A novel iron uptake mechanism mediated by GPI‐anchored human p97

The established process for iron uptake into mammalian cells involves transferrin and its receptor. Here, the role of the glycosyl‐phosphatidylinositol (GPI)‐linked transferrin homologue, melanotransferrin or p97, was studied using CHO cell lines defective in the transferrin receptor (TR) and transfected with human TR and/or human p97. The presence of p97 doubled the iron uptake, which could be explained by the binding of one atom of iron to one molecule of p97. The internalization of iron was shown to be temperature sensitive and saturated at a media iron concentration of 2.5 micrograms/ml with a Vmax of 0.1 pmol Fe/10(6) cell/min and a Km of 2.58 microM for p97. Treatment of the cells with either phosphatidylinositol‐phospholipase C or monoclonal antibodies against p97 resulted in over a 50% reduction and a 47% increase in the iron uptake respectively. These data identify p97 as a unique cell surface GPI‐anchored, iron binding protein involved in the transferrin‐independent uptake of iron in mammals.