A fluorescent multi-domain protein reveals the unfolding mechanism of Hsp70.
Détails
Télécharger: 36266349_BIB_C8354A2A8BC2.pdf (5685.58 [Ko])
Etat: Public
Version: Final published version
Licence: CC BY 4.0
Etat: Public
Version: Final published version
Licence: CC BY 4.0
ID Serval
serval:BIB_C8354A2A8BC2
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
A fluorescent multi-domain protein reveals the unfolding mechanism of Hsp70.
Périodique
Nature chemical biology
ISSN
1552-4469 (Electronic)
ISSN-L
1552-4450
Statut éditorial
Publié
Date de publication
02/2023
Peer-reviewed
Oui
Volume
19
Numéro
2
Pages
198-205
Langue
anglais
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: ppublish
Publication Status: ppublish
Résumé
Detailed understanding of the mechanism by which Hsp70 chaperones protect cells against protein aggregation is hampered by the lack of a comprehensive characterization of the aggregates, which are typically heterogeneous. Here we designed a reporter chaperone substrate, MLucV, composed of a stress-labile luciferase flanked by stress-resistant fluorescent domains, which upon denaturation formed a discrete population of small aggregates. Combining Förster resonance energy transfer and enzymatic activity measurements provided unprecedented details on the aggregated, unfolded, Hsp70-bound and native MLucV conformations. The Hsp70 mechanism first involved ATP-fueled disaggregation and unfolding of the stable pre-aggregated substrate, which stretched MLucV beyond simply unfolded conformations, followed by native refolding. The ATP-fueled unfolding and refolding action of Hsp70 on MLucV aggregates could accumulate native MLucV species under elevated denaturing temperatures highly adverse to the native state. These results unambiguously exclude binding and preventing of aggregation from the non-equilibrium mechanism by which Hsp70 converts stable aggregates into metastable native proteins.
Mots-clé
Protein Folding, HSP70 Heat-Shock Proteins/chemistry, Molecular Chaperones/chemistry, Luciferases/metabolism, Adenosine Triphosphate, Protein Denaturation, Protein Unfolding
Pubmed
Web of science
Open Access
Oui
Création de la notice
02/11/2022 9:11
Dernière modification de la notice
23/01/2024 7:34