Molecular and Cellular Analyses of Parkin Function

帕金功能的分子和细胞分析

• 批准号: 6773154
• 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/6773154
• 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）：假设通过泛素蛋白酶体途径（UPP）降解有缺陷的蛋白质降解已被认为在神经退行性疾病（如帕金森氏病（PD））中起着核心作用。推定的泛素连接酶成分Parkin中的突变会导致以中脑多巴胺神经元丧失为特征的家族性常染色体隐性形式。因此，已经假设，在常染色体隐性帕金森氏病中多巴胺神经元丧失的基础是帕金泛素化底物的效率低下和随之而来的毒性积累。我们进一步假设帕金可能在调节中枢神经系统的神经元生存中发挥直接作用。我们建议使用分子和细胞工具来研究蛋白质泛素化和神经元存活中Parkin作用的机制。我们的初步数据表明，Parkin与先前表征的泛素连接酶成分，F-box/WD重复含有蛋白HSEL-10和Cullin-1（CUL 1）相关联的多蛋白泛素连接酶复合物。此外，HSEL-10可将这种复合物引导到特定的底物，包括Cyclin E（神经元细胞凋亡的推定调节剂）。我们将检验以下假设：（1）Parkin泛素蛋白连接酶复合物的辅助成分用于调节或针对这项活动，并且（2）在Parkin相关的家族性PD中，过早的神经元死亡是泛素化和神经凋亡复杂型park蛋白相关的parde蛋白相关的泛素不良的结果。