title: Iron uptake in ferritin is blocked by binding of [Cr(TREN)(H(2)O)(OH)](2+), a slow dissociating model for [Fe(H(2)O)(6)](2+).
abstract: Ferritin concentrates iron as a hydrous ferric oxide in a protein cavity (8 nm in diameter) by using eight pores along the threefold symmetry axes of the octahedral supramolecular structure. The role of ligand exchange in the entry of Fe(II) hexahydrate into ferritin protein has been studied with [Cr(TREN)(H(2)O)(OH)](2+) [TREN = N(CH(2)CH(2)NH(2))(3)], a model for Fe(H(2)O)(6)2+ with only two exchangeable ligands. The results show that five different ferritin proteins, varying in pore structure, oxidation sites, and nucleation sites, bind Cr(TREN) at functional protein sites, based on inhibition of iron mineralization and oxidation. Properties of Cr(TREN)-ferritin adducts include an increased isoelectric point, a shift in the Cr(TREN) UV/vis spectrum consistent with exchange of water for protein carboxylate or thiolate ligands, binding affinities of 50-250 microM, and a slow rate of dissociation (k = 4 x 10(-6) sec(-1)). The relationship of Cr(TREN) inhibition of iron oxidation and mineralization by Cr(TREN) to the known structures of the various ferritins tested suggests that Cr(TREN) plugs the ferritin pores, obstructing Fe(II) entry in folded and unfolded pores. Because only two exchangeable waters are sufficient for pore binding of Cr(TREN), the physiological Fe(II) donor must bind to the pore with few exchangeable ligands. These results show the advantage of using stable model complexes to explore properties of transient Fe-protein complexes during Fe mineralization in ferritin.
authors.affiliation: Department of Chemistry, University of California, Berkeley, CA 94720-1460, USA.
authors.forename: Carmen M
authors.initials: CM
authors.lastname: Barnés
authors.forename: Elizabeth C
authors.initials: EC
authors.lastname: Theil
authors.forename: Kenneth N
authors.initials: KN
authors.lastname: Raymond
mesh_terms: D001665:Binding Sites; D004305:Dose-Response Relationship, Drug; D005293:Ferritins / Q000737:chemistry*; D007501:Iron / Q000737:chemistry / Q000493:pharmacokinetics*; D007700:Kinetics; D008024:Ligands; D008956:Models, Chemical; D008958:Models, Molecular; D009942:Organometallic Compounds / Q000737:chemistry*; D010100:Oxygen / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D013053:Spectrophotometry; D013997:Time Factors; D014466:Ultraviolet Rays
chemical_list: D008024:Ligands; D009942:Organometallic Compounds; C456278:aquahydroxo(tris(aminoethyl)amine)chromium(III); D005293:Ferritins; D007501:Iron; D010100:Oxygen