title: Diferric transferrin reduction by K562 cells. A critical study.
abstract: This paper critically examines the redox activity of K562 cells (chronic myelogenous leukemia cells) and normal peripheral blood lymphocytes (PBL). Ferricyanide reduction, diferric transferrin reduction, and ferric ion reduction were measured spectrophotometrically by following the time-dependent changes of absorbance difference characteristic for ferricyanide disappearance and for the formation of ferrous ion:chelator complexes. Bathophenanthroline disulfonate (BPS) and ferrozine (FZ) were used to detect the appearance of ferrous ions in the reaction mixtures when diferric transferrin or ferric reduction was studied. Special attention was devoted to the analysis of time-dependent absorbance changes in the presence and absence of cells under different assay conditions. It was observed and concluded that: (i) FZ was far less sensitive and more sluggish than BPS for detecting ferrous ions at concentrations commonly used for BPS; (ii) FZ, at concentrations of at least 10-times the commonly used BPS concentrations, seemed to verify the results obtained with BPS; (iii) ferricyanide reduction, diferric transferrin reduction and ferric ion reduction by both K562 cells and peripheral blood lymphocytes did not differ significantly; and (iv) earlier values published for the redox activities of different cells might be overestimated, partly because of the observation published in 1988 that diferric transferrin might have loosely bound extra iron which is easily reduced. It is suggested that the specific diferric transferrin reduction by cells might be considered as a consequence of (i) changing the steady-state equilibrium in the diferric transferrin-containing solution by addition of ferrous ion chelators which effectively raised the redox potential of the iron bound in holotransferrin, and (ii) changing the steady-state equilibrium by addition of cells which would introduce, via their large and mostly negatively charged plasma membrane surface, a new phase which would favor release and reduction of the iron in diferric transferrin by a ferric ion oxidoreductase. The reduction of ferricyanide is also much slower than activities reported for other cells which may indicate reduced plasma membrane redox activity in these cells.
authors.affiliation: Center for Reproduction and Transplantation Immunology, Methodist Hospital of Indiana, Indianapolis 46202.
authors.forename: A
authors.initials: A
authors.lastname: Bérczi
authors.forename: J A
authors.initials: JA
authors.lastname: Sizensky
authors.forename: F L
authors.initials: FL
authors.lastname: Crane
authors.forename: W P
authors.initials: WP
authors.lastname: Faulk
mesh_terms: D005292:Ferricyanides / Q000378:metabolism; D005297:Ferrozine; D006801:Humans; D007700:Kinetics; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000378:metabolism*; D008214:Lymphocytes / Q000378:metabolism; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism; D010084:Oxidation-Reduction; D010618:Phenanthrolines; D015203:Reproducibility of Results; D013053:Spectrophotometry; D014168:Transferrin / Q000378:metabolism*; D014407:Tumor Cells, Cultured
chemical_list: D005292:Ferricyanides; D010618:Phenanthrolines; D014168:Transferrin; C028283:diferric transferrin; D005297:Ferrozine; C017049:bathophenanthroline disulfonic acid; D009247:NADH, NADPH Oxidoreductases; C050129:diferric transferrin reductase; C028033:potassium ferricyanide