A radiochemical assay for mitochondrial acetyl-CoA synthase and octanoyl-CoA synthase from rat heart.
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.
D000106:Acetate-CoA Ligase / Q000378:metabolism*; D000818:Animals; D002250:Carbon Radioisotopes; D003066:Coenzyme A Ligases / Q000378:metabolism*; D005612:Freeze Drying; D007700:Kinetics; D008297:Male; D008929:Mitochondria, Heart / Q000201:enzymology*; D011865:Radioisotope Dilution Technique; D051381:Rats; D011919:Rats, Inbred Strains
D002250:Carbon Radioisotopes; D003066:Coenzyme A Ligases; D000106:Acetate-CoA Ligase; C019805:butyryl-CoA synthetase