Survey of Age-Associated Carbonylation of Brain Proteins

与年龄相关的脑蛋白羰基化的调查

• 批准号: 7126850
• 负责人: LASZLO PROKAI
• 金额: $ 27.73万
• 依托单位: UNIVERSITY OF NORTH TEXAS HLTH SCI CTR
• 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 2009-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7126850
• 关键词: affinity chromatography; aging; behavior test; carbon; carbonyl compound; chemical synthesis; clinical research; cognition; electrospray ionization mass spectrometry; laboratory mouse; matrix assisted laser desorption ionization; neurophysiology; oxidative stress; phenylhydrazines; proteins; proteomics; psychomotor function; radiotracer; stable isotope; synaptosomes

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一个主要致病因素。蛋白质羰基化是与年龄相关的有害变化的潜在标志。本研究应用蛋白质组学方法来促进阐明导致大脑功能下降的生化机制，如老年小鼠在认知和精神运动任务中的行为表现所反映的那样。这些研究的主要方法论重点是开发同位素编码亲和标签（ICAT）策略，以实现蛋白质羰基化的机械和定量研究。拟议的研究将基于该 ICAT 策略并使用质谱法来确定适合老化相关氧化敏感蛋白的羰基定向鉴定和差异定量的参数。对衰老小鼠大脑中氧化诱导的羰基化的蛋白质组范围调查以及与年龄相关的氧化敏感蛋白的鉴定的结果将与特定认知和精神运动功能中与年龄相关的衰减的严重程度相关。为此，具体目标包括合成能够对蛋白质羰基化进行差异蛋白质组学研究的试剂，以及开发用于测定脑蛋白质氧化诱导的羰基化的方法。然后，小鼠大脑突触体部分的氧化应激将用于研究体外蛋白质羰基化的发生、途径和程度。最终，将寻求特定衰老相关氧化敏感蛋白的体内羰基化与年龄和行为障碍的相关性。这项研究的实验设计将需要对老年小鼠组的特定认知或精神运动任务进行评分，以进行蛋白质组学调查，并以年轻小鼠作为对照。用于鉴定羰基化蛋白质的特定脑区域和亚细胞部分将是那些通过各种方法测量显示出与年龄相关的氧化应激的区域和亚细胞部分。关联程度将取决于每项任务的表现和与各个衰老相关的氧化敏感蛋白相关的特定羰基化程度。