Palmitate oxidation by intact preparations of skeletal muscle

1. The palmitate oxidation by intact preparations of rat hemidaphragm, m.soleus and m.flexor digitorum brevis and by teased fibers of human m.pectoralis was studied. 2. The structural and metabolic viability of the in vitro preparations was shown by a low leakage of soluble creatine kinase, a constant rate of palmitate oxidation and only a small stimulatory effect of L-carnitine. 3. With hemidaphragm the palmitate oxidation rate increases with both the palmitate concentration (0-3 mM) and the palmitate/albumin molar ratio (0.5-5.0). 4. The apparent Km for palmitate oxidation was about 1.5 mM at 0.1 and 0.2 mM albumin and about 2.7 mM at 0.4 and 0.6 mM albumin, which correlates with the higher affinity of albumin for palmitate at lower palmitate/albumin molar ratios. 5. After prolonged starvation the apparent Km at 0.4 mM albumin is markedly lower. In whole homogenates of diaphragm the apparent Km at 0.4 mM albumin is only about 370 microM. 6. The calculated maximal oxidation rate was not significantly different for all albumin concentrations examined (23-32 nmol/min per g), did not change after starvation and appears to be of the same order of magnitude as the rate of endogenous fatty acid consumption (30-40 nmol/min per g). 7. Results suggest that substrate availability is a main factor for the oxidation rate of exogenous palmitate by hemidiaphragm in vitro and that the magnitude of the apparent Km is largely dependent upon the degree of label dilution with fatty acids of endogenous origin.