One-step synthesis of radioactive acyl-CoA and acylcarnitines using rat liver mitochondrial outer membrane as enzyme source.
Rat liver mitochondrial outer membrane enriched preparations have proven to be a convenient enzyme source for synthesizing coenzyme A (CoA) and carnitine esters of radioactive fatty acids. These membranes are simple to isolate and they retain acyl-CoA ligase and carnitine palmitoyltransferase activities well upon storage. Enzyme purification is not required. A novel aspect of the present procedure is that the same enzymatic incubation step allows both the acyl-CoA and the acylcarnitine esters to be obtained simultaneously when carnitine is present, but produces acyl-CoA ester only when carnitine is not included. Under the conditions described, the conversion of [1-14C]octanoic acid to the respective esters was about 95%; the corresponding figure for [1-14C]palmitic acids was over 70%. The procedure seems suitable for synthesizing the labeled CoA and carnitine esters from a variety of radioactive fatty acids.
Bhuiyan
A K
AK
Laboratory of Intermediary Metabolism, Clinical Research Institute of Montreal, Quebec, Canada.
Pande
S V
SV
D000214:Acyl Coenzyme A / Q000138:chemical synthesis*; D000215:Acylation; D000818:Animals; D002331:Carnitine / Q000031:analogs & derivatives* / Q000138:chemical synthesis; D007425:Intracellular Membranes / Q000201:enzymology*; D008930:Mitochondria, Liver / Q000201:enzymology*; D051381:Rats
D000214:Acyl Coenzyme A; D002331:Carnitine