Aging of the subcellular muscle proteome

描述（由申请人提供）：年龄相关的氧化损伤对蛋白质组的特异性在很大程度上是未知的。我们的长期目标是开发新的蛋白质组学策略，能够识别易受这种损伤的蛋白质。在这个探索性的R21应用中，我们的短期目标是使用亚细胞蛋白质组，羰基免疫捕获和ICAT技术的概念来识别（和发现）肌肉线粒体中与年龄相关的羰基相关蛋白。我们将使用Fisher 344大鼠作为动物模型。我们将比较两种肌肉的蛋白质组，它们的功能随着年龄的增长而下降，比目鱼肌和半膜肌。虽然比目鱼肌显著依赖于线粒体产生ATP，但半膜肌依赖于糖酵解作为ATP的来源。因此，假设不同的ROS产生模式将导致这两种肌肉中不同的蛋白质氧化损伤。建议进行两项研究：（1）直接比较6月龄和28月龄两种肌肉类型中的羰基相关蛋白;（2）对同一动物在三个不同年龄的肌肉取样进行纵向研究。第二项研究中的样本限制将通过使用FT-ICR-MS和CE-LIF解决。

DESCRIPTION (provided by applicant): The specificity of age-related oxidative damage to proteomes is largely unknown. Our long-term goal is to develop new proteomic strategies capable of identifying proteins susceptible to such damage. In this exploratory R21 application, our short-term goal is to use the concept of subcellular proteomes, carbonyl immuno-capture, and ICAT technology to identify (and discover) age-related carbonyl-associated proteins in muscle mitochondria. We will use the Fisher 344 rat as an animal model. We will compare the proteomes of two muscles whose functions decline differently with aging, the soleus and semimembranosus. While the soleus is significantly dependent on mitochondria for ATP production, the semimembranosus relies on glycolysis as a source of ATP. As a result, it is hypothesized that different ROS production patterns will cause different protein oxidative damage in these two muscles. Two studies are proposed: (1) direct comparison of carbonyl-associated proteins in the two muscle types at 6 and 28 months of age; (2) a longitudinal study with muscle sampling from the same animal at three different ages. Sample limitations in the second study will be addressed by using FT-ICR-MS and CE-LIF.