Stoichiometric hydrolysis of long chain acyl-CoA and measurement of the CoA formed with an enzymatic cycling method

Abstract

A new method for measuring long chain acyl-CoA is presented in this paper. It consists of two main procedures: stoichiometric hydrolysis of long chain acyl-CoA with fatty acid synthetase and measurement of the CoA formed with an enzymatic cycling system containing citrate synthase, carnitine acetyl transferase, and malate dehydrogenase. The concentration of long chain acyl-CoA was determined in freeze-clamped liver and “freeze-blown” brain. The values obtained for liver are similar to those previously reported. The values obtained for brain are 11.3 ± 0.8 nmole/g wet wt in 48 hr starved rats and 11.2 ± 0.6 nmole/g wet wt in fed rats.