RP8: The neuronal interactome of mitochondrial m-AAA proteases

RP8：线粒体 m-AAA 蛋白酶的神经元相互作用组

• 批准号: 174794870
• 负责人: Professor Dr. Thomas Langer
• 金额: --
• 依托单位: Max-Planck-Institut für Biologie des Alterns
• 依托单位国家: 德国
• 项目类别: Research Units
• 财政年份: 2010
• 资助国家: 德国
• 起止时间: 2009-12-31 至 2014-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/174794870?language=en
• 关键词: RP8; neuronal; interactome; mitochondrial; AAA

Neurodegenerative disorders are often associated with a dysfunction of mitochondria, illustrating essential functions of mitochondria for neuronal survival. Mutations in m-AAA proteases, ATP-dependent proteolytic complexes in the inner membrane of mitochondria, cause distinct neurodegenerative disorders, but the molecular basis of these deficiencies are not understood. Studies in yeast identified crucial functions of m- AAA proteases for protein quality control and for the regulation of mitochondrial morphology and ribosome biogenesis. Here, we will define the neuronal interactome of m-AAA proteases, in order to identify critical substrates and cofactors that are required for neuronal maintenance. Transgenic mouse lines will be established allowing the inducible and tissue-specific expression of proteolytically inactive and dominant negative variants of m-AAA protease subunits using Cre-mediated recombination. Proteins copurifying with m-AAA protease complexes isolated from different regions of the brain will be identified and functionally characterized. Moreover, we will identify neuronal proteins processed by the m-AAA protease by determining the N-proteome of murine brain mitochondria isolated from wild type and m-AAA protease-deficient mice by mass spectrometry.

神经退行性疾病通常与线粒体功能障碍相关，说明线粒体对于神经元存活的基本功能。线粒体内膜中的m-AAA蛋白酶（ATP依赖性蛋白水解复合物）的突变引起不同的神经退行性疾病，但这些缺陷的分子基础尚不清楚。在酵母中的研究鉴定了m-AAA蛋白酶对于蛋白质质量控制和对于线粒体形态和核糖体生物发生的调节的关键功能。在这里，我们将定义m-AAA蛋白酶的神经元相互作用组，以确定神经元维护所需的关键底物和辅因子。将建立转基因小鼠系，允许使用Cre介导的重组诱导和组织特异性表达m-AAA蛋白酶亚基的蛋白水解失活和显性阴性变体。将鉴定与从脑的不同区域分离的m-AAA蛋白酶复合物共纯化的蛋白质并进行功能表征。此外，我们将通过质谱法确定从野生型和m-AAA蛋白酶缺陷小鼠中分离的小鼠脑线粒体的N-蛋白质组来鉴定由m-AAA蛋白酶加工的神经元蛋白。