Survey of Age-Associated Carbonylation of Brain Proteins

与年龄相关的脑蛋白羰基化的调查

基本信息

批准号：
6863557
负责人：
LASZLO PROKAI
金额：
$ 26.67万
依托单位：
UNIVERSITY OF FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-09-30 至 2005-07-31
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): Oxidative stress may be a major causal factor underlying the aging process. Protein carbonylation is a potential marker for age-related deleterious changes. The present study applies a proteomic approach to facilitate the elucidation of the biochemical mechanisms that cause decrements in brain function, as reflected by behavioral performance of aged mice in cognitive and psychomotor tasks. The main methodological focus of the studies is the development of an isotope-coded affinity-tag (ICAT) strategy to enable mechanistic and quantitative studies on protein carbonylation. The proposed studies will determine appropriate parameters for carbonyl-directed identification and differential quantification of aging-associated oxidation-sensitive proteins based on this ICAT strategy and using mass spectrometry. Results of proteome-wide survey of oxidatively induced carbonylation in the aging mouse brain and the identification of age-associated oxidation-sensitive proteins will be related to the severity of the age-associated attenuations in specific cognitive and psychomotor functions. To this end, specific aims include the synthesis of reagent(s) that enable differential proteomics studies on protein carbonylation and the development of methods for the determination of oxidatively induced carbonylation of brain proteins. Oxidative stress on a synaptosomal fraction from mouse brain will then be used to study the occurrence, pathways and extent of protein carbonylation in vitro. Ultimately, a correlation of in vivo carbonylation of specific aging-associated oxidation-sensitive proteins with age and behavioral impairment will be sought. Experimental design of this study will entail scoring groups of aged mice on a specific cognitive or psychomotor task for the proteomics survey with young mice used as a control. Specific brain regions and subcellular fractions to be targeted for identification of carbonylated proteins will be those showing age-associated oxidative stress as measured by various methods. The degree of association will be determined between performance in each task and the degree of specific carbonylation associated with individual aging-associated oxidation-sensitive proteins.

描述（由申请人提供）：氧化应激可能是衰老过程的主要因素。蛋白质羰基是与年龄有关的有害变化的潜在标记。本研究采用了一种蛋白质组学方法来促进阐明导致脑功能降低的生化机制，这是由老年小鼠在认知和精神运动任务中的行为表现所反映的。研究的主要方法论重点是制定同位素编码的亲和力标签（ICAT）策略，以实现蛋白质羰基化的机械和定量研究。拟议的研究将根据此ICAT策略和使用质谱法确定适当的参数，以确定羰基定向鉴定和与衰老相关的氧化敏感蛋白的差异定量。蛋白质组对氧化诱导的小鼠大脑羰基化的调查以及与年龄相关的氧化敏感蛋白的鉴定的结果与特异性认知和心理瘤功能中与年龄相关的衰减的严重程度有关。为此，具体目的包括合成试剂的合成，该试剂能够对蛋白羰基化的差异蛋白质组学研究以及确定氧化诱导脑蛋白羰基化的方法的发展。然后，将使用小鼠大脑突触分数的氧化应激来研究体外蛋白话的发生，途径和程度。最终，将寻求特定衰老相关氧化敏感蛋白与年龄和行为障碍的体内羰基化的相关性。这项研究的实验设计将需要在特定的认知或心理运动任务上进行蛋白质组学调查的年龄小鼠的评分小组，并使用用作对照的年轻小鼠进行评分。用于鉴定羰基蛋白的特定大脑区域和亚细胞级分是通过各种方法测量的年龄相关氧化应激的特定脑区域和含有年龄相关的氧化应激的蛋白质。将在每个任务中的性能与与单个与衰老相关的氧化敏感蛋白有关的特定羰基化程度之间确定的关联程度。