A single-step enzyme assay using [14C]palmitic acid and bacterial acyl-coenzyme A synthetase (EC 220.127.116.11) is described for the determination of reduced coenzyme A (CoASH) levels in liver samples. Use of this technique provides a rapid and accurate determination of CoASH in the range 1-250 pmol. Application of the method to the quantitation of CoASH in samples of human liver tissue and rat liver homogenate, isolated hepatocytes, and mitochondria is described.
- coenzyme A
- human liver
- liquid scintillation counting