TURNOVER RATES IN THE PROTEIN POOL OF C ELEGANS WITH EXTREME LIFESPAN

寿命极长的线虫蛋白质库的周转率

• 批准号: 8365487
• 负责人: RENE A BRAECKMAN
• 金额: $ 2.87万
• 依托单位: BATTELLE PACIFIC NORTHWEST LABORATORIES
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8365487
• 关键词: Aging; Animals; Biological Models; Biology; Caenorhabditis elegans; Collaborations; Food; Funding; Genetic; Grant; Insulin; Insulin-Like Growth Factor Receptor; Label; Life; Longevity; Maintenance; Mass Spectrum Analysis; Methods; National Center for Research Resources; Physiologic pulse; Principal Investigator; Proteins; Proteomics; Research; Research Infrastructure; Resolution; Resources; Sampling; Signal Transduction; Source; Technology; Translation Initiation; United States National Institutes of Health; cost; dietary restriction; experience; mutant; protein degradation

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. With its short lifespan, genetic amenability and the availability of long-lived mutant strains, C. elegans has become one of the most attractive model systems for aging research. In a previous collaboration, we have discovered very intriguing proteomic changes in worms that are long-lived due to dietary restriction or knockdown of insulin/IGF-like signaling. An important part of these changes is related to protein turnover. We hypothesize that lifespan extension in C. elegans is supported by changes in the turnover of specific protein pools. In the proposed project, we will apply the sensitive and reliable method pulsed SILAC, to verify whether proteins that are important for somatic maintenance are turned over more rapidly in long-lived worms compared to the corresponding controls. On the other hand, we expect a compensatory decrease in the turnover of the bulk of other proteins in these long-lived animals. For the experimental setup, we will use two mutant strains with extended lifespan: daf-2 (which is an insulin/IGF receptor mutant) and ife-2 (a somatic translation initiation mutant) and one strain in which lifespan is extended by environmental manipulation and dietary restriction by food dilution. These three long-lived strains will be compared to a corresponding control strain with normal lifespan. Worm culturing, SILAC labeling and sampling will be performed in the Braeckman lab, known for its extensive experience with C. elegans culturing and aging studies. These samples will be subsequently analyzed and quantified with state-of-the-art world class technology utilizing high resolution and mass accuracy mass spectrometry technologies at PNNL.

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 由于其寿命短，遗传上的顺从性和长寿突变株的可用性，C。线虫已成为衰老研究中最具吸引力的模型系统之一。在之前的合作中，我们发现了蠕虫中非常有趣的蛋白质组变化，这些变化是由于饮食限制或胰岛素/IGF样信号的敲低而导致的长寿。 这些变化的一个重要部分与蛋白质周转有关。 我们推测C. elegans是由特定蛋白质库的周转变化支持的。 在拟议的项目中，我们将采用灵敏可靠的脉冲SILAC方法，以验证与相应的对照相比，对体细胞维护重要的蛋白质在长寿蠕虫中是否更快地翻转。 另一方面，我们预计在这些长寿动物中，大部分其他蛋白质的周转量会出现补偿性下降。 对于实验设置，我们将使用两种具有延长寿命的突变菌株：daf-2（胰岛素/IGF受体突变体）和ife-2（体细胞翻译起始突变体），以及一种通过环境操作和食物稀释的饮食限制延长寿命的菌株。 将这三种长寿菌株与具有正常寿命的相应对照菌株进行比较。 蠕虫培养、SILAC标记和采样将在Braeckman实验室进行，Braeckman实验室以其对C. elegans培养和老化研究。 随后将在PNNL采用最先进的世界级技术，利用高分辨率和质量准确度质谱技术对这些样品进行分析和定量。