After the determination of basal respiration rate measured at 22°C with R solution plus 1 mM pyruvate and 4 mM malate as mitochondrial substrates (V0, nonphosphorylating rate), fibers were exposed to an increasing concentration of ADP to determine the apparent Michaelis-Menten constant (Km) for ADP and maximal mitochondrial respiration [at saturating concentration of ADP (2 mM) Vmax, phosphorylating rate]. The ratio of Vmax/V0 represented the degree of coupling between oxidation and phosphorylation of ADP for pyruvate.

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