title: Kinetic mechanism of the cytosolic malic enzyme from human breast cancer cell line.
abstract: The kinetic mechanism of the cytosolic NADP(+)-dependent malic enzyme from cultured human breast cancer cell line was studied by steady-state kinetics. In the direction of oxidative decarboxylation, the initial-velocity and product-inhibition studies indicate that the enzyme reaction follows a sequential ordered Bi-Ter kinetic mechanism with NADP+ as the leading substrate followed by L-malate. The products are released in the order of CO2, pyruvate, and NADPH. The enzyme is unstable at high salt concentration and elevated temperature. However, it is stable for at least 20 min under the assay conditions. Tartronate (2-hydroxymalonate) was found to be a noncompetitive inhibitor for the enzyme with respect to L-malate. The kinetic mechanism of the cytosolic tumor malic enzyme is similar to that for the pigeon liver cytosolic malic enzyme but different from those for the mitochondrial enzyme from various sources.
authors.affiliation: Department of Biochemistry, National Defense Medical Center, Taipei, Taiwan, Republic of China.
authors.forename: G G
authors.initials: GG
authors.lastname: Chang
authors.forename: T M
authors.initials: TM
authors.lastname: Huang
authors.forename: J K
authors.initials: JK
authors.lastname: Wang
authors.forename: H J
authors.initials: HJ
authors.lastname: Lee
authors.forename: W Y
authors.initials: WY
authors.lastname: Chou
authors.forename: C L
authors.initials: CL
authors.lastname: Meng
mesh_terms: D000818:Animals; D001943:Breast Neoplasms / Q000201:enzymology*; D002245:Carbon Dioxide / Q000378:metabolism; D010856:Columbidae; D003600:Cytosol / Q000201:enzymology*; D004795:Enzyme Stability; D006801:Humans; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D008291:Malate Dehydrogenase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D008293:Malates / Q000378:metabolism; D009249:NADP / Q000378:metabolism; D011773:Pyruvates / Q000378:metabolism; D019289:Pyruvic Acid; D013646:Tartronates / Q000494:pharmacology; D014407:Tumor Cells, Cultured
chemical_list: D008293:Malates; D011773:Pyruvates; D013646:Tartronates; D002245:Carbon Dioxide; C032973:tartronic acid; D009249:NADP; C030298:malic acid; D019289:Pyruvic Acid; D008291:Malate Dehydrogenase