STABLE ISOTOPE LABELING METHOD DEVELOPMENT TO MEASURE PROTEIN TURNOVER IN A MUL

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 我们正在进行实验，使用稳定的同位素标记和质谱学来测量线虫的蛋白质周转。蛋白质的周转率是通过比较不同时间间隔内15N掺入蛋白质中的量来衡量的。这些实验是在所有菌株的背景下用成年GLP-4菌株进行的，以抑制蠕虫后代的生产，从而使所有动物处于相同的生命阶段。这种方法可以用来观察蛋白质在发育和其他过程中在多种生物中的周转情况。我们特别感兴趣的是测量线虫胰岛素/IGF-1信号通路中蛋白质的周转，同时过度表达该通路的配体。胰岛素/IGF-1信号通路中的蛋白质已被证明与长寿、代谢和发育有关。通过观察配体过表达过程中这一途径中蛋白质的周转情况，我们希望加深对胰岛素信号转导机制的理解。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. We are conducting experiments using stable isotope labeling and mass spectrometry to measure protein turnover in C. elegans. The turnover of protein is measured by comparing the incorporation of 15N into proteins over different time intervals. These experiments are being conducted with the adult glp-4 strain in the background of all strains to suppress worm progeny production and therefore keep all animals at the same life stage. This methodology can be used to look at the turnover of proteins in multiple organisms during development and other processes. We specifically are interested in measuring turnover of proteins in the insulin/IGF-1 signaling pathway in C. elegans while overexpressing the ligands of the pathway. Proteins in the insulin/IGF-1 signaling pathway have been shown to be involved in longevity, metabolism and development. By looking at the turnover of proteins in this pathway during ligand overexpression, we hope to improve our understanding of the mechanisms relating to insulin signaling.