Modulating the fate of tau in the Hsp70 chaperone system.

调节 Hsp70 伴侣系统中 tau 蛋白的命运。

• 批准号: 8196724
• 负责人: Andrea Dooley Thompson
• 金额: $ 2.47万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-12-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8196724
• 关键词: ATP phosphohydrolase; Affect; Affinity; Aftercare; Alzheimer' s Disease; American; Amyloid beta-Protein; Binding; Binding Proteins; Biological Assay; Brain; Cell model; Cells; Chemicals; Client; Complex; Data; Enzyme-Linked Immunosorbent Assay; Genetic; Goals; Heat-Shock Proteins 70; Heat-Shock Response; Mass Spectrum Analysis; Mediating; Memory Loss; Microtubules; Modeling; Molecular; Molecular Chaperones; Molecular Conformation; Neurodegenerative Disorders; Nucleotides; Pattern; Positioning Attribute; Proteins; Proteolysis; Proteome; Recruitment Activity; Signal Transduction; Site-Directed Mutagenesis; Structure; Surface; Surface Plasmon Resonance; System; Tauopathies; Techniques; Testing; Work; base; inhibitor/antagonist; innovation; novel; peptide A; protein degradation; protein folding; protein misfolding; response; small molecule; tau Proteins; tau aggregation; therapeutic target; tool

DESCRIPTION (provided by applicant): The objective of this application is to understand how novel ATPase modulators of Hsp70 change the fate of its pathologically important client protein, tau. The central hypothesis states that ATPase modulation mediates the fate of tau by affecting which co-chaperones associate with Hsp70. Completion of this work will define the Hsp70 complexes that fate tau for refolding and degradation. Consequently, new and innovative ways to pharmacologically intervene in Alzheimer's disease (AD) and other tauopathies may be identified. The following aims will test the central hypothesis. Aim 1: Identify the co-chaperone composition of complexes induced by Hsp70 ATPase modulators in a cell- based model. The working hypothesis states that Hsp70 ATPase stimulators recruit a co- chaperone which promotes the accumulation of tau. Conversely, Inhibitors recruit a co-chaperone which promotes proteosomal degradation of tau. This hypothesis will be tested by performing an immunoprecipitatlon of tau, in both treated and untreated cells, followed by mass spectrometry. Aim 2: Determine the effect of ATPase modulators on IHsp70 binding interactions with co- chaperones and client protein tau. The working model postulates that Hsp70 ATPase modulation changes the binding affinity of certain co-chaperones for the Hsp70-tau complex. Surface plasmon resonance and an enzyme-linked immunosorbent assay will be used to test the effect of ATPase modulators on the formation of binary and ternary complexes between Hsp70, tau, and co-chaperones. Aim 3: Understand the structural basis for the effect of ATPase modulation on Hsp70, tau, co- chaperone complex formation. We predict that ATPase modulators cause Hsp70 to predominantly populate specific nucleotide bound conformations. Previous work, using partial proteolysis, has shown that distinctive cleavage patterns define the ATP and ADP conformations of Hsp70. Thus, this technique will be used to probe the structure of Hsp70 after treatment. Site-directed mutagenesis will also be used to define binding surfaces and allosteric networks affected by ATPase modulators. Relevance: More than five million Americans suffer from Alzheimer's disease (AD). In AD, misfolded proteins build up in the brain causing severe and irreversible memory loss. By therapeutically targeting a chaperone factor involved in the folding and degradation of misfolded proteins we may be able to take advantage of the body's own intrinsic system to treat AD.

描述（由申请人提供）：本申请的目的是了解HSP70的新型ATPase调节剂如何改变其病理上重要的客户蛋白Tau的命运。中央假设指出，ATPase调制通过影响哪个与HSP70相关的伴侣来介导Tau的命运。这项工作的完成将定义命运tau的HSP70复合物，以重新折叠和退化。因此，可以鉴定出药理学干预阿尔茨海默氏病（AD）和其他tauopathies的新的和创新的方法。以下目的将检验中心假设。 AIM 1：确定基于细胞模型中HSP70 ATPase调节剂诱导的复合物的共伴侣组成。工作假设指出，HSP70 ATPase刺激剂募集了促进TAU积累的共伴侣。相反，抑制剂募集了促进tau蛋白体降解的伴侣。该假设将通过在处理和未处理的细胞中进行tau的免疫原核来检验，然后进行质谱。 AIM 2：确定ATPase调节剂对IHSP70结合相互作用与共振子和客户蛋白tau的影响。工作模型假设HSP70 ATPase调制改变了某些共伴侣对HSP70-TAU复合物的结合亲和力。表面等离子体的共振和酶联免疫吸附测定法将用于测试ATPase调节剂对HSP70，TAU和共伴侣之间二元和三元复合物形成的影响。 AIM 3：了解ATPase调制对HSP70，TAU，共振子络合物复合物的影响的结构基础。我们预测ATPase调节剂会导致HSP70主要填充了特定的核苷酸结合构象。使用部分蛋白水解的先前工作表明，独特的切割模式定义了HSP70的ATP和ADP构象。因此，该技术将用于探测治疗后HSP70的结构。定点诱变也将用于定义受ATPase调节剂影响的结合表面和变构网络。相关性：超过500万美国人患有阿尔茨海默氏病（AD）。在AD中，错误折叠的蛋白质在大脑中积聚，导致严重和不可逆的记忆力丧失。通过治疗靶向涉及错误折叠蛋白折叠和降解的伴侣因子，我们可能能够利用人体自己的内在系统来治疗AD。