PROTEIN TARGETS OF OXIDATIVE DAMAGE DURING AGING

衰老过程中氧化损伤的蛋白质目标

• 批准号: 6532521
• 负责人: RAJINDAR S SOHAL
• 金额: $ 28.74万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-08-01 至 2004-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6532521
• 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

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在细胞功能中导致衰老相关的损失的机制。假设：要检验的特定假设是“对特定蛋白质的氧化损害的应计造成细胞功能中与衰老相关的损失的原因”。 要审查的主要思想是，对特定蛋白质的氧化损害决定了在衰老过程中发生有害功能改变的性质和进展率。具体目的：将在果蝇果蝇的飞行肌肉中对线粒体和胞质蛋白进行研究，因为苍蝇的飞行能力在衰老过程中逐渐下降。 特异性蛋白质表现出与年龄相关的氧化损伤增加的蛋白质，该蛋白质通过羰基化表示，催化活性的损失将被鉴定出来。氧化损伤对蛋白质靶标与衰老过程之间的因果关系将在转基因和突变蝇中进行测试，这些果蝇分别过表达和不强调编码易受氧化损伤的蛋白质的基因。 另外，将在转基因或遗传选择的长寿命和对照苍蝇之间进行氧化损伤的比较。意义：这项研究的结果应提供有关：（1）将氧化应激与衰老过程中生理功能损失联系起来的机制，从而对衰老的氧化应激假说的有效性进行了进一步的批判性检验。 （2）确定衰老过程中蛋白质氧化损伤的靶标和随之而来的代谢失败。新颖性：蛋白质氧化损伤是一种选择性而不是随机现象的想法是新的，并且挑战了当前的概念。 实验方法将采用新方法来定量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氧化与衰老过程的氧化之间的因果关系。