I. Ghadiminejad, D. Saggerson
May 8, 1991
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0
Influential Citations
12
Citations
Quality indicators
Journal
Biochimica et biophysica acta
Abstract
Sodium cholate was used as an anionic detergent to discriminate the two components of liver overt carnitine palmitoyltransferase (CPT1); namely a catalytic entity and a regulatory component that bound malonyl-CoA. Cholate solubilized approx. 40% of the malonyl-CoA binding entity from mitochondrial outer membranes without appreciable solubilization of CPT1 activity. Cholate did not interfere with binding of [14C]malonyl-CoA to outer membranes or to crude total mitochondrial membrane fractions. By contrast, the non-ionic detergent Tween-20 was ineffective in solubilizing the malonyl-CoA binding entity and also substantially interfered with the binding of [14C]malonyl-CoA. Both detergents appeared to cause total disengagement of the malonyl-CoA binding entity from the catalytic entity of CPT1 only when some inner membrane material was present. 'Reconstitution' experiments were performed in which a malonyl-CoA sensitivity conferring factor in cholate extracts from outer membranes was associated with CPT derived from inner membranes (CPT2). The IC50 for inhibition of CPT2 by malonyl-CoA in this artificial system was similar to that observed with CPT1 in situ in outer membranes. Extracts containing malonyl-CoA sensitivity conferring factor derived from outer membranes of fed or 48 h fasted rats were associated with CPT2 derived from fed rats. The outer membrane extracts from fasted animals conferred a lower maximum responsiveness to malonyl-CoA, but appeared to have a higher affinity for CPT2 than the extracts from fed rats. These results suggest that physiological state can alter the intrinsic properties of the malonyl-CoA sensitivity confering factor.