The measurement of the rotenone-sensitive NADH cytochrome c reductase activity in mitochondria isolated from minute amount of human skeletal muscle.
Mitochondria isolated from minute amounts (100-500 mg) of human skeletal muscle displayed a very high rotenone-resistant NADH cytochrome c reductase activity. Moreover, compared to succinate cytochrome c reductase activity, a low rate of rotenone-sensitive NADH cytochrome c reductase activity was measured when using standard procedures to disrupt mitochondrial membranes. Only a drastic osmotic shock in distillated water as a mean to disrupt mitochondrial membrane was found to strongly increase the actual rate of the rotenone-sensitive activity. This was accompanied by a decrease in the rotenone-insensitive activity. Using such a simple procedure, the NADH cytochrome c reductase was found 70-80% inhibited by rotenone and roughly equivalent to 70-85% of the activity of the succinate cytochrome c reductase.
Chretien
D
D
Unité de Recherches sur les Handicap Génétiques de l'Enfant, INSERM U. 12, Hôpital des Enfants Malades, Paris, France.
Bourgeron
T
T
Rötig
A
A
Munnich
A
A
Rustin
P
P
D002458:Cell Fractionation; D005615:Freezing; D006801:Humans; D007700:Kinetics; D008931:Mitochondria, Muscle / Q000201:enzymology* / Q000648:ultrastructure; D009245:NADH Dehydrogenase / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D012402:Rotenone / Q000494:pharmacology*; D014465:Ultrasonics
D012402:Rotenone; D009245:NADH Dehydrogenase