TRANSFERRIN, A MECHANISM FOR IRON RELEASE

Iron release from transferrin has been investigated in mildly acidic and acidic media in the presence of formate, acetate and citrate. It occurs first from the N-terminal iron-binding site (N-site) of the holoprotein. It is independent of the nature and the concentration of competing ligands and is controlled by a slow proton transfer; second-order rate constant k(1) = (7.4 +/- 0.5) X 10(4) M(-1) s(-1) which can be attributed to a rate-limiting slow proton gain by a protein Ligand subsequent to a fast decarbonation of the N-site. Iron loss from the C-terminal iron-binding site (C-site) is slower than that from the N-site and occurs by two pathways. The first is favoured below pH 4 and does not involve the formation of an intermediate ternary complex. It can be controlled by a rate-limiting slow proton-triggered decarbonation of the binding site; second-order rate constant k(3) = (2.25 +/- 0.05) X 10(4) M(-1) s(-1). The second pathway is favoured above pH 4 and involves a mixed protein-ligand iron complex. It takes place through the slow protonation of the mixed ternary complex and depends on the nature of the competing ligand. It is faster in the presence of citrate than in that of acetate; second-order rate constant k(4) = (1.75 +/- 0.10) X 10(3) M(-1) s(-1) for citrate and (85 +/- 5) M(-1) s(-1) for acetate. All these phenomena can possibly describe proton-triggered changes of conformation of the binding sites.