title: Expression and purification of a new recombinant camel hepcidin able to promote the degradation of the iron exporter ferroportin1.
abstract: Hepcidin, a 25-amino-acid and highly disulfide bonded antimicrobial peptide, is the central regulator of iron homeostasis. This hormone is expressed in response to iron and inflammation and interacts with ferroportin1 (FPN1), the only known iron exporter in vertebrates, inducing its internalization and degradation. Thus, the export of iron from cells to plasma will be significantly diminished. Thereby, hepcidin has become the target of intense research studies due to its profound biomedical significance. This study describes the functional expression of recombinant camel hepcidin in Escherichia coli. Biologically active recombinant camel hepcidin was obtained thanks to the production of a hepcidin-thioredoxin fusion protein (TRX-HepcD) and a purified camel hepcidin, with an extra methionine at the N-terminus, was obtained after enterokinase cleavage of the fusion protein. Presence of the four disulfide bridges was verified using MALDI-ToF spectrometry. The recombinant camel hepcidin was compared to related synthetic bioactive peptides, including human hepcidin, and was found equally able to promote ferroportin degradation of mouse macrophages. Furthermore, camel hepcidins exhibits a high capacity to inhibit the growth of Leishmania major promastigotes. These results proved that production of functional camel hepcidin can be achieved in E. coli, this is a major interest for the production of cysteine rich peptides or proteins that can be purified under their functional form without the need of a refolding process.
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
authors.initials: M
authors.lastname: Boumaiza
authors.affiliation: Laboratoire de Chimie et Biochimie Pharmacologiques et Toxicologiques, CNRS UMR 8601, Université Paris Descartes, Sorbonne Paris Cité, 45 rue des Saints Pères, 75006 Paris, France.
authors.forename: Maryse
authors.initials: M
authors.lastname: Jaouen
authors.affiliation: Institut Cochin, Inserm U1016, CNRS (UMR 8104), Université Paris Descartes, Sorbonne Paris Cité, Paris, France.
authors.forename: Jean-Christophe
authors.initials: JC
authors.lastname: Deschemin
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: Aymen
authors.initials: A
authors.lastname: Ezzine
authors.affiliation: Laboratoire d'Immunopathologie, vaccinologie et génétique moléculaire (LIVGM), Institut Pasteur de Tunis, 13 Place Pasteur, 1002 Tunis-Belvédère, Tunisie.
authors.forename: Noureddine
authors.initials: N
authors.lastname: Ben Khalaf
authors.affiliation: Institut Cochin, Inserm U1016, CNRS (UMR 8104), Université Paris Descartes, Sorbonne Paris Cité, Paris, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Vaulont
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 Nèjib
authors.initials: MN
authors.lastname: Marzouki
authors.affiliation: Laboratoire de Chimie et Biochimie Pharmacologiques et Toxicologiques, CNRS UMR 8601, Université Paris Descartes, Sorbonne Paris Cité, 45 rue des Saints Pères, 75006 Paris, France. Electronic address: marie-agnes.sari@parisdescartes.fr.
authors.forename: Marie Agnès
authors.initials: MA
authors.lastname: Sari
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002162:Camelus / Q000235:genetics; D027682:Cation Transport Proteins / Q000737:chemistry / Q000378:metabolism*; D004220:Disulfides / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics; D064451:Hepcidins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D006801:Humans; D008264:Macrophages / Q000737:chemistry / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D010957:Plasmids; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D000098742:Ferroportin
chemical_list: D027682:Cation Transport Proteins; D004220:Disulfides; D064451:Hepcidins; D011994:Recombinant Proteins; D000098742:Ferroportin
keywords: Disulfide bridge; Ferroportin; Hepcidin; Leishmania major; Peptide