Binding Chromium(III) to Form Mixed Cr(III),Fe(III) Serum Transferrins

Transferrin, Tf, the protein that transports iron as Fe(III) from the blood to the tissues via endocytosis, is believed to also transport Cr(III). Under physiological conditions, Tf binds and releases Cr(III) rapidly from Cr(III)2-Tf; however, the major form of Tf in the bloodstream is mono-ferric Tf (Fe(III)-Tf). Given the low concentration of Cr(III) in the bloodstream, the form of Cr(III)-containing Tf that is transported is probably monochromic, monoferric-Tf (Cr(III),Fe(III)-Tf). Given that Tf has two specific metal-binding sites, one in both its C-terminal and its N-terminal lobe, two forms of Cr(III),Fe(III)-Tf can form. The binding of Cr(III) to mono-ferric Tf to generate both forms of Cr(III),Fe(III)-Tf has been examined in detail for the first time. The addition of Cr(III) to monoferric Tfs in 100 mM HEPES and 25 mM bicarbonate solution, pH 7.4, resulted in a rapid binding of Cr(III) to the open metal-binding site of the Tfs. Titrations of the monoferric Tfs with Cr(III) indicate the tight binding on one Cr(III) in each case. The binding of Cr(III) to monoferric Tfs is accompanied by conformational changes similar to adding two equivalents of Cr(III) to apoTf. Thus, mono-ferric Tfs bind one equivalent of Cr(III) rapidly and tightly to form mixed Cr(III),Fe(III)-Tfs. Cr(III) is probably transported as mixed Cr(III),Fe(III)-Tfs.