title: Fe(II) oxidation and Fe(III) incorporation by the M(r) 66,000 microsomal iron protein that stimulates NADPH oxidation.
abstract: In a previous study (Minotti, G., and Ikeda-Saito, M. (1991) J. Biol. Chem. 266, 20011-20017) we demonstrated the existence of a M(r) 66,000 microsomal iron protein (MIP) which stimulates NADPH oxidation by shunting electrons from NADPH-cytochrome P-450 reducase to its bound Fe(III). In the present study, purified MIP was depleted of iron and the apoMIP was examined for its ability to incorporate Fe(III) upon an incubation with Fe(II). It was found that apoMIP had an oxygen-dependent ferroxidase activity coupled with the incorporation of Fe(III). The reconstituted MIP exhibited a Fe(III) content and an NADPH oxidation activity similar to those of native MIP. However, the reconstitution of MIP from apoMIP and Fe(II) had to be performed in the presence of detergents to prevent the formation of protein aggregates and the oxidative incorporation of an iron which could not react with NADPH-cytochrome P-450 reductase. This redox inactive iron was probably bound nonspecifically to artifactual sites formed by the protein aggregates.
authors.affiliation: Institute of General Pathology, Catholic University School of Medicine, Rome, Italy.
authors.forename: G
authors.initials: G
authors.lastname: Minotti
authors.forename: M
authors.initials: M
authors.lastname: Ikeda-Saito
mesh_terms: D000818:Animals; D002417:Cattle; D002570:Ceruloplasmin / Q000378:metabolism; D002853:Chromatography, Liquid; D007501:Iron / Q000378:metabolism*; D008667:Metalloproteins / Q000302:isolation & purification / Q000378:metabolism*; D008861:Microsomes / Q000378:metabolism*; D009206:Myocardium / Q000378:metabolism*; D009249:NADP / Q000378:metabolism*; D009251:NADPH-Ferrihemoprotein Reductase / Q000378:metabolism; D019159:Nonheme Iron Proteins; D010084:Oxidation-Reduction
chemical_list: D008667:Metalloproteins; D019159:Nonheme Iron Proteins; D009249:NADP; D007501:Iron; D002570:Ceruloplasmin; D009251:NADPH-Ferrihemoprotein Reductase