Isolation of totally inverted submitochondrial particles by sonication of beef heart mitochondria |
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Authors: | H James Harmon |
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Institution: | (1) Departments of Physics and Zoology, Oklahoma State University, 74078 Stillwater, Oklahoma |
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Abstract: | 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 cytochromec-binding sites sequestered within the particles. Mild sonication exposes cytochromec-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. |
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Keywords: | Submitochondrial particles ATPase cytochromec |
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