Purification of and mechanism studies on citrate synthase. Use of biospecific adsorption-elution techniques.

Citrate synthases from animal tissues were found to bind to Sepharose-"ATP." A pure preparation of citrate synthase was obtained from a crude fraction of rat heart by the specific elution of the enzyme from the Sepharose-"ATP" with the dead end complex-forming substrates, oxalacetate and CoA. The proposed mechanisms of citrate synthase, obtained from steady state kinetics, were examined in light of the elution pattern of the enzyme obtained using combinations of substrates and substrate analogs.