title: Production and characterization of functional recombinant hybrid heteropolymers of camel hepcidin and human ferritin H and L chains.
abstract: Hepcidin is a liver-synthesized hormone that plays a central role in the regulation of systemic iron homeostasis. To produce a new tool for its functional properties the cDNA coding for camel hepcidin-25 was cloned at the 5'end of human FTH sequence into the pASK-IBA43plus vector for expression in Escherichia coli The recombinant fusion hepcidin-ferritin-H subunit was isolated as an insoluble iron-containing protein. When alone it did not refold in a 24-mer ferritin molecule, but it did when renatured together with H- or L-ferritin chains. We obtained stable ferritin shells exposing about 4 hepcidin peptides per 24-mer shell. The molecules were then reduced and re-oxidized in a controlled manner to allow the formation of the proper hepcidin disulfide bridges. The functionality of the exposed hepcidin was confirmed by its ability to specifically bind the mouse macrophage cell line J774 that express ferroportin and to promote ferroportin degradation. This chimeric protein may be useful for studying the hepcidin-ferroportin interaction in cells and also as drug-delivery agent.
authors.affiliation: Laboratoire d'ingénierie des protéines et des molécules bioactives, Institut Nationale des Sciences Appliquées et de Technologie (I.N.S.A.T.) BP 676, Tunis Cedex 1080, Tunisie m.boumaiza@yahoo.fr.
authors.forename: Mohamed
authors.initials: M
authors.lastname: Boumaiza
authors.affiliation: Molecular Biology Laboratory, Department of Molecular and Translational Medicine, University of Brescia, Viale Europa 11 , Brescia, Italy.
authors.forename: Fernando
authors.initials: F
authors.lastname: Carmona
authors.affiliation: Molecular Biology Laboratory, Department of Molecular and Translational Medicine, University of Brescia, Viale Europa 11 , Brescia, Italy.
authors.forename: Maura
authors.initials: M
authors.lastname: Poli
authors.affiliation: Molecular Biology Laboratory, Department of Molecular and Translational Medicine, University of Brescia, Viale Europa 11 , Brescia, Italy.
authors.forename: Michela
authors.initials: M
authors.lastname: Asperti
authors.affiliation: Proteomics Platform, Department of Molecular and Translational Medicine, University of Brescia, Viale Europa 11, Brescia, Italy.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Gianoncelli
authors.affiliation: Proteomics Platform, Department of Molecular and Translational Medicine, University of Brescia, Viale Europa 11, Brescia, Italy.
authors.forename: Michela
authors.initials: M
authors.lastname: Bertuzzi
authors.affiliation: Molecular Biology Laboratory, Department of Molecular and Translational Medicine, University of Brescia, Viale Europa 11 , Brescia, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Ruzzenenti
authors.affiliation: Molecular Biology Laboratory, Department of Molecular and Translational Medicine, University of Brescia, Viale Europa 11 , Brescia, Italy.
authors.forename: Paolo
authors.initials: P
authors.lastname: Arosio
authors.affiliation: Laboratoire d'ingénierie des protéines et des molécules bioactives, Institut Nationale des Sciences Appliquées et de Technologie (I.N.S.A.T.) BP 676, Tunis Cedex 1080, Tunisie.
authors.forename: Mohamed Nejib
authors.initials: MN
authors.lastname: Marzouki
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001052:Apoferritins / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D002162:Camelus; D027682:Cation Transport Proteins / Q000378:metabolism; D002460:Cell Line; D064451:Hepcidins / Q000235:genetics*; D006801:Humans; D007501:Iron / Q000378:metabolism; D051379:Mice; D010084:Oxidation-Reduction; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D021381:Protein Transport; D059748:Proteolysis; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D012995:Solubility; D000098742:Ferroportin
chemical_list: D027682:Cation Transport Proteins; D064451:Hepcidins; D011993:Recombinant Fusion Proteins; D000098742:Ferroportin; D001052:Apoferritins; D007501:Iron
keywords: J774 cells; camel hepcidin; heteropolymers; human ferritin H and L chains