Production and characterization of functional recombinant hybrid heteropolymers of camel hepcidin and human ferritin H and L chains Boumaiza, Mohamed Carmona Rodríguez-Acosta, Fernando Poli, Maura Asperti, Michela Gianoncelli, Alessandra Bertuzzi, Michela Ruzzenenti, Paola Arosio, Paolo Marzouki, Mohamed Hepcidin Ferritin Heteropolymers Camel hepcidin Human ferritin H and L chains J774 cells This article has been accepted for publication in Protein Engineering design and Selection Published by Oxford University Press. 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. 2017-07-24T08:08:06Z 2017-07-24T08:08:06Z 2017-01 info:eu-repo/semantics/preprint Boumaiza, M.; et al. Production and characterization of functional recombinant hybrid heteropolymers of camel hepcidin and human ferritin H and L chains. Protein Engineering, Design and Selection 30(2): 77-84 (2017). [http://hdl.handle.net/10481/47248] 1741-0126 1741-0134 http://hdl.handle.net/10481/47248 10.1093/protein/gzw066 eng info:eu-repo/semantics/openAccess Oxford University Press