title: Synthesis of (6S)-5-formyltetrahydropteroyl-polyglutamates and interconversion to other reduced pteroylpolyglutamate derivatives.
abstract: 5-Formyltetrahydropteroylpolyglutamates can be synthesized and purified directly from dihydropteroylpolyglutamates in a single-step procedure without purification of intermediates and with yields greater than 90%. The procedure involves a coupled enzymatic synthesis of 10-formyltetrahydropteroylpolyglutamates using the enzymes dihydrofolate reductase, serine hydroxymethyltransferase, and C1-tetrahydrofolate synthase with catalytic amounts of NADPH. The 10-formyltetrahydropteroylpolyglutamates are subsequently converted to 5-formyltetrahydropteroylpolyglutamates at 90 degrees C with near quantitative yields. 5-Formyltetrahydropteroylpolyglutamates are the only stable reduced derivatives of tetrahydropteroylpolyglutamates and can be purified and stored indefinitely without decomposition. Additionally, 5-formyltetrahydropteroylpolyglutamates can be readily converted to other derivatives of tetrahydropteroylpolyglutamates with yields greater than 95%. Also described is the synthesis of tetrahydropteroylglutamates labeled at C-11 with either 14C or 13C. Rapid purification procedures for serine hydroxymethyltransferase and C1-tetrahydrofolate synthase from frozen rabbit livers are presented.
authors.affiliation: Department of Biochemistry, Virginia Commonwealth University, Richmond 23298.
authors.forename: P
authors.initials: P
authors.lastname: Stover
authors.forename: V
authors.initials: V
authors.lastname: Schirch
mesh_terms: D000619:Aminohydrolases / Q000302:isolation & purification* / Q000378:metabolism*; D000818:Animals; D002384:Catalysis; D005574:Formate-Tetrahydrofolate Ligase / Q000302:isolation & purification* / Q000378:metabolism*; D012696:Glycine Hydroxymethyltransferase / Q000378:metabolism*; D008099:Liver / Q000201:enzymology*; D008754:Methylenetetrahydrofolate Dehydrogenase (NADP) / Q000302:isolation & purification* / Q000378:metabolism*; D009097:Multienzyme Complexes / Q000302:isolation & purification* / Q000378:metabolism*; D009249:NADP / Q000378:metabolism; D011817:Rabbits
chemical_list: D009097:Multienzyme Complexes; D009249:NADP; C021836:formyl-methenyl-methylenetetrahydrofolate synthetase; D008754:Methylenetetrahydrofolate Dehydrogenase (NADP); D012696:Glycine Hydroxymethyltransferase; D000619:Aminohydrolases; D005574:Formate-Tetrahydrofolate Ligase