A ferric reductase activity is found in brush border membrane vesicles isolated from Caco-2 cells.
Brush border membrane vesicles isolated from Caco-2 cells were used to examine whether there is an apical membrane-associated ferric reductase activity in small intestinal enterocytes. A ferric reductase activity which was dependent on NADH or NADPH as reductants was shown. Reduction of Fe(III) was quantified by the formation of a stable Fe(II)/ferrozine complex. The ferric reductase revealed saturation kinetics with a K(m) of 4.12 +/- 0.65 micromol/L and a Vmax of 3.11 +/- 0.043 nmol/(min.mg protein) for NADH. About 25% of the electrons for the NADH-dependent ferric iron reduction were transferred indirectly from the superoxide anion as verified by the superoxide dismutase inhibitable ferric iron reduction rate. However, the main part of Fe(III) reduction occurs directly by catalyzed electron transfer from NADH to ferric iron through (an) enzyme(s) located in the brush border membrane. The ferric reductase activity was inhibited by Pt(II) and especially p-chloromercuribenzoate. Ferricyanide, which is also reduced by the enzyme, is a competitive inhibitor of the Fe(III)/nitrilotriacetate (NTA) complex with a Ki of 43 micromol/L. These results suggest that brush border membranes of enterocytes possess a ferric reductase that reduces ferric to ferrous iron before the iron is transported through the microvillous membrane.
Ekmekcioglu
C
C
University of Vienna, Medical School, Department of Medical Physiology, Austria.
Feyertag
J
J
Marktl
W
W
D018938:Caco-2 Cells / Q000166:cytology / Q000201:enzymology / Q000648:ultrastructure*; D004847:Epithelial Cells; D004848:Epithelium / Q000648:ultrastructure; D038181:FMN Reductase; D005292:Ferricyanides / Q000494:pharmacology; D006801:Humans; D007425:Intracellular Membranes / Q000201:enzymology* / Q000648:ultrastructure; D008871:Microvilli / Q000201:enzymology / Q000648:ultrastructure; D009243:NAD / Q000502:physiology; D009247:NADH, NADPH Oxidoreductases / Q000032:analysis*; D009249:NADP / Q000502:physiology; D013482:Superoxide Dismutase / Q000494:pharmacology
D005292:Ferricyanides; D009243:NAD; C007931:hexacyanoferrate III; D009249:NADP; D013482:Superoxide Dismutase; D038181:FMN Reductase; D009247:NADH, NADPH Oxidoreductases; C023903:ferric citrate iron reductase