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Clinical Chemistry 19: 1106-1111, 1973;
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Clinical Chemistry, Vol 19, 1106-1111, Copyright © 1973 by the American Association for Clinical Chemistry

Radiometric Assay of Citrate Condensing Enzyme (Citrate Synthetase) in Liver Tissue and Mitochondria

Padmakar K. Dixit 1 and Raul Cuestas 1

1 Department of Anatomy, 272 Jackson Hall, University of Minnesota School of Medicine, Minneapolis, Minn. 55455.

Citrate synthetase activity was determined in samples of commercially available citrate condensing enzyme. We measured the amount of radioactive citrate formed from the condensation of carbon-14 labeled acetyl CoA and oxaloacetate. The labeled citrate was oxidized by oxidative bromination; 14CO2 evolved was collected and represented the amount of citrate synthesized. Enzyme activity as low as 10-4 µmol/min was measured reproducibly with this procedure. Results obtained with this assay compared closely with those of a widely used spectrophotometric assay in which Ellman’s SH-coupling reagent [5,5-dithio-bis-(2-nitrobenzoate)] is coupled with CoA released during the reaction. Accuracy of labeled citrate determination in the presence of labeled acetate or pyruvate was excellent. The method can be used to determine citrate synthetase activity in skeletal muscle, liver, and isolated liver mitochondria, or to study the fate of labeled citrate injected into animals.


Key Words: diagnosis of diabetes, rickets • rats • citrate metabolism • acetyl CoA

Submitted on May 2, 1973
Accepted on July 9, 1973






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Copyright © 1973 by the American Association for Clinical Chemistry.