Ferric ion sequestering agents: kinetics of iron release from ferritin to catechoylamides

The removal of ferric ion from the iron storage protein ferritin to synthetic catechoylamide sequestering agents has been studied using visible spectroscopy at 487 nm. One ligand which has been investigated in detail is N,N',N' ',-tris(2,3-dihydroxy-5-sulfobenzoyl)-1,5,10-triazadecane (3,4-LICAMS), which octahedrally coordinates the metal ion via six phenolic oxygens. For some related catechoylamide chelates, the percentage of iron removed after 6 h has been determined. These ligands incorporate various modifications, either on the catechol moiety or on the backbone structure of the ligand. Mobilization of iron by the catechoylamide ligands alone results in very slow exchange, and virtually no iron removal after 6 h. In contrast, addition of ascorbic acid to the reaction mixture facilitates iron exchange, with the release of 7% of the available iron in the same time span. Variation of the initial rate with ascorbic acid concentration results in Michaelis-Menten kinetics with Km = 1.7 . 10(-3) M and a maximal rate of 1.28 . 10(-7) M . min-1. The ascorbic acid-mediated rate was not affected by changing the catechoylamide ligand concentration, and was only slightly affected by variation of the ligand employed. These data are consistent with a multistep process which includes diffusion of a reductant into the ferritin inner core, reduction and possible chelation of the ferrous ion, diffusion out of the protein shell, and subsequent iron exchange with the catechoylamide molecule.