SUMMARY: Mitochondria were prepared in large quantities from by grinding hyphae with glass beads in an Eppenbach Micro-mill. Observation in the electron microscope indicated that some of the isolated mitochondria were probably damaged during preparation. Nevertheless, the crude mitochondrial fraction was capable of coupling phosphorylation to the oxidation of seven different substrates tested. The P: O ratio obtained with succinate ranged from 0.7 to 1.3 in different experiments with different mitochondrial preparations, but only low respiratory control ratios were observed. The effects of common inhibitors of oxidative phosphorylation were similar to those reported with mammalian mitochondria. In addition, ATPase and ATP-Pi exchange activities, as well as ion accumulation, were measured in the Neurospora mitochondria. The use of a ‘preparation’ medium containing sucrose, 0.25 ; EDTA, 0.005 (pH 7.0); bovine serum albumin (0.15%, w/v) was necessary to obtain these results.


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