Production and characterization of functional recombinant hybrid heteropolymers of camel hepcidin and human ferritin H and L chains.
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.
['Boumaiza|Mohamed|M|', 'Carmona|Fernando|F|', 'Poli|Maura|M|', 'Asperti|Michela|M|', 'Gianoncelli|Alessandra|A|', 'Bertuzzi|Michela|M|', 'Ruzzenenti|Paola|P|', 'Arosio|Paolo|P|', 'Marzouki|Mohamed Nejib|MN|']
D000595:Amino Acid Sequence D000818:Animals D001052:Apoferritins D002162:Camelus D027682:Cation Transport Proteins D002460:Cell Line D064451:Hepcidins D006801:Humans D007501:Iron D051379:Mice D010084:Oxidation-Reduction D055503:Protein Multimerization D020836:Protein Structure, Quaternary D021381:Protein Transport D059748:Proteolysis D011993:Recombinant Fusion Proteins D012995:Solubility
heteropolymers camel hepcidin human ferritin H and L chains J774 cells