C-terminal amino acids are essential for human heat shock protein 70  dimerization.

Escherichia coli Proteins; HSP70 Heat-Shock Proteins; Recombinant Proteins; EC 3.6.1.- (dnaK protein, E coli); Circular Dichroism; Dimerization; Escherichia coli/metabolism; Escherichia coli Proteins/chemistry/metabolism; HSP70 Heat-Shock Proteins/chemistry/genetics/metabolism; Humans; Isoelectric Point; Protein Refolding; Protein Structure, Secondary; Recombinant Proteins/biosynthesis/chemistry/isolation & purification; Spectrometry, Fluorescence

Abstract :

[en] 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.

Disciplines :

Biochemistry, biophysics & molecular biology

Author, co-author :

Marcion, Guillaume ; Université de Liège - ULiège > GIGA > GIGA I3 - Hematology ; INSERM, UMR 866, 7 blvd Jeanne d'Arc, 21000, Dijon, France.

Seigneuric, Renaud

Chavanne, Evelyne

Artur, Yves

Briand, Loïc

Hadi, Tarik

Gobbo, Jessica

Garrido, Carmen

Neiers, Fabrice

Language :

English

Title :

C-terminal amino acids are essential for human heat shock protein 70 dimerization.

Publication date :

January 2015

Journal title :

Cell Stress and Chaperones

ISSN :

1355-8145

eISSN :

1466-1268

Publisher :

Cell Stress Society International, Us ct

Volume :

Issue :

Pages :

61-72

Peer reviewed :

Peer Reviewed verified by ORBi

Available on ORBi :

since 25 May 2022

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