Chemical synthesis of folylpolyglutamates, their reduction to tetrahydro derivatives, and their activity with yeast C1-THF synthase.
Pteroyldi- through -pentaglutamic acids were synthesized through the use of the carbodiimide method, rather than the anhydride method or solid phase synthesis. The compounds were converted to the tetrahydro derivatives by enzymatic reduction. The Km values of the folate coenzymes for yeast C1-THF synthase were determined. The determination of the values for the formyltetrahydrofolate synthetase activity of the multifunctional enzyme was possible through the use of newly devised assay based on the fluorescence properties of the pteridine derivatives. The value for the tetraglutamyl coenzyme derivative was approximately 1000-fold lower than that of the monoglutamyl coenzyme, tetrahydrofolate.
D000619:Aminohydrolases / Q000378:metabolism; D000818:Animals; D002417:Cattle; D005492:Folic Acid / Q000031:analogs & derivatives*; D005574:Formate-Tetrahydrofolate Ligase / Q000378:metabolism; D007202:Indicators and Reagents; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D008754:Methylenetetrahydrofolate Dehydrogenase (NADP) / Q000378:metabolism; D009097:Multienzyme Complexes / Q000378:metabolism*; D009154:Mutation; D010084:Oxidation-Reduction; D011485:Protein Binding; D011624:Pteroylpolyglutamic Acids / Q000138:chemical synthesis* / Q000494:pharmacology; D051381:Rats; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D012756:Sheep; D013045:Species Specificity; D013552:Swine
D007202:Indicators and Reagents; D009097:Multienzyme Complexes; D011624:Pteroylpolyglutamic Acids; C021836:formyl-methenyl-methylenetetrahydrofolate synthetase; D005492:Folic Acid; D008754:Methylenetetrahydrofolate Dehydrogenase (NADP); D000619:Aminohydrolases; D005574:Formate-Tetrahydrofolate Ligase