Transferrins sites

Box 12-F), which tends to occupy a fraction of the transferrin sites in blood, although it binds much more weakly than does Fe. Several other binding sites for transition metals are pictured in Chapter 16. 

Excess iron citrate reagent is added to the sample to saturate all available transferrin sites. After an incubation period of three to five minutes the treated specimen is applied to an alumina column where iron that is not bound to transferrin is absorbed. The transferrin-bound iron contained in the eluate is the total iron-binding capacity of the specimen. The total iron-binding capacity is determined with the slides for iron in the same manner as for an untreated serum specimen. (For details of the iron method, see pp. 197 fl). 

Transferrin (Tf) is a plasma protein that plays a centtal role in transporting iron around the body to sites where... 

Transferrin Shuttles Iron to Sites Where It Is Needed... 

Transferrin (Tf) is a Pj-globulin with a molecular mass of approximately 76 kDa. it is a glycoprotein and is synthesized in the liver. About 20 polymorphic forms of transferrin have been found, it plays a central role in the body s metabolism of iron because it transports iron (2 mol of Fe + per mole of Tf) in the circulation to sites where iron is required, eg, from the gut to the bone marrow and other organs. Approximately 200 billion red blood cells (about 20 mL) are catabolized per day, releasing about 25 mg of iron into the body—most of which will be transported by transferrin. 

HFE has been shown to be located in cells in the crypts of the small intestine, the site of iron absorption. There is evidence that it associates with P2 niicroglobu-lin, an association that may be necessary for its stability, intracellular processing, and cell surface expression. The complex interacts with the transferrin receptor (TfR) how this leads to excessive storage of iron when HFE is altered by mutation is under close smdy. The mouse homolog of HFE has been knocked out, resulting in a potentially useful animal model of hemochromatosis. 

Transferrin binds iron, transporting it to sites where it is required. Ferritin provides an intracellular store of iron. Iron deficiency anemia is a very prevalent disorder. Hereditary hemochromatosis has been shown to be due to mutations in HFE, a gene encoding the protein HFE, which appeats to play an important role in absorption of iron. 

Lactoferrin resembles transferrin in terms of molecular weight, amino-acid sequence homology and number of Fe(lII) binding sites. Lactoferrin is released from activated PMNs upon degranulation and may play a role in myelopoiesis, primary antibody response, lymphocyte proliferation, cytokine production and complement activation. 

Figure 5.5 Stereo view of the Fe3+ binding site of (a) hFBP (b) human lactoferrin, N-lobe and (c) human transferrin (N-lobe). From Bruns, 1997. Reproduced by permission of Nature Publishing Group. 

Figure 5.7 Structures of the alternative conformations of the iron-binding sites in the orthorhombic crystal form of the recombinant N-lobe of human transferrin. Reproduced with permission from MacGillivray et ah, 1998. Copyright (1998), American Chemical Society. 

The determination of the structure of the iron transporter, ferric-binding, protein (hFBP)t from Haemophilus influenzae (Bruns et ah, 1997) at 0.16 nm resolution shows that it is a member of the transferrin superfamily, which includes both the transferrins and a number of periplasmic binding proteins (PBP). The PBPs transport a wide variety of nutrients, including sugars, amino acids and ions, across the periplasm from the outer to the inner (plasma) membrane in bacteria (see Chapter 3). Iron binding by transferrins (see below) requires concomitant binding of a carbonate anion, which is located at the N-terminus of a helix. This corresponds to the site at which the anions are specifically bound in the bacterial periplasmic sulfate- and... 

The iron-binding sites have been characterized by crystallographic studies on several transferrins, and in Figure 5.7 (Plate 7) that of the N-lobe of human lactoferrin is presented. The 3+ charge on the ferric ion is matched by the three anionic ligands Asp-63, Tyr-95 and Tyr-188 (the fourth, His-249, is neutral), while the charge on the carbonate anion is almost matched by the positive charge on Arg-124 and the... 

A recently obtained high resolution structure of two crystal forms of the N-lobe of human serum transferrin (at 0.16 and 0.18 nm resolution) shows disorder at the iron-binding sites (MacGillivray et ah, 1998). Model building and refinement show... 

The equivalent of the tryptic fragment of human transferrin receptor has been expressed in Chinese hamster ovary cells and its structure determined at a resolution of 0.32 nm (Lawrence et ah, 1999). The asymmetric unit of the crystals contains four transferrin receptor dimers. Interpretable electron density is found for the entire tryptic fragment except for Arg-121 at the amino terminus, and density is also seen for the first N-acetylglucosamine residue at each of the N-glycosylation sites. The transferrin receptor monomer is made up of three distinct domains, organized such that the dimer is butterfly shaped (Figure 5.10, Plate 7). The likely orientation of the dimer with respect to the plasma membrane has been assigned on the basis of the... 

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