Iron transport across brush-border membranes from normal and iron-deficient mouse upper small intestine.

We have studied Fe(III)-citrate and Fe(II)-ascorbate uptake by purified intestinal brush-border membrane vesicles from normal (iron-replete) and iron-deficient mice. In iron-replete mice using a final Fe(III) concentration of 1.43 microM, 25-30 pmol of Fe(III)/mg of protein were bound to the membranes versus 65-70 pmol in iron-deficient mice. Fe(II) uptake in normal mice using a final Fe(II) concentration of 1.79 microM was 1600-1800 pmol/mg of protein versus 3600-4000 pmol in iron-deficient mice. Evidence that Fe(II) was transported into the vesicles by a membrane carrier-mediated process was obtained by observing saturation kinetics under conditions of isotope exchange at equilibrium in mice rendered iron-deficient, but not in iron-replete mice. Eighty per cent of the transported Fe(II) could be removed by strong chelating agents. The remainder was exchangeable with Fe(II) in the medium when measured under equilibrium conditions. We can explain these results by the following model; iron uptake appears to be a 2-fold process. The first step is the transport of Fe(II) across the membrane by a carrier-mediated process which is biologically regulated. The second step is the subsequent binding of iron on the inside of the membrane. The number of binding sites is also regulated by the iron status of the mouse. The membrane binding affinity for Fe(II) appears to be weaker than that for dithiothreitol but stronger than for ascorbate.