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

本子项目是利用由NIH/NCRR资助的中心赠款提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是中心的，不一定是研究者的机构。我们正在进行实验，使用稳定同位素标记和质谱法来测量秀丽隐杆线虫的蛋白质周转。蛋白质的周转是通过比较15N在不同时间间隔内与蛋白质的结合来测量的。这些实验是在所有菌株的背景下以成年glp-4菌株进行的，以抑制蠕虫的后代产生，从而使所有动物保持在同一生命阶段。这种方法可以用来观察多种生物在发育和其他过程中的蛋白质周转。我们特别感兴趣的是测量秀丽隐杆线虫中胰岛素/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寿命、代谢和发育有关。通过观察配体过表达过程中该通路中蛋白质的转换，我们希望能提高我们对胰岛素信号传导机制的理解。

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.