Tau protein proteolysis signaling in Alzheimer's disease

阿尔茨海默病中的 Tau 蛋白水解信号

基本信息

批准号：
10728202
负责人：
Carol J. Huseby
金额：
$ 43.18万
依托单位：
ARIZONA STATE UNIVERSITY-TEMPE CAMPUS
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-08-15 至 2025-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10728202
关键词：
Affect Alzheimer&apos s Disease Amino Acid Sequence Apoptosis Appearance Axon Binding Biochemical Biological Assay Brain Brain Diseases Brain region Cell Culture Techniques Cell Nucleus Cell Survival Cell model Cell physiology Characteristics Clinical Cryoelectron Microscopy Cytoplasm Cytoskeleton Data Dementia Dependence Deposition Development Digestion Disease Dissociation Dyes Environment Event Filament Fostering Funding Future Growth Human In Vitro Incubated Intracellular Transport Kinetics Lasers Length Ligands Mammalian Cell Maps Measures Membrane Microscopic Microtubules Molecular Conformation Morphology Nature Neurodegenerative Disorders Neurons Pathogenesis Pathologic Pathology Pathway interactions Peptide Fragments Peptide Hydrolases Peptides Post-Translational Protein Processing Process Property Protein Fragment Protein Isoforms Proteins Proteolysis Rapid screening Reaction Recombinants Research Scheme Signal Transduction Site Staging Structure Tauopathies Testing Thermodynamics Toxic effect Transmission Electron Microscopy beta pleated sheet brain tissue light scattering monomer neuron component neuron loss new therapeutic target novel prion-like protein degradation response self assembly small molecule tau Proteins tau aggregation

项目摘要

Project Summary Pathologic Tau protein self-assembly templating and stereotypical regional spreading along neuronal networks correlates with loss of neurons and is central to clinical progression and staging in Alzheimer’s disease (AD). In diseased brain, Tau molecules begin to form filamentous aggregates with a rigid β-sheet rich core. Although some Tau protein fragments can spontaneously aggregate, recombinant full-length Tau produced in vitro or when expressed in mammalian cells does not aggregate spontaneously even at supersaturation. To initiate Tau protein to form aggregates, some unknown energy barrier must be overcome. Research shows that inducing Tau monomer to form a nucleus leading to filamentous elongation can be a heterogenous association such as with a membrane, small molecules, or peptides stabilizing the misfolded species for subsequent elongation steps. Proteolysis is a process of protein degradation which also controls multiple cellular functions. Tau is a natively unfolded protein sensitive to protease digestion and the longest human brain isoform contains more than forty cleavage sites which leads to hundreds of possible Tau-derived peptides. Peptides have been shown to rapidly induce the misfolding and aggregation of tau in vitro and in mammalian cells. Here we propose to test and rank the propensity of Tau aggregation induction by each protease-cleaved Tau peptide after incubation with full-length recombinant Tau isoforms and subsequently capture aggregate gross morphology using transmission electron microscopy (TEM). Intracellular signaling activity by Tau peptides will be detected in cell culture to further characterize each peptide. Peptides inducing full-length Tau protein to aggregate or having active cellular signaling properties may implicate protease components dysregulated in disease. Because many known cleavage sites on Tau protein remain uncharacterized, this study aims to collect preliminary data for future funding and will begin groundwork for long-term research of proteolytic post- translational modification to identify those sites on Tau which may produce toxic peptides in neurons.

项目概要病理性 Tau 蛋白自组装模板和沿神经元网络的典型区域扩散与神经元损失相关，并且对于阿尔茨海默病 (AD) 的临床进展和分期至关重要。在患病的大脑中，Tau 分子开始形成具有富含 β-折叠的刚性核心的丝状聚集体。一些Tau蛋白片段可以自发聚集，体外产生的重组全长Tau或当在哺乳动物细胞中表达时，即使在过饱和状态下也不会自发聚集。 Tau蛋白要形成聚集体，必须克服一些未知的能量障碍。诱导 Tau 单体形成核，导致丝状伸长可能是一种异质关联例如用膜、小分子或肽来稳定错误折叠的物种以进行后续处理延伸步骤。蛋白水解是蛋白质降解的过程，也控制多种细胞功能。 Tau 是一种对蛋白酶消化敏感的天然未折叠蛋白质，最长的人脑亚型包含超过四十个切割位点可产生数百种可能的 Tau 衍生肽。研究表明，它可以在体外和哺乳动物细胞中快速诱导 tau 蛋白的错误折叠和聚集。提议对每种蛋白酶切割的 Tau 肽诱导 Tau 聚集的倾向进行测试和排序与全长重组 Tau 同工型孵育并随后捕获聚集体后使用透射电子显微镜 (TEM) 观察 Tau 肽的细胞内信号传导活性。在细胞培养物中进行检测，以进一步表征诱导全长 Tau 蛋白形成的肽。聚集或具有活跃的细胞信号传导特性可能意味着蛋白酶成分失调由于 Tau 蛋白上许多已知的裂解位点仍未被表征，因此本研究旨在收集未来资助的初步数据，并将开始为蛋白水解后的长期研究奠定基础翻译修饰以确定 Tau 上可能在神经元中产生有毒肽的位点。