title: Monocyte-macrophage ferric reductase activity is inhibited by iron and stimulated by cellular differentiation.
abstract: The enzyme ferric reductase catalyses the reduction of Fe(III) as a prerequisite to its transportation across the cell membrane. Duodenal mucosal biopsies from iron overloaded patients with genetic haemochromatosis (GH) have increased ferric reductase activity and iron absorption compared with controls, yet the GH mucosa is iron deficient. A similar GH-related iron deficiency is also seen in macrophages. The aim of this study was to investigate whether macrophage ferric reductase activity is altered in GH, and to determine ferric reductase activity in monocytes and differentiated macrophages. The erythroleukaemic K562 cell line was studied as a clonal reference cell line. The basal K562 ferric reductase activity is characteristic of a membrane bound enzyme, being both temperature and protease sensitive. Ferric reductase activity was also demonstrated in human leucocyte, monocyte and macrophage preparations. Assays of K562 and macrophage cell supernatants confirmed that the ferric reductase activity was not due to a secreted factor. Assay of ferric reductase in normalized-iron and iron-enriched (100 microM ferric citrate) conditions showed no significant difference between Cys282Tyr (Cys282-->Tyr) homozygous GH macrophages and Cys282-Tyr negative control activities (P>0.05). However, a 900% increase in ferric reductase activity was observed during monocyte to macrophage differentiation (P<0.05), possibly reflecting the co-ordinate up-regulation of iron metabolism in these cells. The demonstration of approx. 25% activity after macrophage differentiation at high free-iron concentrations compared with 'normalized' iron is consistent with repression of human ferric reductase activity by iron. The identification of the human ferric reductase gene and its protein will ultimately provide insight into its regulation and role in mammalian iron metabolism.
authors.affiliation: Department of Medicine, Royal Free and University College Medical School, Royal Free Campus, Rowland Hill Street, London NW3 2PF, U.K.
authors.forename: J
authors.initials: J
authors.lastname: Partridge
authors.forename: D F
authors.initials: DF
authors.lastname: Wallace
authors.forename: K B
authors.initials: KB
authors.lastname: Raja
authors.forename: J S
authors.initials: JS
authors.lastname: Dooley
authors.forename: A P
authors.initials: AP
authors.lastname: Walker
mesh_terms: D002454:Cell Differentiation; D002478:Cells, Cultured; D038181:FMN Reductase; D006432:Hemochromatosis / Q000201:enzymology; D006801:Humans; D007501:Iron / Q000494:pharmacology*; D004915:Leukemia, Erythroblastic, Acute / Q000201:enzymology; D008264:Macrophages / Q000201:enzymology*; D008297:Male; D009000:Monocytes / Q000201:enzymology*; D009247:NADH, NADPH Oxidoreductases / Q000037:antagonists & inhibitors*
chemical_list: D007501:Iron; D038181:FMN Reductase; D009247:NADH, NADPH Oxidoreductases; C023903:ferric citrate iron reductase