Inhibiting fibrillation of Tau through changes in local intramolecular interactions

通过改变局部分子内相互作用抑制 Tau 纤维颤动

基本信息

批准号：
10463883
负责人：
Sofia Bali
金额：
$ 3.77万
依托单位：
UT SOUTHWESTERN MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-05-01 至 2024-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10463883
关键词：
Adopted Affinity Alzheimer&apos s Disease Amyloid Amyloid Fibrils Automobile Driving Binding Biological Biological Assay Biophysics Brain Cells Deposition Diagnostic Disease Disease Progression Engineering Ensure Environment Event Family Flow Cytometry Fluorescence Resonance Energy Transfer Frontotemporal Dementia Future Goals In Vitro Inherited Knowledge Lead Link MAPT gene Maps Mass Spectrum Analysis Measures Mediating Microtubule Stabilization Microtubules Modeling Modification Molecular Conformation Morphology Mutation Nature Neurodegenerative Disorders Onset of illness Peptides Persons Prevalence Property Protein Isoforms Proteins Resistance Sampling Shapes Structure Tauopathies Testing Therapeutic Toxic effect United States Variant amyloid formation amyloid structure atomic interactions base biomarker development crosslink design disease-causing mutation effective therapy experimental study flexibility molecular dynamics monomer novel therapeutics prevent protein function recruit tau Proteins tau aggregation tau conformation tau function tau mutation therapeutic evaluation thermostability

项目摘要

Project Summary/Abstract Neurodegenerative diseases such as Alzheimer's disease are characterized by the abnormal deposition of fibrillar aggregates in the brain. The microtubule-associated protein tau can form such assemblies and defines a diverse group of diseases termed tauopathies. The prevalence of tau fibrils strongly correlates with disease progression, with mutations that increase aggregation propensity directly linked to disease. Even so, tau is thermostable and does not form fibrils in vitro or in cells without specific inducers. The intrinsically disordered nature of tau protein allows the sampling of multivalent interactions, facilitating the biological activity of microtubule binding. However, there is a gap in knowledge of how tau structurally transitions from aggregation- resistant to an aggregation-prone conformation. This proposal aims to fill this gap in knowledge by studying local interactions involving amyloid motifs in the aggregation-prone repeat domain of tau. I will leverage the differences in fibril toxicity of two isoforms of tau to create a map of sequence properties that define the fibrillization properties of tau. I hypothesize that interactions between amyloid motifs and their surrounding sequence mediate tau aggregation propensity; therefore, stabilizing these interactions to form local structures can prevent fibril formation. In this proposal, I will identify atomic interactions in tau that modulate aggregation propensity using molecular dynamics simulations combined with peptide aggregation assay. Then I will engineer modified tau constructs that are predicted to stabilize the local structure. Lastly, I will determine the local structure of the stabilized tau species using cross-linking mass spectrometry. Then I will test whether the stabilized structure maintains flexibility in the microtubule-binding motifs through in vitro and in cell microtubule-binding assays. This integrative approach will create a map of stabilizing local interactions within a tau monomer to design aggregation-resistant conformations of tau that retain biological microtubule-binding activity. By understanding the biophysical basis behind early misfolding events in tau, this project can inform the future design of diagnostics and therapeutics that stabilize non-toxic tau species to treat the diverse family of tauopathies.

项目概要/摘要神经退行性疾病，例如阿尔茨海默氏病，其特征是异常沉积大脑中的纤维状聚集体。微管相关蛋白 tau 可以形成此类组件并定义称为tau蛋白病的多种疾病。 tau 原纤维的流行与疾病密切相关进展，增加与疾病直接相关的聚集倾向的突变。即便如此，tau 仍然是热稳定，在没有特定诱导剂的情况下，在体外或细胞中不会形成原纤维。本质上是无序的 tau 蛋白的性质允许对多价相互作用进行采样，从而促进其生物活性微管结合。然而，对于 tau 蛋白如何在结构上从聚集转变的知识存在差距。抵抗易于聚集的构象。该提案旨在通过研究当地的知识来填补这一知识空白涉及 tau 易聚集重复结构域中淀粉样蛋白基序的相互作用。我会利用差异研究两种 tau 亚型的原纤维毒性，创建定义原纤维化特性的序列特性图的 tau 蛋白。我假设淀粉样蛋白基序与其周围序列之间的相互作用介导 tau 聚集倾向；因此，稳定这些相互作用以形成局部结构可以防止纤维化形成。在这个提案中，我将确定 tau 中的原子相互作用，使用以下方法调节聚合倾向：分子动力学模拟与肽聚集测定相结合。然后我将设计修改后的 tau 预计可以稳定局部结构的构建体。最后，我将确定本地结构使用交联质谱法稳定 tau 物种。然后我会测试结构是否稳定通过体外和细胞内微管结合测定保持微管结合基序的灵活性。这综合方法将创建一个稳定 tau 单体内局部相互作用的图来设计保留生物微管结合活性的 tau 蛋白的抗聚集构象。通过了解 tau 早期错误折叠事件背后的生物物理学基础，该项目可以为未来的诊断设计提供信息以及稳定无毒 tau 蛋白物种以治疗不同家族的 tau 蛋白病的疗法。