Interaction of malonyl-CoA and related compounds with mitochondria from different rat tissues. Relationship between ligand binding and inhibition of carnitine palmitoyltransferase I.
The sensitivity of carnitine palmitoyltransferase I (CPT I; EC 2.3.1.21) to inhibition by malonyl-CoA and related compounds was examined in isolated mitochondria from liver, heart and skeletal muscle of the rat. In all three tissues the same order of inhibitory potency emerged: malonyl-CoA much greater than succinyl-CoA greater than methylmalonyl-CoA much greater than propionyl-CoA greater than acetyl-CoA. For any given agent, suppression of CPT I activity was much greater in skeletal muscle than in liver, with the heart enzyme having intermediate sensitivity. With skeletal-muscle mitochondria a high-affinity binding site for [14C]malonyl-CoA was readily demonstrable (Kd approx. 25 nM). The ability of other CoA esters to compete with [14C]malonyl-CoA for binding to the membrane paralleled their capacity to inhibit CPT I. Palmitoyl-CoA also competitively inhibited [14C]malonyl-CoA binding, in keeping with its known ability to overcome malonyl-CoA suppression of CPT I. For reasons not yet clear, free CoA displayed anomalous behaviour in that its competition for [14C]malonyl-CoA binding was disproportionately greater than its inhibition of CPT I. Three major conclusions are drawn. First, malonyl-CoA is not the only physiological compound capable of suppressing CPT I, since chemically related compounds, known to exist in cells, also share this property, particularly in tissues where the enzyme shows the greatest sensitivity to malonyl-CoA. Second, malonyl-CoA and its analogues appear to interact with the same site on the mitochondrial membrane, as may palmitoyl-CoA. Third, the degree of site occupancy by inhibitors governs the activity of CPT I.
D000214:Acyl Coenzyme A / Q000494:pharmacology*; D000217:Acyltransferases / Q000037:antagonists & inhibitors*; D000818:Animals; D002334:Carnitine O-Palmitoyltransferase / Q000037:antagonists & inhibitors*; D003065:Coenzyme A / Q000378:metabolism; D066298:In Vitro Techniques; D007700:Kinetics; D008024:Ligands; D008297:Male; D008316:Malonyl Coenzyme A / Q000494:pharmacology*; D008928:Mitochondria / Q000201:enzymology*; D008929:Mitochondria, Heart / Q000201:enzymology; D008930:Mitochondria, Liver / Q000201:enzymology; D008931:Mitochondria, Muscle / Q000201:enzymology; D051381:Rats; D011919:Rats, Inbred Strains; D014018:Tissue Distribution
D000214:Acyl Coenzyme A; D008024:Ligands; D008316:Malonyl Coenzyme A; D000217:Acyltransferases; D002334:Carnitine O-Palmitoyltransferase; D003065:Coenzyme A