Analyses for binding of the transferrin family of proteins to the transferrin receptor 2

Abstract

Transferrin receptor 2α (TfR2α), the major product of the TfRZ gene, is the second receptor for transferrin (Tf), which can mediate cellular iron uptake in vitro. Homozygous mutations of TfR2 cause haemochromatosis, suggesting that TfRZα may not be a simple iron transporter, but a regulator of iron by identifying iron-Tf. In this study, we analysed the ligand specificity of TfR2α using human transferrin receptor 1 (TfR1) and TfR2α-stably transfected and expressing cells and flow-cytometric techniques. We showed that human TfRZα interacted with both human and bovine Tf, whereas human TfR1 interacted only with human Tf. Neither human TfR1 nor TfRZα interacted with either lactoferrin or melanotransferrin. In addition, by creating point mutations in human TfRZα, the RGD sequence in the extracellular domain of TfRZα was shown to be crucial for Tf-binding. Furthermore, we demonstrated that mutated TfRZα (YZ50X), which has been reported in patients with hereditary haemochromatosis, also lost its ability to interact with both human and bovine Tf. Although human TfR1 and TfRZα share an essential structure (RGD) for ligand-binding, they have clearly different ligand specificities, which may be related to the differences in their roles in iron metabolism.