Ubiquitination/Receptor Signaling--Regulation by Parkin

泛素化/受体信号转导——Parkin 的调节

• 批准号: 6454825
• 负责人: Benjamin L Wolozin
• 金额: $ 5万
• 依托单位: LOYOLA UNIVERSITY CHICAGO
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-06-01 至 2005-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6454825
• 关键词: Lewy body; Parkinson's disease; actin binding protein; actins; alpha synuclein; antisense nucleic acid; apoptosis; autosomal recessive trait; biological signal transduction; cell line; cell migration; collagen; enzyme activity; enzyme induction /repression; gene mutation; green fluorescent proteins; human tissue; integrins; laboratory rat; laminin; ligase; oxidative stress; postmortem; site directed mutagenesis; ubiquitin

DESCRIPTION (Provided by Applicant): Mutations in the gene coding for Parkin cause a rare familial form of Parkinsonism, autosomal recessive juvenile Parkinsonism, that results in death of dopaminergic neurons in the substantia nigra. To understand how parkin causes disease, we need to understand the regulation and function of parkin. Our studies have lead us to investigate ubiquitination, which is a process that regulates protein degradation. We hypothesize that parkin regulates ubiquitination of other proteins in response to cellular contact with matrix proteins (such as collagen and laminin), and thereby controls regulation of the cytoskeleton and signal transduction by matrix proteins and their integrin receptors. Loss of parkin function could cause neurodegeneration by inhibiting matrix signaling and impairing maintenance of processes by neurons. Our preliminary data support this hypothesis by demonstrating that parkin-dependent ubiquitination is activated by cellular binding to matrix proteins. We have also identified parkin binding proteins that are associated with integrins. Conversely, cell lines that have reduced parkin expression (due to anti-sense parkin cDNA) decrease ubiquitination, retract processes upon cellular exposure to matrix proteins, and have abnormal signal transduction. The goal of this proposal is to investigate the regulation of ubiquitination by parkin (Aim 1), determine the role of parkin in regulating signaling in response to exposure of cells to matrix proteins (Aim 2) and identify common functional deficits associated with disease-related mutations in parkin. Interestingly, parkin is also linked to other forms of neurodegeneration. Parkin binds to alpha-synuclein, and in brains from donors with Parkinson's disease parkin accumulates in inclusions that contain alpha-synuclein, and shows 75% less binding of parkin to two proteins, filamin and hCDCrel2a. We intend to investigate the mechanism of parkin dysfunction by determining how parkin function is altered in Lewy body diseases, and whether oxidation or alpha-synuclein aggregation causes the dysfunction of parkin (Aim 3). The research in this proposal will provide insight into the function of parkin, determine how mutations in parkin produce disease, and provide a new window to understand the molecular pathophysiology of Parkinson's disease.

描述（由申请人提供）：Parkin 编码基因突变 引起一种罕见的家族性帕金森病，常染色体隐性遗传青少年 帕金森症，导致体内多巴胺能神经元死亡 黑质。要了解帕金蛋白如何引起疾病，我们需要了解 Parkin的调节和功能。我们的研究引导我们调查 泛素化，这是调节蛋白质降解的过程。我们 假设 Parkin 调节其他蛋白质的泛素化反应 细胞与基质蛋白（例如胶原蛋白和层粘连蛋白）的接触，以及 从而控制细胞骨架和信号转导的调节 基质蛋白及其整合素受体。帕金功能丧失可能 通过抑制基质信号传导和损害引起神经退行性变 神经元维持过程。我们的初步数据支持这一点 通过证明 Parkin 依赖性泛素化被激活来提出假设 通过细胞与基质蛋白结合。我们还鉴定了parkin结合 与整合素相关的蛋白质。相反，具有 Parkin 表达减少（由于反义 Parkin cDNA） 泛素化，在细胞暴露于基质蛋白时收缩过程， 并有异常的信号转导。该提案的目标是 研究parkin对泛素化的调节（目标1），确定 Parkin 在调节细胞暴露的信号传导中的作用 基质蛋白（目标 2）并识别与相关的常见功能缺陷 Parkin 中与疾病相关的突变。有趣的是，parkin 还与 其他形式的神经变性。 Parkin 与 α-突触核蛋白结合，并且在 患有帕金森病的捐赠者的大脑帕金在包裹体中积累 含有 α-突触核蛋白，并且 Parkin 与两个蛋白的结合减少 75% 蛋白质、细丝蛋白和 hCDCrel2a。我们打算研究其机制 通过确定路易体中帕金功能如何改变来确定帕金功能障碍 疾病，以及氧化或 α-突触核蛋白聚集是否会导致 Parkin 功能障碍（目标 3）。本提案中的研究将提供 深入了解 Parkin 的功能，确定 Parkin 突变如何产生 疾病，并为了解分子病理生理学提供新窗口 帕金森病。