title: Evidence of H- and L-chains have co-operative roles in the iron-uptake mechanism of human ferritin.
abstract: The ability to incorporate iron in vitro was studied in homopolymers of human ferritin L-chain, human ferritin H-chain and its variants and in homopolymer mixtures. The H-chain variants carried amino acid substitutions in the ferroxidase centre and/or in carboxy residues on the cavity surface. Iron incorporation was examined by gel electrophoresis of the reaction products by staining for iron and protein. It was found that inactivation of the ferroxidase centre combined with the substitution of four carboxy groups on the cavity abolished the ability of H-chain ferritin to incorporate iron. Competition experiments with limited amounts of iron showed that, at neutral pH, L-chain ferritin is more efficient in forming iron cores than the H-chain variants altered at the ferroxidase activity or in the cavity. Competition experiments at pH 5.5 demonstrated that L-chain apoferritin is able to incorporate iron only when in the presence of H-chain variants with ferroxidase activity. The results indicate that L-chain apoferritin has a higher capacity than the H-chain apoferritin to induce iron-core nucleation, whereas H-chain ferritin is superior in promoting Fe(II) oxidation. The finding of cooperative roles of the H- and L-chains in ferritin iron uptake provides a clue to understanding the biological function of isoferritins.
authors.affiliation: Department of Biomedical Science and Technology, San Raffaele Institute, University of Milano, Italy.
authors.forename: S
authors.initials: S
authors.lastname: Levi
authors.forename: S J
authors.initials: SJ
authors.lastname: Yewdall
authors.forename: P M
authors.initials: PM
authors.lastname: Harrison
authors.forename: P
authors.initials: P
authors.lastname: Santambrogio
authors.forename: A
authors.initials: A
authors.lastname: Cozzi
authors.forename: E
authors.initials: E
authors.lastname: Rovida
authors.forename: A
authors.initials: A
authors.lastname: Albertini
authors.forename: P
authors.initials: P
authors.lastname: Arosio
mesh_terms: D001052:Apoferritins / Q000378:metabolism; D001667:Binding, Competitive; D005293:Ferritins / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D007501:Iron / Q000378:metabolism*; D046911:Macromolecular Substances; D008958:Models, Molecular; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D013329:Structure-Activity Relationship
chemical_list: D046911:Macromolecular Substances; D011994:Recombinant Proteins; D005293:Ferritins; D001052:Apoferritins; D007501:Iron