Fiche publication
Date publication
janvier 2015
Journal
Cell stress & chaperones
Auteurs
Membres identifiés du Cancéropôle Est :
Dr GARRIDO Carmen
,
Dr GOBBO Jessica
Tous les auteurs :
Marcion G, Seigneuric R, Chavanne E, Artur Y, Briand L, Hadi T, Gobbo J, Garrido C, Neiers F
Lien Pubmed
Résumé
The human inducible heat shock protein 70 (hHsp70), which is involved in several major pathologies, including neurodegenerative disorders and cancer, is a key molecular chaperone and contributes to the proper protein folding and maintenance of a large number of protein structures. Despite its role in disease, the current structural knowledge of hHsp70 is almost exclusively based on its Escherichia coli homolog, DnaK, even though these two proteins only share ~50 % amino acid identity. For the first time, we describe a complete heterologous production and purification strategy that allowed us to obtain a large amount of soluble, full-length, and non-tagged hHsp70. The protein displayed both an ATPase and a refolding activity when combined to the human Hsp40. Multi-angle light scattering and bio-layer interferometry analyses demonstrated the ability of hHsp70 to homodimerize. The role of the C-terminal part of hHsp70 was identified and confirmed by a study of a truncated version of hHsp70 that could neither dimerize nor present refolding activity.
Mots clés
Circular Dichroism, Dimerization, Escherichia coli, metabolism, Escherichia coli Proteins, chemistry, HSP70 Heat-Shock Proteins, chemistry, Humans, Isoelectric Point, Protein Refolding, Protein Structure, Secondary, Recombinant Proteins, biosynthesis, Spectrometry, Fluorescence
Référence
Cell Stress Chaperones. 2015 Jan;20(1):61-72
