Regulation of the biosynthesis of CoA at the level of pantothenate kinase.
1. Pantothenate kinase, which is present in cytosol, was studied in preparations from livers of rats fed normal or clofibrate-enriched diets. Effects of CoA, dephospho-CoA and different acyl-CoA derivatives on this enzyme activity were examined in vitro. 2. With partially purified pantothenate kinase or crude particle-free supernatant from the liver of normal or clofibrate-treated rats, Km for pantothenic acid was 0.016 mmol/l at the pH optimum 6.1. 3. Acetyl-CoA, propionyl-CoA, malonyl-CoA and other short-chain acyl-CoA derivatives were strong inhibitors of pantothenate kinase, with Ki in the range 0.001-0.003 mmol/l. The mechanism of inhibition appeared to be of an uncompetitive type. 4. Free CoA has been held to be the main regulator of pantothenate kinase. We found, however, that free CoASH, dephospho-CoA and long-chain acyl-CoA (with Ki 0.003-0.08 mmol/l) were less efficient inhibitors than acetyl-CoA. 5. With pantothenate kinase from clofibrate-treated animals, all inhibitors were less potent. This was most pronounced when the enzyme was assayed in a crude supernatant fraction, possibly because the inhibitors were degraded and/or protein bound. Such a reduction of normal inhibition may contribute to the increased biosynthesis of CoA previously observed during clofibrate treatment. 6. Fasting or diabetes leads to an increase of long-chain acyl-CoA and total CoA in the liver. The increase of CoA has been explained by increased acylation of CoA, and thereby reduced feed-back inhibition by free CoASH at the pantothenate kinase level. We propose another explanation. In these metabolic states, the cytosolic pool of acetyl-CoA is decreased. Since pantothenate kinase is present only in the cytosol, its activity will be released and the biosynthesis of CoA will increase. 7. Acetyl-CoA is probably a more important physiological regulator of pantothenate kinase activity than is free CoASH.
D000105:Acetyl Coenzyme A / Q000502:physiology*; D000214:Acyl Coenzyme A / Q000494:pharmacology; D000818:Animals; D002845:Chromatography / Q000379:methods; D003065:Coenzyme A / Q000096:biosynthesis*; D003600:Cytosol / Q000201:enzymology; D008099:Liver / Q000201:enzymology*; D008297:Male; D010764:Phosphorylase Kinase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains
D000214:Acyl Coenzyme A; D000105:Acetyl Coenzyme A; D010764:Phosphorylase Kinase; D003065:Coenzyme A