CHARACTERIZATION OF POLYGLUTAMINE AGGREGATES BY MASS SPECTROMETRY

通过质谱法表征聚谷氨酰胺聚集体

• 批准号: 7955936
• 负责人: Michael Y Sherman
• 金额: $ 4.72万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-01 至 2010-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7955936
• 关键词: Affect; Biology; Cell Death; Cells; Computer Retrieval of Information on Scientific Projects Database; Data; Disease; Doctor of Philosophy; Funding; Grant; Huntington protein; Inherited; Institution; Journals; Lead; Manuscripts; Mass Spectrum Analysis; Medicine; Methods; Modeling; Neurodegenerative Disorders; Neurons; Procedures; Protein Isoforms; Proteins; Proteomics; Publishing; Research; Research Personnel; Resources; Source; United States National Institutes of Health; Yeasts; mutant; novel; polyglutamine; protein aggregate; protein protein interaction; research study

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Polyglutamine diseases are inherited neurodegenerative diseases caused by the expansion of polyglutamine tract in the disease-causing mutant proteins. A unifying pathological feature of polyglutamine disorders is the presence of microscopically discernible intracellular inclusions in the affected neurons. It has been proposed that expanded polyglutamine tract formed-aggregates participate in inappropriate protein-protein interactions that lead to cell death. To date, there is still no reliable method to isolate highly purified aggregates which are usually morphologically heterogeneous. We have developed a new purification method and the proteomics procedure was carried out to analyze the aggregate-interacting proteins by mass spectrometry. Mass spectrometry provides a powerful and direct analysis of aggregate proteins in tiny quantities and will help us understand the nativity of aggregates in the polyglutamine disorders. We have obtained mass spectral data that indicates the presence of several expected proteins in the aggregate and others that would not have been easily predicted. Immunochemical approaches have confirmed these aggregate interacting proteins. A manuscript describing our novel purification method was recently published. Recent experiments have provided evidence that the yeast cells form aggresomes from some isoforms of the Huntington proteins; this is the first indication that aggresomes can form in yeast. The manuscript describing the identification of proteins that are required for aggresome formation and presenting our new model for aggresome formation has been published in FASEB journal. Extensive details are included in the PhD thesis of Y. Wang, completed in June 2008.

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 多聚谷氨酰胺病是由致病突变蛋白中多聚谷氨酰胺链扩增引起的遗传性神经退行性疾病。多聚谷氨酰胺疾病的一个统一的病理特征是在受影响的神经元中存在显微镜可识别的细胞内包涵体。 已经提出，扩展的多聚谷氨酰胺束形成的聚集体参与导致细胞死亡的不适当的蛋白质-蛋白质相互作用。迄今为止，仍然没有可靠的方法来分离高度纯化的聚集体，这些聚集体通常在形态上是异质的。我们开发了一种新的纯化方法，并进行了蛋白质组学程序，通过质谱分析聚集体相互作用蛋白。质谱法提供了一个强大的和直接的分析聚集蛋白在微量，并将帮助我们了解聚集体的多聚谷氨酰胺疾病的诞生。 我们获得的质谱数据表明聚集体中存在几种预期的蛋白质以及其他难以预测的蛋白质。免疫化学方法已经证实了这些聚集相互作用的蛋白质。最近发表了一份描述我们新的纯化方法的手稿。最近的实验提供了证据表明，酵母细胞从亨廷顿蛋白的某些同种型形成攻击体;这是第一个表明攻击体可以在酵母中形成的迹象。描述了侵略性基因组形成所需的蛋白质的鉴定并提出了我们的侵略性基因组形成新模型的手稿已发表在FASEB杂志上。在Y.王先生，于二零零八年六月完成。