PROTEIN TARGETS OF OXIDATIVE DAMAGE DURING AGING

衰老过程中氧化损伤的蛋白质目标

基本信息

批准号：
6637792
负责人：
RAJINDAR S SOHAL
金额：
$ 29.6万
依托单位：
UNIVERSITY OF SOUTHERN CALIFORNIA
依托单位国家：
美国
项目类别：
财政年份：
1999
资助国家：
美国
起止时间：
1999-08-01 至 2006-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6637792
关键词：
Drosophilidae SDS polyacrylamide gel electrophoresis aging cell senescence developmental genetics gene mutation genetically modified animals mitochondrial DNA oxidative stress protein purification protein structure function

项目摘要

The long-term goal of the proposed research is to understand the mechanisms by which oxidative stress causes senescence-associated losses in cellular functions. Hypothesis: The specific hypothesis to be tested is that "accrual of oxidative damage to specific proteins is responsible for the senescence-associated losses in cellular functions." The main idea to be scrutinized is that oxidative damage to specific proteins determines both the nature and the rate of progression of deleterious functional alterations occurring during the aging process. Specific aims: Studies will be conducted on mitochondrial and cytosolic proteins in the flight muscles of Drosophila melanogaster, as the flying ability of the flies gradually declines during aging. Specific proteins exhibiting an age-related increase in oxidative damage, indicated by carbonylation, and loss in catalytic activity will be identified. Causal association between oxidative damage to protein targets and the aging process will be tested in transgenic and mutant flies, which, respectively, overexpress and underexpress the genes encoding the proteins susceptible to oxidative damage. In addition, comparison of oxidative damage will be made between transgenic or genetically selected long-lived and control flies. Significance: Results of this study should provide important new knowledge about: (1) Mechanisms linking oxidative stress to the losses in physiological functions during aging and thus provide a further critical test of the validity of oxidative stress hypothesis of aging. (2) Identify targets of protein oxidative damage during aging and the consequent metabolic failures. Novelty: The idea that protein oxidative damage is a selective and not a random phenomenon is new and challenges the current concepts. The experimental approach will employ new methodology for the quantification of carbonylation of specific proteins as well as unique genetic strategies to test cause-and-effect relationships between the oxidation of specific proteins and the rate of the aging process.

拟议研究的长期目标是了解氧化应激导致细胞功能衰老相关损失的机制。假设：要测试的具体假设是“特定蛋白质氧化损伤的累积是与衰老相关的细胞功能丧失的原因”。需要仔细研究的主要思想是，特定蛋白质的氧化损伤决定了衰老过程中发生的有害功能改变的性质和进展速度。具体目标：研究果蝇飞行肌肉中的线粒体和胞质蛋白，因为果蝇的飞行能力随着衰老而逐渐下降。将鉴定表现出与年龄相关的氧化损伤增加（由羰基化表明）和催化活性丧失的特定蛋白质。蛋白质靶标的氧化损伤与衰老过程之间的因果关系将在转基因和突变果蝇中进行测试，这些果蝇分别过表达和低表达编码易受氧化损伤的蛋白质的基因。此外，还将对转基因或基因选择的长寿果蝇和对照果蝇之间的氧化损伤进行比较。意义：这项研究的结果应该提供以下重要的新知识：（1）氧化应激与衰老过程中生理功能丧失的联系机制，从而为衰老氧化应激假说的有效性提供进一步的关键检验。 (2) 确定衰老过程中蛋白质氧化损伤的目标以及随之而来的代谢衰竭。新颖性：蛋白质氧化损伤是一种选择性而非随机现象的观点是新的，对当前的概念提出了挑战。该实验方法将采用新的方法来量化特定蛋白质的羰基化，以及独特的遗传策略来测试特定蛋白质的氧化与衰老过程速率之间的因果关系。