A proteomic approach to identify substrates of the AAA+ mitochondrial proteases

鉴定 AAA 线粒体蛋白酶底物的蛋白质组学方法

基本信息

批准号：
9128054
负责人：
Leo J Pallanck
金额：
$ 23.18万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-09-01 至 2018-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9128054
关键词：
Accounting Aging Animal Model Antioxidants Autophagocytosis Biological Biological Models DNA Repair Enzymes Diabetes Mellitus Disease Drosophila genus Elderly Explosion Failure Family Fostering Foundations Functional disorder Future Genes Genetic Genetic study Goals Health Hereditary Spastic Paraplegia Human Individual Inherited Knowledge Malignant Neoplasms Measures Methods Mitochondria Mitochondrial Encephalomyopathies Mitochondrial Proteins Mutation Neurodegenerative Disorders PINK1 gene Parkinson Disease Pathogenesis Pathway interactions Peptide Hydrolases Perrault syndrome Play Prevention Process Proteins Proteomics Proton-Translocating ATPases Quality Control Reactive Oxygen Species Research Role Spinocerebellar Ataxias Stress Testing Ursidae Family Work Yeasts age related biological adaptation to stress cerebro-oculo-dento-auriculo-skeletal syndrome fly human disease in vivo mitochondrial dysfunction oxidation parkin gene/protein prevent protein degradation protein misfolding repaired stable isotope tool

项目摘要

DESCRIPTION (provided by applicant): Recent studies of the Parkinson's disease factors PINK1 and Parkin indicate that they play a critical role in the degradation of damaged mitochondria through a mitochondrial selective form of autophagy, termed mitophagy. This advance has fostered a greater appreciation of the importance of mitochondrial quality control to human health, and has led to a dramatic increase in research on mitophagy. However, mitophagy is only one of several mechanisms of mitochondrial quality control, and our recent work indicates that mitophagy accounts for less than half of all mitochondrial protein degradation in the model organism Drosophila. This unexpected finding suggests that non- mitophagic degradative processes are primarily responsible for mitochondrial protein quality control. We hypothesize from this and other findings that mitochondrial-resident proteases account for the majority of the protein degradative quality control that occurs in mitochondria. While genetic studies of the mitochondrial ATPase Associated with diverse cellular Activities (AAA+) family of proteases in yeast suggest that they play an important role in mitochondrial protein quality control, few of their substrates are known. Furthermore, the metazoan counterparts of these proteases have been little studied, despite the fact that mutations in the genes encoding several of them cause human disease. We propose to identify in vivo substrates of the four metazoan AAA+ mitochondrial proteases in Drosophila by using a stable isotope proteomic method to compare the half-lives of mitochondrial proteins in WT flies with those in flies that bear genetic perturbations targeting these proteases. Moreover, we will test whether these proteases degrade mitochondrial proteins that are damaged by two oft-cited mitochondrial stresses: oxidation and protein misfolding. Our work will advance the basic understanding of mitochondrial quality control by defining the biological roles of these proteases, and will provide a foundation to study the mechanisms by which mutations in the genes encoding these proteases cause disease.

描述（由适用提供）：对帕金森氏病因素PINK1和PARKIN的最新研究表明，它们通过线粒体选择性形式的自噬称为线粒体，在降解线粒体降解中起关键作用，称为线粒体。这一进步促进了对线粒体质量控制对人类健康的重要性的更大认识，并导致了线粒体研究的急剧增加。但是，线粒体只是线粒体质量控制的几种机制之一，我们最近的工作表明，线粒体占果蝇模型组织中所有线粒体蛋白降解的一半。这一意外发现表明，非线粒体降解过程主要负责线粒体蛋白质质量控制。我们从这一发现和其他发现中假设线粒体居住的蛋白质解释了线粒体中发生的大多数蛋白质降解质量控制。尽管酵母中与各种细胞活性（AAA+）蛋白家族相关的线粒体ATPase的遗传研究表明它们在线粒体蛋白质质量控制中起着重要作用，但很少有人知道它们的底物。此外，这些蛋白质的后生同生很少研究，这是一个事实，即编码其中几种的基因中的突变会引起人类疾病。我们建议通过使用稳定的同位素蛋白质组学方法比较果蝇中的四个后生AAA AAA+线粒体蛋白的体内底物，以比较WT蝇中线粒体蛋白的半衰弱与靶向这些蛋白质的蝇的蝇中的蝇蝇中的半角膜蛋白。此外，我们将测试这些蛋白是否会降解两种经常引用的线粒体胁迫受损的线粒体蛋白：氧化和蛋白质错误折叠。我们的工作将通过定义这些蛋白酶的生物学作用来提高对线粒体质量控制的基本理解，并将为研究编码这些蛋白酶引起疾病的基因突变的机制提供基础。