Electrophoretic identification of poly-gamma-glutamate chain-lengths of 5,10-methylenetetrahydrofolate using thymidylate synthetase complexes.
A method has been developed to characterize the poly-gamma-glutamates of 5,10-methylenetetrahydrofolate. Incorporation of 5,10-methylenetetrahydrofolates into a ternary complex with L. casei thymidylate synthetase and 5-fluoro-2-deoxy[3H]uridylate stabilizes the reduced folate against oxidation, loss of the one carbon moiety, and poly-gamma-glutamate degradation. The covalent ternary complexes, containing 5,10-methylenetetrahydrofolate polyglutamates, were resolved electrophoretically. Electrophoretic mobility was shown to be a linear function of polyglutamate chain-length. The method can potentially be applied to analysis of chemically prepared folate polyglutamates, the monitoring of enzyme-mediated interconversions of polyglutamates and characterization of tissue extract polyglutamates.
D055598:Chemical Phenomena; D002621:Chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D005492:Folic Acid / Q000031:analogs & derivatives*; D008780:Methyltransferases; D011624:Pteroylpolyglutamic Acids / Q000302:isolation & purification*; D013763:Tetrahydrofolates; D013940:Thymidylate Synthase
D011624:Pteroylpolyglutamic Acids; D013763:Tetrahydrofolates; C013123:5,10-methylenetetrahydrofolic acid; D005492:Folic Acid; D008780:Methyltransferases; D013940:Thymidylate Synthase