Radioisotopic assay of picomolar amounts of coenzyme A.
A two-step method of determining reduced coenzyme A (CoASH) concentrations in tissue or cell extracts is described. In the first step, CoASH is reacted with acetylphosphate in a reaction catalyzed by phosphotransacetylase to yield acetyl-CoA. Acetyl-CoA is then condensed with [14C]oxaloacetate by citrate synthase to give [14C]citrate. This method allows the measurement of 10-200 pmol of CoASH. By omitting the phosphotransacetylase step, measurement of the same amount of acetyl-CoA is possible.
D000105:Acetyl Coenzyme A / Q000302:isolation & purification; D000818:Animals; D002384:Catalysis; D003065:Coenzyme A / Q000302:isolation & purification*; D007553:Isotope Labeling; D008832:Microchemistry; D008930:Mitochondria, Liver / Q000201:enzymology; D010084:Oxidation-Reduction; D051381:Rats
D000105:Acetyl Coenzyme A; D003065:Coenzyme A