Structural Studies of Mammalian Mitochondrial Ribosomes

描述（由申请人提供）：哺乳动物线粒体翻译系统是一个非常独特的系统，是所有系统中了解最少的系统之一。除了它们在蛋白质生物合成中的主要作用之外，哺乳动物线粒体核糖体还参与细胞凋亡和各种线粒体疾病状态。线粒体核糖体类似于细菌核糖体;然而，大约一半的线粒体核糖体蛋白在细菌和细胞质核糖体蛋白中没有任何同源物。拟议研究的主要重点是确定新发现的蛋白质在线粒体蛋白质合成机制中的作用。一些与细菌同源的蛋白质负责mRNA、tRNA和因子与核糖体的结合。许多关键的细菌核糖体蛋白在哺乳动物线粒体核糖体中缺失。利用蛋白质组学方法，在定位和配体结合实验中确定新的核糖体蛋白类替代缺失的细菌同系物。此外，我们已经发现了三种不同的变体S18（MRP-S18）的线粒体核糖体的小亚基使用蛋白质组学方法的存在。这一发现清楚地表明线粒体核糖体中存在三种构象不同的小亚基亚群。在这个提议中，我们建议研究一类新的核糖体蛋白和MRP-S18的三种不同变体在哺乳动物线粒体蛋白质生物合成中的作用和功能。此外，对这些蛋白质的翻译后修饰的研究将使我们能够更好地了解疾病状态和细胞凋亡中线粒体翻译机制的调节。

DESCRIPTION (provided by applicant): The mammalian mitochondrial translation system is a very unique system and is one of the least understood systems of all. Besides their primary roles in protein biosynthesis, mammalian mitochondrial ribosomes are involved in apoptosis and various mitochondrial disease states. Mitochondrial ribosomes resemble the bacterial ribosomes; however, approximately half of the mitochondrial ribosomal proteins do not have any homologs among the bacterial and cytoplasmic ribosomal proteins. The major focus of the proposed research is to determine the roles of newly identified proteins in protein synthesis machinery of the mitochondria. Some of the proteins with the bacterial homologs are responsible for mRNA, tRNA, and factor binding to ribosomes. A number of the crucial bacterial ribosomal proteins are missing in the mammalian mitochondrial ribosome. The new class ribosomal proteins replacing the missing bacterial homologs will be determined in the localization and ligand binding experiments using proteomics approaches. In addition, we have discovered the presence of three different variants of S18s (MRP-S18s) in the small subunit of mitochondrial ribosomes using a proteomics approach. This finding clearly implies the presence of three conformationally different sub-populations of small subunits in mitochondrial ribosomes. In this proposal, we are proposing to study roles and functions of the new class ribosomal proteins and the three different variants of MRP-S18s in the protein biosynthesis in mammalian mitochondria. Furthermore, studies on post-translational modifications of these proteins will enable us to better understand the regulation of the mitochondrial translational machinery in disease states and apoptosis.