Definition and characterization of the mitochondrial proteome of human cells (MitoCoP)

Aim of this project was the definition and characterization of the mitochondrial proteome of human cells, referred to as MitoCoP. To this end, we combined the complementary strengths of subtractive proteomics, ImportOmics and subcellular protein profiling in a global multidimensional classification approach. This multifaceted human mitochondria-centered analysis was expanded to a functional characterization of the mitochondrial proteome. The association of proteins with mitochondrial membranes was assessed by quantitative proteomic and single-protein studies. Protein copy numbers per mitochondrial unit were determined followed by a thorough compilation and functional classification of a high-quality human mitochondrial proteome. We further examined the organization of mitochondrial proteins in large functional assemblies by Blue Native (BN) complexome profiling and characterized functional protein networks of so far not described mitochondrial constituents by quantitative affinity purification mass spectrometry (q-AP-MS) and biochemical analysis. And finally, we systematically studied mitochondrial proteome dynamics and established a comprehensive subunit-resolved turnover map of large membrane protein machineries and central biosynthesis pathways of human mitochondria.