Molecular mechanisms of signal transduction by the mitochondrial life-span regulators p66Shc and Sirtuins

已经确定了几种与细胞衰老过程和相关疾病有关的蛋白质，包括Sirtuin蛋白和p66Shc。它们是复杂的信号网络的一部分，同步代谢适应和细胞凋亡等过程。哺乳动物有几种Sirtuin家族的NAD+依赖的蛋白脱乙酰酶的异构体。三种线粒体同工酶似乎有助于调节能量代谢和应激反应。同样，线粒体Shc亚型p66Shc作为应激感受器和细胞凋亡调节因子。这些信号系统及其联系的大部分分子细节仍有待确定，并可能揭示出治疗衰老相关疾病的有吸引力的干预点。我们的目标是鉴定和研究线粒体信号蛋白p66Shc、SIRT3和SIRT5的蛋白质/蛋白质相互作用，这些蛋白质/蛋白质相互作用有助于它们的定位、调节和底物识别。相互作用将通过亲和力实验来检测，底物和调节剂将通过活性分析来表征。将解决与多肽、蛋白质和小分子的络合物的晶体结构。这些相互作用将通过定点突变进行分析，我们将尝试利用这些信息来开发特定的调节剂。

Several proteins have been identified which are involved in cellular aging processes and related diseases, including Sirtuin proteins and p66Shc. They are part of a complex signaling network synchronizing processes such as metabolic adaptation and apoptosis.Mammals have several isoforms of the NAD+-dependent protein deacetylases of the Sirtuin family. Three mitochondrial isoenzymes appear to contribute to regulation of energy metabolism and stress response. Similarly, the mitochondrial Shc isoform p66Shc acts as stress sensor and apoptosis regulator. Most molecular details of these signaling systems and their connections remain to be identified and might reveal attractive intervention points for therapy of aging-related diseases. We aim to identify and study in molecular detail the protein/protein interactions of the mitochondrial signaling proteins p66Shc, Sirt3, and Sirt5, which contribute to their localization, regulation, and substrate recognition. Interactions will be detected through affinity experiments, and substrates and regulators characterized in activity assays. Crystal structures of complexes with peptides, proteins, and small molecules will be solved. The interactions will be analyzed by site-directed mutagenesis, and we will attempt to exploit this information for the development of specific modulators.