Evaluation of Folylpolyglutamates by Electrophoretic Separation of Fluorodeoxyuridylate-Thymidylate Synthase-Methylenetetrahydrofolate Complexes
The chain length of specific reduced folylpolyglutamates has been estimated by incorporation of the 5,10-Tnethylenetetrahydrofolate form of the cofactor into a stable ternary complex with L. casei thymidylate synthase and tritiated fluorodeoxyuridylate followed by electrophoretic separation based on charge differences in complexed polyglutamates. The method is also applicable to tetrahydrofolate polyglutamates after conversion to the active cofactor form by introduction of formaldehyde. The method can be used to analyze less than one pmole of folylpolyglut-amate and can be applied to evaluation of tissue polyglutamates or to monitor relevant enzyme catalyzed reactions.
KeywordsTernary Complex Electrophoretic Separation Glutamate Residue Longe Chain Length Folate Pool
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