Production and characterization of functional recombinant hybrid heteropolymers of camel hepcidin and human ferritin H and L chains
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AutorBoumaiza, Mohamed; Carmona Rodríguez-Acosta, Fernando; Poli, Maura; Asperti, Michela; Gianoncelli, Alessandra; Bertuzzi, Michela; Ruzzenenti, Paola; Arosio, Paolo; Marzouki, Mohamed
Oxford University Press
HepcidinFerritinHeteropolymersCamel hepcidinHuman ferritin H and L chainsJ774 cells
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]
PatrocinadorThis work is partially financed by the Laboratory of Protein Engineering and Bioactive Molecules (LIP-MB) and the Doctoral School of the National Institute of Applied Sciences and Technology (INSAT-Tunis) – University of Carthage.
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.