Mitochondrial Functional Proteomics

Mitochondria are intracellular organelles that regulate a number of vital processes in eukaryotic cells. They are not only the primary sites of energy production but are also the primary source of the harmful reactive Oxygen Species that cause oxidativedamage to the cell. In addition they regulate the critical process of programmed cell death (apoptosis). Thus proper mitochondrial function is critical to optimal metabolic performance and health. In order to understand the regulation mitochondrialfunction, and its role in human metabolism and disease it is imperative that we identify all the proteins that function in the mitochondria. In this proposal we detail a unique technology called CD-tagging (Central Dogma Tagging) which we will use tocomprehensively identify mitochondrial proteins. At the end of Phase I, we anticipate having a database of 50-100 mitochondrial genes and proteins. In addition, we will have a set of cell lines of equal size, each expressing a single mitochondrialprotein in tagged form. By the end of Phase II, the database should contain thousands of entries that collectively cover the majority of the mitochondrial proteins and the genes that encode them, and the cell line collection should be equally as large.Since the tags will reside in mouse ES cells, the potential will exist to generate transgenic animals expressing the tagged genes. These transgenic mouse lines will be invaluable for determining the functions of individual mitochondrial proteins and forexploring means to modulate mitochondrial stability and structure.