的突变形式淀粉样蛋白的错误折叠和淀粉样蛋白形成途径的机制理解将是对于制定 TTR 淀粉样变性的有效治疗策略至关重要。

项目成果

期刊论文数量（5）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

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.