Isolation and some properties of mitochondria from the abdomen muscle of the crayfish Orconectes limosus

Abstract

1. 1. Procedure for isolation of functionally and morphologically intact mitochondria from the abdomen muscle of the crayfish Orconectes limosus is described. 2. 2. Mitochondria obtained by this method show high rates of oxidation of succinate, malate, oxoglutarate, fumarate and pyruvate plus oxaloacetate; exhibit high respiratory control, and ADP:O ratios correspond to the theoretical value. 3. 3. Glutamate alone is oxidized slowly, but addition of oxaloacetate stimulates this oxidation. Other substrates as pyruvate alone, oxaloacetate alone, isocitrate, citrate, α-glycerophosphate and β-hydroxybutyrate are oxidized very poorly by these mitochondria, which are also able to oxidize reduced cytochrome c but not at all NADH. 4. 4. Oxidation of cytochrome c is only slightly stimulated by solubilizing agents Triton X-100 and deoxycholate. Triton X-100 at concentrations above 0.01% inhibited oxidation of reduced cytochrome c. 5. 5. The problem of outer mitochondrial membrane permeability for cytochrome c is discussed. 6. 6. The presence of arginine kinase activity was detected in the crayfish muscle mitochondria.