Abstract
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 simulataneously 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.
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Abbreviations
- CoA:
-
coenzyme A
- CPTo :
-
outer carnitine palmitoyl-transferase
- EDTA:
-
ethylenediaminetetraacetic acid
- HEPES:
-
N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid
- OMV:
-
outer membrane enriched vesicles
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A preliminary account of this work has been published (ref. 1).
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Jalaluddin Bhuiyan, A.K.M., Pande, S.V. One-step synthesis of radioactive acyl-CoA and acylcarnitines using rat liver mitochondrial outer membrane as enzyme source. Lipids 27, 392–395 (1992). https://doi.org/10.1007/BF02536155
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DOI: https://doi.org/10.1007/BF02536155