Identification of 2-tetradecylglycidyl coenzyme A as the active form of methyl 2-tetradecylglycidate (methyl palmoxirate) and its characterization as an irreversible, active site-directed inhibitor of carnitine palmitoyltransferase A in isolated rat liver mitochondria.
Methyl-2-tetradecylglycidic acid (MeTDGA) has been hypothesized to inhibit fatty acid oxidation by irreversible, active site-directed inactivation of carnitine palmitoyltransferase A after being converted to TDGA-CoA. Using synthetic TDGA-CoA, this hypothesis has been confirmed. Assessing enzyme inhibition in an isolated rat liver mitochondrial system, TDGA-CoA (synthetic or enzyme prepared) was more potent than TDGA or MeTDGA and retained activity in the absence of CoA or Mg2+-ATP. It inhibited palmitoyl-CoA but not palmitoyl carnitine oxidation. Enzyme inactivation was exponential, stereospecific, and fast (t0.5 = 38.5 s with 100 nM (R)-TDGA-CoA). TDGA-CoA was identified as a complexing type irreversible inhibitor (Ki approximately 0.27 microM) by the double reciprocal relationship between the pseudo-first order inactivation rate and its concentration, by the inverse dependence of the second order rate constant on its concentration, and by the independence of the first order rate from the enzyme concentration. Palmitoyl-CoA, CoA, and malonyl-CoA protected the enzyme, while L-carnitine and palmitoyl-L-carnitine were without effect. [3-14C] TDGA-CoA labeled a protein, Mr = 90,000, with a time course which paralleled that of enzyme inhibition; maximum specific binding was 16 pmol/mg of mitochondrial protein.
D000214:Acyl Coenzyme A / Q000031:analogs & derivatives*; D000217:Acyltransferases / Q000037:antagonists & inhibitors*; D000818:Animals; D001384:Azetidines; D001665:Binding Sites; D002334:Carnitine O-Palmitoyltransferase / Q000037:antagonists & inhibitors*; D004852:Epoxy Compounds / Q000494:pharmacology*; D004988:Ethers, Cyclic / Q000494:pharmacology*; D005227:Fatty Acids / Q000378:metabolism; D007700:Kinetics; D008930:Mitochondria, Liver / Q000201:enzymology*; D010084:Oxidation-Reduction; D010171:Palmitoyl Coenzyme A / Q000031:analogs & derivatives* / Q000494:pharmacology; D011422:Propionates / Q000494:pharmacology*; D051381:Rats; D011919:Rats, Inbred Strains; D013237:Stereoisomerism
D000214:Acyl Coenzyme A; D001384:Azetidines; D004852:Epoxy Compounds; D004988:Ethers, Cyclic; D005227:Fatty Acids; D011422:Propionates; D010171:Palmitoyl Coenzyme A; C042420:2-tetradecylglycidyl-coenzyme A; D000217:Acyltransferases; D002334:Carnitine O-Palmitoyltransferase; C017522:methyl 2-tetradecylglycidate