PIGEON MUSCLE MITOCHONDRIA: ION CONCENTRATIONS, OXIDATIVE PHOSPHORYLATION, SWELLING, AND EXTRAPARTICULATE SPACE

Abstract

Metabolically active mitochondria were isolated from pigeon breast muscle in cold 0.45 M sucrose with ATP, EDTA, and tris buffer at pH 7.4. Fresh mitochondria were analyzed for sodium, potassium, nitrogen, iron, and water. They were morphologically intact and showed respiratory control, respiration being limited by the phosphate acceptor and stimulated by dinitrophenol. The respiratory rate was the same in the Warburg respirometer as that in the polarograph chamber; no effect of agitation was observed. The rates of oxygen consumption over short periods of time were 30–40 μ atoms per hour per mg N for α-keto-glutarate and glutamate, 16 for malate, and 1.3 for β-OH butyrate; except in the last case, satisfactory P/O ratios were obtained. A soluble, heat-stable muscle factor, not precipitated at 100,000 × G, stimulated respiration with each substrate but did not affect phosphorylation. From optical density measurements of muscle mitochondria they appeared to behave as moderately good osmometers; only slight swelling occurred during oxidative phosphorylation, but this increased somewhat when respiration ceased. The osmotic dead space was estimated at 60% of the mitochondrial volume. With polyvinylpyrrolidone the extraparticulate space was found to be 27.8 ± 1.8%.