Aging of the subcellular muscle proteome

亚细胞肌肉蛋白质组的衰老

• 批准号: 6950861
• 负责人: EDGAR A ARRIAGA
• 金额: $ 14.85万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-30 至 2007-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6950861
• 关键词: aging; biopsy; carbonyl compound; clinical research; free radical oxygen; intracellular; laboratory rat; liquid chromatography mass spectrometry; longitudinal animal study; mitochondria; oxidative stress; posttranslational modifications; protein quantitation /detection; proteomics; striated muscles

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.

描述(由申请人提供)：与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R21的这一探索性应用中，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。我们将比较比目鱼肌和半膜肌这两块肌肉的蛋白质组，这两块肌肉的功能随着年龄的增长而不同地衰退。比目鱼肌在很大程度上依赖线粒体来产生ATP，而半膜肌则依赖糖酵解作为ATP的来源。因此，假设不同的ROS产生模式会导致这两个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)从三个不同日龄的同一动物身上采集肌肉样本进行纵向研究。第二项研究中的样品限制将通过使用FT-ICR-MS和CE-LIF来解决。