Molecular and Cellular Analyses of Parkin Function

帕金功能的分子和细胞分析

• 批准号: 6917789
• 负责人: Asa Abeliovich
• 金额: $ 38.83万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-07-01 至 2008-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6917789
• 关键词: HeLa cells; Parkinson' s disease; SDS polyacrylamide gel electrophoresis; apoptosis; autosomal recessive trait; biological signal transduction; cell growth regulation; cellular pathology; clinical research; enzyme activity; gene mutation; immunocytochemistry; immunoprecipitation; laboratory mouse; ligase; molecular pathology; neural degeneration; neuroprotectants; protein degradation; protein structure function; ubiquitin; western blottings; yeast two hybrid system

DESCRIPTION (provided by applicant): Defective protein degradation through the ubiquitin proteasome pathway (UPP) has been hypothesized to play a central role in neurodegenerative disorders such as Parkinson's disease (PD). Mutations in Parkin, a putative ubiquitin ligase component, cause a familial, autosomal recessive form of PD characterized by midbrain dopamine neuron loss. It has therefore been hypothesized that inefficient degradation and consequent toxic accumulation of Parkin ubiquitination substrates underlie the loss of dopamine neurons in autosomal recessive Parkinson's disease. We further hypothesize that Parkin may play a direct role in regulating neuronal survival in the CNS. We propose to use molecular and cellular tools to investigate the mechanism of Parkin action in protein ubiquitination and neuronal survival. Our preliminary data indicate that Parkin associates in a multiprotein ubiquitin ligase complex with 2 previously characterized ubiquitin ligase components, the F-box/WD repeat-containing protein hSel- 10, and Cullin-1 (Cul 1). Furthermore, hSel-10 serves to direct this complex to specific substrates including Cyclin E, a putative regulator of neuronal apoptosis. We will test the hypotheses that (1) auxiliary components of the Parkin ubiquitin ligase complex serve to regulate or target this activity, and (2) that, in Parkin-associated familial PD, premature neuronal death is a consequence of defective ubiquitination and the accumulation of neuronal apoptosis-related Parkin complex substrates.

描述（由申请人提供）：据推测，通过泛素蛋白酶体途径（UPP）的有缺陷的蛋白质降解在帕金森氏病（PD）等神经退行性疾病中发挥着核心作用。 Parkin（一种假定的泛素连接酶成分）的突变会导致家族性、常染色体隐性遗传形式的 PD，其特征是中脑多巴胺神经元丢失。因此，有人推测，帕金泛素化底物的低效降解和随之而来的毒性积累是常染色体隐性遗传帕金森病中多巴胺神经元丧失的基础。我们进一步假设 Parkin 可能在调节中枢神经系统神经元存活中发挥直接作用。我们建议使用分子和细胞工具来研究 Parkin 在蛋白质泛素化和神经元存活中的作用机制。我们的初步数据表明，Parkin 在多蛋白泛素连接酶复合物中与 2 个先前表征的泛素连接酶成分、含有 F-box/WD 重复序列的蛋白 hSel-10 和 Cullin-1 (Cul 1) 结合。此外，hSel-10 还可将该复合物引导至特定底物，包括细胞周期蛋白 E（神经细胞凋亡的假定调节因子）。我们将检验以下假设：(1) Parkin 泛素连接酶复合物的辅助成分可调节或靶向该活性，(2) 在 Parkin 相关的家族性 PD 中，神经元过早死亡是泛素化缺陷和积累的结果。神经元凋亡相关的 Parkin 复合物底物。