title: Purification and characterization of methylenetetrahydrofolate reductase from human cadaver liver.
abstract: Methylenetetrahydrofolate reductase from human cadaver liver was purified to homogeneity. The purified enzyme had a molecular mass of 150 kDa. On SDS-polyacrylamide gel electrophoresis it was dissociated into a single fragment with a molecular mass of 39 kDa. In contrast, fresh lymphocyte enzyme extract showed a major band with a molecular mass of 75 kDa and a minor band of 39 kDa. Fresh liver enzyme was inhibited by S-adenosylmethionine while the purified enzyme from human cadaver liver was not inhibited. These observations suggest that human methylenetetrahydrofolate reductase is composed of two identical subunits of 75 kDa each but is cleaved into a major single band due to autolysis in cadaver liver. The purified cadaver enzyme was a FAD-specific protein. The pH optimum was 6.6 for methylenetetrahydrofolate-NADPH oxidoreductase, 6.5 for methyltetrahydrofolate-menadione oxidoreductase, and 7.2 for NADP-menadione oxidoreductase. The Km values of human liver methylenetetrahydrofolate reductase were 17 microns for NADPH and 38 microns for methyltetrahydrofolate in the reduction of menadione, and 12 microns for NADPH in the reduction of methylenetetrahydrofolate.
authors.affiliation: Department of Pediatrics, Rush Medical College, Chicago, Illinois 60612.
authors.forename: J
authors.initials: J
authors.lastname: Zhou
authors.forename: S S
authors.initials: SS
authors.lastname: Kang
authors.forename: P W
authors.initials: PW
authors.lastname: Wong
authors.forename: B
authors.initials: B
authors.lastname: Fournier
authors.forename: R
authors.initials: R
authors.lastname: Rozen
mesh_terms: D002102:Cadaver; D006801:Humans; D007120:Immunochemistry; D008099:Liver / Q000201:enzymology*; D042965:Methylenetetrahydrofolate Reductase (NADPH2); D008970:Molecular Weight; D000587:Oxidoreductases Acting on CH-NH Group Donors / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D000587:Oxidoreductases Acting on CH-NH Group Donors; D042965:Methylenetetrahydrofolate Reductase (NADPH2)