Dietary restriction, aging, and the proteasome

饮食限制、衰老和蛋白酶体

• 批准号: 7666085
• 负责人: Jeffrey Neil Keller
• 金额: $ 26.65万
• 依托单位: LSU PENNINGTON BIOMEDICAL RESEARCH CTR
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-01 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7666085
• 关键词: Age; Aging; Biochemical; Biogenesis; Biological Preservation; Cell Culture Techniques; Complex; Data; Exhibits; Impairment; In Vitro; Individual; Laboratories; Life; Longevity; Mammals; Modification; Pathway interactions; Peptide Hydrolases; Proteasome Inhibition; Proteins; Proteolysis; Proteome; Proteomics; Testing; age related; cell type; dietary restriction; in vivo; multicatalytic endopeptidase complex; normal aging; oxidation

DESCRIPTION (provided by applicant): The proteasome is an intracellular protease that is responsible for a significant amount of proteolysis, including the degradation of oxidized proteins and a large number of established short-lived proteins lacking oxidative modifications. Increasing evidence suggests that inhibition of the proteasome-proteolytic pathway occurs as a part of normal aging, with inhibition of proteasome function likely contributing to numerous age-related alterations in the proteome, including age-related increases in oxidized protein. However, the lack of a rigorous biochemical and proteomic analysis has made such estimations largely hypothetical and theoretical. Dietary restriction (DR) is the only known manipulation to consistently and reliably increase average and maximal lifespan in mammals. Numerous studies have demonstrated that DR can ameliorate age-related increases in oxidized protein, and suppress a variety of age-related alterations to the proteome. Data from our laboratory demonstrates that DR ameliorates age-related impairments in proteasome function, with the preservation of proteasome function possibly contributing to the beneficial effects of DR on the proteome. The focus of this proposal is to utilize rigorous proteomic and biochemical analysis to test the hypothesis that DR ameliorates age-related impairments in proteasome function as the result of its direct effects on the proteasome complex, with the preservation of proteasome activity contributing to DR-induced effects on the proteome (oxidized and non-oxidized proteins) during normal aging. The specific aims for testing this hypothesis are as follows: (1) To determine the effects of DR on proteasome subunit expression, proteasome biogenesis, proteasome composition, and proteasome oxidation; (2) To determine the effects of DR on the individual peptidase activities and protein degrading capabilities of the proteasome; (3) To determine the cell-type specific effects of impaired proteasome function on the proteome, including protein oxidation, in primary CNS cell cultures in vitro; (4) To determine if proteins which exhibit altered expression or increased oxidation following proteasome inhibition in vitro, exhibit similar changes during aging in vivo; (5) To determine if DR ameliorates alterations in the proteome, including increases in oxidized protein, observed during aging in vivo and following proteasome inhibition in vitro.

描述（由申请人提供）：蛋白酶体是一种细胞内蛋白酶，负责大量蛋白水解，包括氧化蛋白和大量缺乏氧化修饰的短寿命蛋白的降解。越来越多的证据表明，蛋白酶体蛋白水解途径的抑制是正常衰老的一部分，蛋白酶体功能的抑制可能导致蛋白质组中许多与年龄相关的改变，包括氧化蛋白的年龄相关增加。然而，由于缺乏严格的生物化学和蛋白质组学分析，这种估计在很大程度上是假设和理论的。饮食限制（DR）是唯一已知的操纵一致和可靠地增加哺乳动物的平均和最长寿命。许多研究表明，DR可以改善与年龄相关的氧化蛋白的增加，并抑制多种与年龄相关的蛋白质组改变。来自我们实验室的数据表明，DR改善了蛋白酶体功能中与年龄相关的损伤，蛋白酶体功能的保留可能有助于DR对蛋白质组的有益作用。该提案的重点是利用严格的蛋白质组学和生化分析来检验DR改善蛋白酶体功能中与年龄相关的损伤的假设，这是由于其对蛋白酶体复合物的直接影响，蛋白酶体活性的保留有助于在正常衰老期间DR诱导的对蛋白质组（氧化和非氧化蛋白质）的影响。本研究的主要目的是：（1）研究DR对蛋白酶体亚单位表达、蛋白酶体生物合成、蛋白酶体组成和蛋白酶体氧化的影响，（2）研究DR对蛋白酶体的肽酶活性和蛋白降解能力的影响，（3）研究DR对蛋白酶体的生物合成、蛋白酶体组成和蛋白酶体氧化的影响。（3）在体外原代培养的CNS细胞中，确定蛋白酶体功能受损对蛋白质组的细胞类型特异性影响，包括蛋白质氧化;（4）确定在体外蛋白酶体抑制后表现出改变的表达或增加的氧化的蛋白质在体内老化期间是否表现出类似的变化;（5）为了确定DR是否改善蛋白质组的改变，包括氧化蛋白的增加，在体内老化期间和体外蛋白酶体抑制后观察到。