Cytochemical demonstration of 5-formyl tetrahydrofolate cyclodehydrase and 5,10-methenyl tetrahydrofolate cyclohydrolase activity.
The enzyme 5-formyl tetrahydrofolate cyclodehydrase plays an important role in the conversion of 5-formyl tetrahydrofolate to 5,10-methenyl tetrahydrofolate. A second enzyme, cyclohydrolase, converts 5,10-methenyl tetrahydrofolate to 10-formyl tetrahydrofolate. These folate derivatives play a significant part in the biosynthesis of purines. A method has been devised for the cytochemical demonstration of 5-formyl tetrahydrofolate cyclodehydrase and 5,10-methenyl tetrahydrofolate cyclohydrolase activity which uses 5-formyl tetrahydrofolate or 5,10-methenyl tetrahydrofolate as substrate respectively, blocking possible interferences by other enzymes, and allows the nonenzymatic reduction of nitro-blue tetrazolium by 5,10-methenyl tetrahydrofolate formed by the action of the cyclodehydrase on the substrate 5-formyl tetrahydrofolate, and by 10-formyl tetrahydrofolate formed by the action of cyclohydrolase on the substrate 5,10-methenyl tetrahydrofolate, thus revealing intracellular sites of enzyme activity. The methods appear to show only intracellular localization of the blue formazan deposits of reduced tetrazolium. The distribution of positivity in cells of human blood and bone marrow is described.
D000619:Aminohydrolases / Q000037:antagonists & inhibitors / Q000097:blood / Q000378:metabolism*; D001853:Bone Marrow / Q000201:enzymology; D001854:Bone Marrow Cells; D019731:Carbon-Nitrogen Ligases; D002729:Chloromercuribenzoates / Q000494:pharmacology; D005575:Formyltetrahydrofolates / Q000097:blood / Q000378:metabolism*; D006651:Histocytochemistry; D006801:Humans; D002955:Leucovorin; D008025:Ligases / Q000037:antagonists & inhibitors / Q000097:blood / Q000378:metabolism*; D008727:Methotrexate / Q000494:pharmacology; D013763:Tetrahydrofolates / Q000378:metabolism*
D002729:Chloromercuribenzoates; D005575:Formyltetrahydrofolates; D013763:Tetrahydrofolates; D000619:Aminohydrolases; D008025:Ligases; D019731:Carbon-Nitrogen Ligases; C021557:5,10-methenyltetrahydrofolate synthetase; D002955:Leucovorin; D008727:Methotrexate