hexacyanoferrate-iii and Leukemia--Erythroblastic--Acute

The reduction of Fe3+ to Fe2+ has been established to play a critical role in the uptake of iron by many organisms. Recently, a mechanism of iron transport in the absence of transferrin (Tf) was described for the human K562 cell line and a role for ferrireductase activity was implicated in this process as well [Inman, R. S., & Wessling-Resnick, M. (1993) J. Biol. Chem. 268, 8521-8528]. The present report characterizes the extracellular reduction of ferricyanide to ferrocyanide catalyzed by K562 cells. The observation that membrane-impermeant ferricyanide competitively inhibits Tf-independent assimilation of iron from 55Fe-nitriloacetic acid indicates that this ferrireductase activity is indeed coupled to the transport mechanism. From a series of initial rate experiments, the kinetic parameters for cell surface ferrireductase activity, Vmax = 0.102 nmol min-1 (10(6) cells)-1 and Km = 6.13 microM, were determined. Neither the Vmax nor the Km of this reaction is modulated by changes in extra- or intracellular iron levels; thus, similar to Tf-independent transport activity in K562 cells, the ferrireductase activity is not regulated in response to iron levels. Transmembrane oxidoreductase activity is also reportedly involved in the control of cellular growth; however, the K562 cell ferrireductase is unresponsive to insulin and is not inhibited by the antitumor drugs adriamycin, actinomycin D, or cis-platin, observations that fail to support a role for this particular activity in cell regulation. Rather, the K562 cell ferrireductase appears to be tightly coupled to the mechanism of Tf-independent transport as demonstrated by its sensitivity to Cd2+, a specific inhibitor of non-Tf iron uptake by K562 cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Biological Transport; Cadmium; Ferricyanides; Ferrocyanides; FMN Reductase; Gene Expression Regulation, Enzymologic; Humans; Insulin; Iron; Iron Radioisotopes; Kinetics; Leukemia, Erythroblastic, Acute; NADH, NADPH Oxidoreductases; Oxidation-Reduction; Transferrin; Tumor Cells, Cultured

Transmembrane reduction of extracellular oxidants by K562 and U937 leukemic cells was stimulated by catalytic amounts of ascorbate or dehydroascorbate. This stimulation was not due to transport of ascorbate in different redox states in and out of the cells. The membrane redox cycle was strictly dependent on the presence of the cells at every stage, and showed high affinity for ascorbate with simple linear kinetics. Metabolic inhibitors and sulfhydryl reagents inhibited this stimulation. Ascorbate uptake was also dependent on oxidation, but in a very different manner and with much lower affinity for ascorbate. The uptake was non-saturable in the concentration range used. There was some release of ascorbate from the cells, which cannot account for an appreciable part of the reduction of extracellular electron acceptors.

Topics: Ascorbic Acid; Biological Transport; Cell Membrane; Culture Media; Dehydroascorbic Acid; Electron Transport; Ferricyanides; Humans; Kinetics; Leukemia, Erythroblastic, Acute; Lymphoma, Large B-Cell, Diffuse; Oxidation-Reduction; Tumor Cells, Cultured