Mitochondrial Functional Proteomics

Mitochondria are the organelles that play a central role in energy production in eukaryotic cells. Besides their bioenergetic function, mitochondria regulate cell death, modulate ionic homeostasis, oxidize carbohydrates and fatty acids, and participate inmultiple other catabolic and anabolic pathways. It is estimated that there are 1000-2000 distinct mitochondrial proteins, many of wich have not yet been identified. The invesigators propose to undertake a comprehensive mapping of the mitochondrial proteomeusing mouse heart as a tissue source. Mitochondria will be isolated, purified and separated using a variety of protocols developed and optimized in-house. Detection of low-abundance proteins will be facilitated by first fractionating the mitochondria intotheir various sub-compartments. The protein components of these subfractions will then be subjected to two complimentary mass spectrometry techniques. Data analysis using high-throughput informatics approaches will allow identification of these proteinswith high sensitivity.Furthermore, immunodepletion of high-abundance mitochondrial proteins (such as electron transport chain subunits) using specific antibodies will also be employed to identify mitochondrial proteins present at lower levels in these fractions. Identificationof novel mitochondrial proteins and pathways which are involved in human diseases connected with mitochondrial dysfunction.Elucidation of key mitchondrial intermediates involved in signaling pathways which modulate mitochondrial biogenesis and energy utilization, with a view to developing drugs or food supplements for enhancement of mitchondrial function and physicalperformance.Identification of mitochondrial proteins involved in apoptosis which could be regulated externally to attenuate the aging process.