Delineating protein-protein interaction network of hyperphosphorylated tau in tauopathies

描绘 tau 病中过度磷酸化 tau 的蛋白质-蛋白质相互作用网络

基本信息

批准号：
9181067
负责人：
Min-Hao Kuo
金额：
$ 24.14万
依托单位：
MICHIGAN STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-08-15 至 2018-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9181067
关键词：
Affinity Chromatography Alzheimer&apos s Disease Amyloid beta-Protein Axonal Transport Binding Binding Proteins Biochemical Biochemical Genetics Biological Markers Brain Catalysis Cell Extracts Cell membrane Clinical Trials Country Databases Development Diagnostic Drug Design Event Failure Filament Future Genetic study Goals Health Human Knowledge Lead Light Medical Methods Microtubules Mining Mission Modeling Molecular Molecular Target Nerve Degeneration Neurodegenerative Disorders Neurofibrillary Tangles Neuronal Dysfunction Neurons Pathology Patients Phosphorylation Post-Translational Protein Processing Proteins Proteomics Recombinants Shapes System Tauopathies Technology Therapeutic Time United States National Institutes of Health Work Yeasts base cDNA Library cytotoxicity data integration design drug development drug discovery effective therapy gain of function genetic approach hyperphosphorylated tau insight loss of function neuroblastoma cell neurofibrillary tangle formation neuron loss novel prion-like protein protein interaction research study tau Proteins tau function therapeutic development tool tool development translational study yeast two hybrid system

项目摘要

Alzheimer's disease (AD) currently attacks more than five million patients in the US. Without an effective treatment or preventative regime, AD patients in this country will be close to 14 million by 2050, with annual medical expenses topping 1.2 trillion US dollars. A major pathological hallmark for AD and other neurodegenerative disorders collectively called tauopathies is the neurofibrillary tangles (NFTs) composed of hyperphosphorylated tau protein (p-tau). Pathological hyperphosphorylation of tau interferes with the normal, axonal transport-related function of tau. Multiple lines of evidence also reveal a gain-of-function cytotoxicity of p-tau. The identification of the molecular targets of p-tau that underlie neurodegeneration will likely provide novel targets for the development of early diagnostic tools and therapeutics. In this project, we propose to use complementary approaches to perform the first systematic identification of proteins that interact preferentially with p-tau. In the biochemical approach, we will use recombinant p-tau produced by the PIMAX-Cat technology to isolate p-tau binding proteins from neuroblastoma cell extracts. In the genetic approach, we will use the tethered catalysis/yeast two-hybrid (TC/Y2H) system, which is designed to identify protein-protein interactions induced by a post-translational modification including phosphorylation, to screen for p-tau binding proteins from a human brain cDNA library. All candidate hits discovered from these two methods will be verified and characterized in neural cell-based co-purification experiments. Database mining and data integration will reveal the protein-protein interaction network centering upon p-tau, and lead to models for p-tau inflicted neurodegeneration.

目前，美国有超过 500 万患者患有阿尔茨海默病 (AD)。如果没有有效的如果采取治疗或预防措施，到 2050 年，该国 AD 患者将接近 1400 万，每年医疗费用突破1.2万亿美元。 AD 和其他疾病的主要病理标志神经退行性疾病统称为 tau 病，是由神经原纤维缠结 (NFT) 组成过度磷酸化 tau 蛋白 (p-tau)。 tau 病理性过度磷酸化会干扰 tau 的正常轴突运输相关功能。多项证据也揭示了功能获得性 p-tau 的细胞毒性。神经退行性变背后的 p-tau 分子靶标的鉴定将可能为早期诊断工具和治疗方法的开发提供新的靶点。在这个项目中，我们建议使用补充方法对蛋白质进行首次系统鉴定优先与 p-tau 相互作用。在生化方法中，我们将使用由 PIMAX-Cat 技术从神经母细胞瘤细胞提取物中分离 p-tau 结合蛋白。在遗传方面方法中，我们将使用系留催化/酵母双杂交（TC/Y2H）系统，该系统旨在识别由翻译后修饰（包括磷酸化）诱导的蛋白质-蛋白质相互作用，以从人脑 cDNA 文库中筛选 p-tau 结合蛋白。从这些中发现的所有候选命中两种方法将在基于神经细胞的共纯化实验中得到验证和表征。数据库挖掘和数据整合将揭示以p-tau为中心的蛋白质-蛋白质相互作用网络，并引领 p-tau 造成的神经变性模型。