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  • Title: A coupled radiometric assay for dihydrofolate reductase based on the oxidative decarboxylation of D-6-phospho-[1-14C]gluconate.
    Author: Rathod PK, Reyes P.
    Journal: Anal Biochem; 1983 Sep; 133(2):425-9. PubMed ID: 6638503.
    Abstract:
    The synthesis of D-6-phospho-[1-14C]gluconate and the utilization of this compound in a novel assay procedure for dihydrofolate reductase is described. This new assay method couples reductase-dependent NADP+ production to the enzymatic and NADP+-dependent decarboxylation of D-6-phospho-[1-14C]gluconate. By several criteria it is demonstrated that [14C]CO2 release is directly proportional to dihydrofolate reductase activity. This coupled radiometric assay for dihydrofolate reductase is more sensitive than the commonly used spectrophotometric assay and offers a number of advantages over earlier radiometric methods.
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