CHARACTERIZATION OF POLYGLUTAMINE AGGREGATES BY MASS SPECTROMETRY

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 聚谷氨酰胺病是由致病突变蛋白中的聚谷氨酰胺束扩张引起的遗传性神经退行性疾病。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下可辨别的细胞内包涵体。有人提出，扩张的聚谷氨酰胺束形成聚集体，参与不适当的蛋白质-蛋白质相互作用，导致细胞死亡。到目前为止，仍然没有可靠的方法来分离高纯度的聚集体，这些聚集体通常在形态上是不同的。我们开发了一种新的纯化方法，并采用蛋白质组学方法对聚集相互作用的蛋白质进行了质谱分析。质谱学提供了一种强大而直接的微量聚集体蛋白分析方法，有助于我们了解聚谷氨酰胺疾病中聚集体的起源。我们已经获得了质谱学数据，表明聚集体中存在几个预期的蛋白质，以及其他不容易预测的蛋白质。免疫化学方法已经证实了这些聚集的相互作用蛋白。描述我们新的提纯方法的手稿最近出版了。最近的实验提供了证据，表明酵母细胞可以从亨廷顿蛋白的某些异构体形成侵袭体；这是第一个表明侵袭体可以在酵母中形成的迹象。

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.