Figure S1 - Mitochondrial Aminopeptidase Deletion Increases Chronological Lifespan and Oxidative Stress Resistance while Decreasing Respiratory Metabolism in S. cerevisiae

Blue Native Gel Separation of mitochondrial respiratory supercomplexes from icp55Δ and BY4741 yeast strains cultured in glycerol containing media. The parental yeast strain BY4741 and the icp55Δ strain were cultured in glycerol containing media restricting energy metabolism to oxidative phosphorylation. Mitochondria were isolated with differential centrifugation, digitonin-solubilized and separated on a 4–16% bis-tris gel using blue native gel electrophoresis. Two replicates of each strain were examined and demonstrated comparable abundance of the respiratory complexes, labeled at the right-hand side of the second gel (arrowheads designate bands containing complex V components and asterisks designate complex III dimers with complex IV monomers or dimers) (A). The migration pattern of respiratory complexes from digitonin solubilized mitochondria of the parental strain (BY4741) were characterized by separation on a 4–16% bis-tris gel with molecular weight standards (Stand) and coomassie staining (B). In-gel activity assays of complex IV (CIV) labeled the complex III dimers with complex IV monomers or dimers with a pale brown precipitate, indicated by asterisks (B). In-gel activity assays of complex V on a light background (CV light) and dark background (CV dark) labeled complex V dimers and monomers with a white precipitate, indicated by arrowheads (B). (EPS)