Purification and characterization of tetrahydrofolate.protein complex in bovine liver.
Gel filtration of bovine liver extract on a Sephadex G-200 column resolved three macromolecular fractions with dihydropteridine reductase-dependent cytochrome c reducing activity. One of the active fractions was purified from the extract through the steps of solvent fractionation, chromatography on DEAE-Sephadex, and gel filtration. Biochemical and microbiological analyses showed that the purified complex consists of a Mr = 70,000 protein and tetrahydropteroyldiglutamate. In contrast to the extreme lability of free tetrahydropteridines the complex was quite stable against autooxidation under aerobic conditions.
D000818:Animals; D002417:Cattle; D003579:Cytochrome Reductases / Q000302:isolation & purification / Q000378:metabolism*; D004093:Dihydropteridine Reductase / Q000302:isolation & purification / Q000378:metabolism*; D004355:Drug Stability; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D008970:Molecular Weight; D009097:Multienzyme Complexes / Q000302:isolation & purification / Q000378:metabolism*; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism*; D013329:Structure-Activity Relationship; D013763:Tetrahydrofolates / Q000378:metabolism*
D009097:Multienzyme Complexes; D013763:Tetrahydrofolates; D004093:Dihydropteridine Reductase; D009247:NADH, NADPH Oxidoreductases; D003579:Cytochrome Reductases