Effect of Fe3+ on Na,K-ATPase: Unexpected activation of ATP hydrolysis

Iron is a key element in cell function; however, its excess in iron overload conditions can be harmful through the generation of reactive oxygen species (ROS) and cell oxidative stress. Activity of Na, K-ATPase has been shown to be implicated in cellular iron uptake and iron modulates the Na, K-ATPase function from different tissues. In this study, we determined the effect of iron overload on Na, K-ATPase activity and established the role that isoforms and conformational states of this enzyme has on this effect. Total blood and membrane preparations from erythrocytes (ghost cells), as well as pig kidney and rat brain cortex, and enterocytes cells (Caco-2) were used. In E1-related subconformations, an enzyme activation effect by iron was observed, and in the E2-related subconformations enzyme inhibition was observed. The enzyme's kinetic parameters were significantly changed only in the Na+ curve in ghost cells. In contrast to Na, K-ATPase alpha 2 and alpha 3 isoforms, activation was not observed for the alpha 1 isoform. In Caco-2 cells, which only contain Na, K-ATPase alpha 1 isoform, the FeCl(3 )increased the intracellular storage of iron, catalase activity, the production of H2O2 and the expression levels of the alpha 1 isoform. In contrast, iron did not affect lipid peroxidation, GSH content, superoxide dismutase and Na, K-ATPase activities. These results suggest that iron itself modulates Na, K-ATPase and that one or more E1-related subconformations seems to be determinant for the sensitivity of iron modulation through a mechanism in which the involvement of the Na, K-ATPase alpha 3 isoform needs to be further investigated.