Isolation of totally inverted submitochondrial particles by sonication of beef heart mitochondria.
A novel procedure for isolating totally inverted preparations of submitochondrial particles by sonication of beef heart mitochondria is described. The procedure involves only differential centrifugation in 0.25 M sucrose containing 0.15 M KCl. The submitochondrial particles have 96% of their cytoplasmic face cytochrome c-binding sites sequestered within the particles. Mild sonication exposes cytochrome c-binding sites to the medium. The oligomycin-sensitive ATPase of sonic-derived submitochondrial particles, like that of electron transport particles, is inhibited 98% by exogenous isolated ATPase inhibitor protein. NADH oxidase activity in these particles is inhibited by oligomycin. The respiratory control index (uncoupled rate/oligomycin-inhibited rate) is approximately 3.4 and can be increased by washing the particles with medium containing bovine serum albumin.
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000818:Animals; D002417:Cattle; D002458:Cell Fractionation / Q000379:methods*; D003574:Cytochrome c Group / Q000378:metabolism; D004579:Electron Transport; D066298:In Vitro Techniques; D008929:Mitochondria, Heart / Q000378:metabolism / Q000648:ultrastructure*; D009097:Multienzyme Complexes / Q000378:metabolism; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism; D010084:Oxidation-Reduction; D011506:Proteins / Q000378:metabolism; D013010:Sonication; D014465:Ultrasonics
D003574:Cytochrome c Group; D009097:Multienzyme Complexes; D011506:Proteins; C023107:NADH oxidase; D009247:NADH, NADPH Oxidoreductases; D000251:Adenosine Triphosphatases