Molecular mechanisms of transthyretin amyloidosis

运甲状腺素蛋白淀粉样变性的分子机制

基本信息

批准号：
10115719
负责人：
PETER Edwin WRIGHT
金额：
$ 45.64万
依托单位：
SCRIPPS RESEARCH INSTITUTE, THE
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-04-01 至 2024-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10115719
关键词：
Address Affect Age Aging Amyloid Amyloid Fibrils Amyloidosis Biological Assay Cardiomyopathies Complement DNA Sequence Alteration Data Deposition Dissociation Event Extracellular Protein Familial disease Goals Growth Heart Human Hybrids Impairment Individual Kinetics Lead Link Maps Measures Mediating Methods Molecular Molecular Conformation Molecular Weight Multidimensional NMR Techniques Multinuclear NMR Mutation Neurodegenerative Disorders Onset of illness Pathogenicity Pathway interactions Peptides Peripheral Nerves Physiological Play Population Prealbumin Process Proteins Research Role Seeds Shapes Structure Time Tissues Variant Virulence Factors X-Ray Crystallography age related aggregation pathway amyloid formation amyloidogenesis beta pleated sheet biophysical tools cold temperature cytotoxic design early onset experimental study extracellular globular protein human disease inhibitor/antagonist insight monomer mutant novel polymerization proteostasis shear stress small molecule

Address Affect Age Aging Amyloid Amyloid Fibrils Amyloidosis Biological Assay Cardiomyopathies Complement DNA Sequence Alteration Data Deposition Dissociation Event Extracellular Protein Familial disease Goals Growth Heart Human Hybrids Impairment Individual Kinetics Lead Link Maps Measures Mediating Methods Molecular Molecular Conformation Molecular Weight Multidimensional NMR Techniques Multinuclear NMR Mutation Neurodegenerative Disorders Onset of illness Pathogenicity Pathway interactions Peptides Peripheral Nerves Physiological Play Population Prealbumin Process Proteins Research Role Seeds Shapes Structure Time Tissues Variant Virulence Factors X-Ray Crystallography age related aggregation pathway amyloid formation amyloidogenesis beta pleated sheet biophysical tools cold temperature cytotoxic design early onset experimental study extracellular globular protein human disease inhibitor/antagonist insight monomer mutant novel polymerization proteostasis shear stress small molecule

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项目摘要

Many debilitating human diseases are associated with the extracellular misfolding and aggregation of globular proteins to form fibrillar deposits that are rich in β-sheet. There is considerable evidence that the process is initiated by local unfolding of the native structure to form aggregation-prone amyloidogenic intermediates. Aggregation then proceeds via nucleation-growth or downhill polymerization mechanisms. Despite their key role in amyloidogenesis, influencing the kinetic partitioning between aggregation and refolding pathways, very little is known about the structure of amyloidogenic intermediates because of their strong propensity to aggregate. The goals of the present proposal are to apply state-of-the-art NMR methods to elucidate the fundamental molecular events involved in transthyretin amyloidosis. Transthyretin amyloidosis is associated with numerous neurodegenerative diseases and cardiomyopathies. Misfolding and aggregation of transthyretin leads to fibrous deposits in the peripheral nerves and heart. Deposition of wild type protein is age related, whereas the familial diseases associated with genetic mutations that destabilize the quaternary and/or tertiary structure are early onset. The proposed research will provide novel insights into the fundamental molecular mechanisms by which wild type transthyretin aggregates and by which familial mutations destabilize the native transthyretin tetramer and drive the aggregation cascade. Real-time 19F NMR will be used to map the kinetic aggregation landscape of wild type and pathogenic variant transthyretin, characterize and quantify the population of intermediates that accumulate on the aggregation pathway, and examine mechanisms of inhibition by small molecules and peptides. Multidimensional NMR experiments will be utilized to elucidate the structure and dynamics of an alternate conformational state that promotes tetramer dissociation, and of cytotoxic monomeric and oligomeric intermediates, formed on the aggregation pathway, that promote aggregation and fibril growth. This research will advance our understanding of the underlying molecular events that initiate tetramer dissociation and promote entry into and progression down the aggregation cascade that leads to amyloid formation by both wild type human transthyretin and pathogenic variants.

许多使人类疾病衰弱的人与细胞外的错误折叠和聚集有关球状蛋白的形成富含β-折叠的原纤维沉积物。有大量证据该过程是通过本地结构的局部展开来启动的，以形成容易的聚合淀粉样蛋白生成中间体。然后聚集通过成核增长或下坡进行聚合机制。尽管它们在淀粉样生成中的关键作用，但影响动力学在聚集和重折叠途径之间进行分区，对结构的结构知之甚少淀粉样蛋白生成的中间体由于其强烈的聚集承诺。目标的目标目前的建议是应用最新的NMR方法来阐明基本分子经甲状腺素蛋白淀粉样变性涉及的事件。经硫代蛋白淀粉样变性与许多神经退行性疾病和心肌病。转蛋白的错误折叠和聚集导致周围神经和心脏中的纤维沉积物。野生型蛋白质的沉积是年龄相关，而与基因突变相关的家族疾病，使稳定稳定第四纪和/或三级结构是早期发作的。拟议的研究将提供新颖洞察野生型经硫代蛋白聚集体的基本分子机制而家庭突变破坏了本地甲状腺激素四聚体的稳定并驱动聚合级联。实时19F NMR将用于绘制动力学聚集局面野生型和致病性变体经甲状腺素素，表征和量化积聚在聚集途径上的中间体，并检查抑制的机制通过小分子和肽。多维NMR实验将用于阐明促进四聚体解离的替代构象状态的结构和动力学，以及在聚集途径上形成的细胞毒性单体和寡聚中间体的促进聚集和原纤维生长。这项研究将促进我们对启动四聚体解离并促进进入和的基本分子事件降低了导致两种野生型人类淀粉样蛋白形成的聚集级联经硫代蛋白和致病变体。