Transferrin receptor complex

Many of the investigations into endosomal pathways have concentrated on receptor-mediated endocytosis, as in the iron-transferrin-receptor complex, and it is not clear how the systems vary depending on whether or not the pathway is clathrin-dependent or clathrin-independent [54],... 

Iron ion The protein transferrin binds ferric ions and transports them in the semm aronnd the body. The ions are taken np by cells via a transferrin receptor which is present in the plasma membrane. The receptor binds transferrin and the complex enters the cell where transferrin releases... 

The uptake of iron into the cell could follow several pathways. The iron could be released from the transferrin at the receptor site and be carried into the cell. Alternatively, the whole transferrin-receptor complex could be taken into the cell via endocytosis, and passed into an acidic compartment, where the iron is released, passed out of the compartment, and stored in ferritin. 

It should be pointed out, however, that not all hormones dissociate from their receptor in the pH 5.5 environment of the endosome [24], Some hormone-receptor complexes require much lower pH values for dissociation to occur. Although not a peptide hormone, the iron-transport protein transferrin is a peculiar example of this phenomenon and should be pointed out. In this case, at the neutral pH of the extracellular fluid transferrin containing bound iron binds to its cell surface receptor and is internalized. In the low pH environment of the endosome, iron becomes dissociated from transferrin, but transferrin remains bound to its receptor. The transferrin receptor, with bound transferrin, is then recycled to the cell surface. With iron no longer bound to the transferrin, the transferrin readily dissociates from its receptor at the neutral pH of the extracellular fluid [25,26]. This mechanism provides for an efficient continual uptake of iron into cells. Unlike transferrin, however, in those instances where peptide hormones have been documented not to be dissociated from their receptor in the endosome compartment, the hormone and receptor are delivered to the lysosomes via fusion of the endosomes with lyso-somes, where both hormone and receptor are degraded [24,27]. The continuous degradation of the receptor with each round of RME eventually leads to a decrease in the number of receptors on the cell surface, a phenomenon called down-regulation. 

As noted above, iron-loaded serum transferrin has the role of transporting Fe(III) to cells requiring it. Once it reaches a target cell, it binds to the transferrin receptor (TfR) on the cell outer surface. TfR is a disulfide-linked dimer that binds two transferrin molecules. At neutral pH, apo transferrin does not bind to the transferrin receptor. Once formed, the transferrin receptor complex becomes detached from the cell membrane and enters the cell enclosed in a clathrin-coated vesicle. Uncoating of the vesicle generates an endosome in which the pH is lowered to 5.5, promoting release of iron. At this pH, the transferrin remains bound to its receptor, and... 

Optical spectra of transferrin C-lobe docked with the transferrin receptor showed a characteristic broad absorption band centred at 465 nm, just as in the receptor-free /zo/o-protein (Figure 2.1 inset). The intensity of this absorbance band declined as more negative potentials were applied in a spectroelectrochemistry experiment, but did not qualitatively change in its overall features. An EPR spectrum of the Fec/TfR complex at pH 5.8, recovered from the OTTLE cell after completion of spectroelectrochemical studies allowed us to conclude that the first coordination shell of Fe " in transferrin is intact and unperturbed when C-lobe is complexed with TfR. Consequently, we assume that C-lobe and Fec/TfR complex have similar if not identical Fe " and Fe binding constants, and so we take for binding of Fe " in the protein-receptor complex to be 10 M as calculated for free Tf. This value was used to correct the observed Nernst plot data by accounting for the dissociation of Fe that occurs upon reduction. Nernst plots for the observed spectroelectrochemical data for FccTf/TfR, and data corrected for Fe dissociation, are presented in Figure 2.7. The corrected plot exhibits typical Nernstian behaviour for a one-electron transfer and a E1/2 value of —285 mV. 

Figure 2.8 compares corrected Nernst plots for C-lobe half-transferrin free in solution and bound to the transferrin receptor, at endosomal pH. These data clearly demonstrate that docking iron-loaded C-lobe transferrin at the transferrin receptor at pH 5.8 makes it energetically more favourable to reduce Fe " to Fe by 200 mV. Furthermore, receptor-docking places Fe reduction in a range accessible to NADH or NADPH cofactors, consistent with the hypothesis that reduction is the initial event in iron release from transferrin in the endosome. Fe " is bound by /zTf at least 14 orders of magnitude more weakly than Fe, so that reductive release of iron bound to HTi in the transferrin-transferrin receptor complex is then physiologically and thermodynamically feasible, and the barrier to transport across the endosomal membrane is lifted. The transferrin receptor, therefore, is more than a simple conveyor of... 

Several strategies for increasing the permeability of the brain capillaries to proteins have been developed. The permeability of the BBB can be transiently increased by intra-arterial injection of the solutions with high osmolarity, which disrupts inter-endothelial tight junctions [11]. Certain protein modifications, such as cationization by hexamethyldiamine [12] and anionization by succinylation [13], produce enhanced uptake in the brain. Modification of drugs [14] and proteins [15] by linkage to an anti-transferrin receptor antibody also appears to enhance transport into the brain. This approach depends on receptor-mediated transcytosis of transferrin-receptor complexes by brain endothelial cells substantial uptake also occurs in the liver. 

In humans, iron is transported across the gut by a series of poorly defined processes. Fe(III), ferric ion, is absorbed via a J03 integrin and mobilferrin, whereas ferrous ion enter the cells via Nramp. Once inside the body, Fe(III) is transported through the serum by transferrin, a protein of molecular weight 63,000 Da. Fe(III) transferrin is recognized by a receptor protein on the cell surface. Via a process known as cell-mediated endocytosis, the Fe(III) transferrin/receptor complex induces the external cell membrane to pucker and eventually form a clatharin-coated vesicle in the cytoplasm. After removal of the clatharin, the vesicle (known as an endosome) becomes... 

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