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A radiochemical assay for mitochondrial acetyl-CoA synthase and octanoyl-CoA synthase from rat heart |
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| Authors: | K C Man J T Brosnan |
| Affiliation: | 1. Division of Genetic and Genomic Medicine, Department of Pediatrics, University of Pittsburgh, Pittsburgh, PA, USA;2. Department of Human Genetics, University of Pittsburgh, Pittsburgh, PA, USA;3. Moderna Therapeutics, Inc., Rare Diseases, 200 Technology Square, Cambridge, MA, USA |
| Abstract: | A simple radiochemical assay is described for measuring the activity of rat heart mitochondrial acetyl-CoA synthase (EC 6.2.1.1) and octanoyl-CoA synthase (EC 6.2.1.2) using labelled acetate and octanoate. Separation of 14C-labelled reactant from its reaction products was achieved by lyophilization. Enzyme activity was determined by the measurement of incorporation of 14C-labelled short and medium-chain fatty acid into their CoA-derivatives. The method is applicable to the assay of other enzymes where products and substrate may be separated on the basis of the volatilization of one of them during lyophilization. |
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