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Cited by 24Heat shock proteins researchNanopore and Nanochannel Transport StudiesForce Microscopy Techniques and Applications

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Single-molecule evidence of Entropic Pulling by Hsp70 chaperones

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Rukes, Verena, Rebeaud, Mathieu E., Perrin, Louis W., De Los Rios, Paolo, Cao, Chan·2024·Nature Communications
DOI 10.1038/s41467-024-52674-yPMC PMC11461734

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Abstract

Abstract Hsp70 chaperones are central components of the cellular network that ensure the structural quality of proteins. Despite their crucial roles in processes as diverse as the prevention of protein aggregation and protein translocation into organelles, their molecular mechanism of action has remained a hotly debated issue. Due to a lack of suitable methods, no experimental data has directly proven any of the models that have been proposed (Power Stroke, Brownian Ratchet, and Entropic Pulling). Recently, nanopores have emerged as a powerful tool to analyze the function of motor enzymes, as well as protein-protein interactions. Here, we used an in vitro single-molecule nanopore to mimic in vivo translocation of proteins, and to investigate the molecular mechanism of Hsp70. Our experiments demonstrate that Hsp70s forcefully extract polypeptide substrates that are trapped inside the pore. The forces they exert are strong at the molecular level, being equivalent to 46 pN over distances of 1 nm, and depend on the size of Hsp70. These findings provide unambiguous evidence supporting the Entropic Pulling mechanism of action of Hsp70s, thus solving a long-standing debate, and proposing a potentially universal principle governing diverse cellular processes. In addition, these results emphasize the utility of biological nanopores for studying protein function at the single-molecule level.

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Metadata

Authors
Rukes, V., Rebeaud, M. E., Perrin, L. W., De Los Rios, P., & Cao, C.
Affiliations
  • University of Geneva (CH)
  • École Polytechnique Fédérale de Lausanne (CH)
Corresponding author
Cao, Chan
ORCID
Venue
Nature Communications
Publisher
Springer Science and Business Media LLC
Published
2024-10-08
Volume
15
Issue
1
Pages
8604-8604
ISSN
2041-1723 (ISSN-L)
DOI
10.1038/s41467-024-52674-y
PMC
PMC11461734
OpenAlex
W4403228838
Cited by
24
References
1
Citations by year
2026: 7 · 2025: 16 · 2024: 1
FWCI
3.27
Type
journal article
Language
en
Open access
Licence
Locations
  • Nature Communications (publishedVersion, cc-by-nc-nd)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion)
  • PubMed Central (publishedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
  • Infoscience (Ecole Polytechnique Fédérale de Lausanne) (submittedVersion, cc-by-nc-nd)
  • RePEc: Research Papers in Economics (submittedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • Archive ouverte UNIGE (University of Geneva) (submittedVersion, other-oa)
Funders
Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung (PR00P3_193090)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Heat shock proteins research
  • Physical Sciences › Engineering › Biomedical Engineering › Nanopore and Nanochannel Transport Studies
  • Physical Sciences › Physics and Astronomy › Atomic and Molecular Physics, and Optics › Force Microscopy Techniques and Applications
Keywords
Hsp70 chaperones, protein disaggregation, protein translocation, power stroke, Brownian ratchet, single-molecule force measurement
MeSH terms
Single Molecule Imaging, Peptides, HSP70 Heat-Shock Proteins, Entropy, Escherichia coli Proteins, Nanopores

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