Expression and purification of a new recombinant camel hepcidin able to promote the degradation of the iron exporter ferroportin1

Autor: Jean-Christophe Deschemin, Mohamed Nejib Marzouki, Aymen Ezzine, Noureddine Ben Khalaf, Sophie Vaulont, Maryse Jaouen, Marie Agnès Sari, Mohamed Boumaiza
Přispěvatelé: Institut National des Sciences Appliquées et de Technologie [Tunis] (INSAT), Laboratoire de Chimie et de Biochimie Pharmacologiques et Toxicologiques (LCBPT - UMR 8601), Université Paris Descartes - Paris 5 (UPD5)-Centre National de la Recherche Scientifique (CNRS), Institut Cochin (IC UM3 (UMR 8104 / U1016)), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Laboratoire d'Immunopathologie, Vaccinologie et Génétique Moléculaire (LVGM), Institut Pasteur de Tunis, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP), This work is entirely financed by the Tunisian Ministry of High Education, Scientific Research and Technology and the Laboratoryof Protein Engineering and Bioactive Molecules (LIP-MB) in theNational Institute of Applied Sciences and Technology of Tunis,University of Carthage. Special thanks, for their contribution in this work, to the research group of the Proteomics Platform of theUniversity Paris Descartes (3P5), Université Paris Descartes - Paris 5 (UPD5)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Université Paris Descartes - Paris 5 (UPD5) - Centre National de la Recherche Scientifique (CNRS), Institut Cochin (UM3 (UMR 8104 / U1016)), Université Paris Descartes - Paris 5 (UPD5) - Institut National de la Santé et de la Recherche Médicale (INSERM) - Centre National de la Recherche Scientifique (CNRS), Institut Pasteur de Tunis - Réseau International des Instituts Pasteur (RIIP)
Jazyk: angličtina
Rok vydání: 2015
Předmět:
[SDV]Life Sciences [q-bio]
Ferroportin
Hepcidin
Peptide
medicine.disease_cause
law.invention
Mice
0302 clinical medicine
law
hemic and lymphatic diseases
Disulfides
Internalization
Cation Transport Proteins
media_common
Leishmania major
chemistry.chemical_classification
0303 health sciences
biology
Recombinant Proteins
[SDV] Life Sciences [q-bio]
Biochemistry
030220 oncology & carcinogenesis
Recombinant DNA
[SDV.MP.PAR] Life Sciences [q-bio]/Microbiology and Parasitology/Parasitology
Biotechnology
Plasmids
inorganic chemicals
endocrine system
congenital
hereditary
and neonatal diseases and abnormalities
Camelus
media_common.quotation_subject
Molecular Sequence Data
03 medical and health sciences
Hepcidins
medicine
Escherichia coli
Animals
Humans
[SDV.MP.PAR]Life Sciences [q-bio]/Microbiology and Parasitology/Parasitology
Amino Acid Sequence
Disulfide bridge
030304 developmental biology
Macrophages
nutritional and metabolic diseases
Fusion protein
[SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology
chemistry
Spectrometry
Mass
Matrix-Assisted Laser Desorption-Ionization
biology.protein
[SDV.MP.BAC] Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology
Cysteine
Zdroj: Protein Expression and Purification
Protein Expression and Purification, Elsevier, 2015, 115, pp.11-18. ⟨10.1016/j.pep.2015.04.016⟩
Protein Expression and Purification, 2015, 115, pp.11-18. ⟨10.1016/j.pep.2015.04.016⟩
Protein Expression and Purification, Elsevier, 2015, 115, pp.11-18. 〈10.1016/j.pep.2015.04.016〉
ISSN: 1046-5928
1096-0279
DOI: 10.1016/j.pep.2015.04.016⟩
Popis: International audience; 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 ferroportinl (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. (C) 2015 Elsevier Inc. All rights reserved.
Databáze: OpenAIRE
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