Letter | Published:

Alternative modes of client binding enable functional plasticity of Hsp70

Nature volume 539, pages 448451 (17 November 2016) | Download Citation


The Hsp70 system is a central hub of chaperone activity in all domains of life. Hsp70 performs a plethora of tasks, including folding assistance, protection against aggregation, protein trafficking, and enzyme activity regulation1,2,3,4,5, and interacts with non-folded chains, as well as near-native, misfolded, and aggregated proteins6,7,8,9,10. Hsp70 is thought to achieve its many physiological roles by binding peptide segments that extend from these different protein conformers within a groove that can be covered by an ATP-driven helical lid11,12,13,14,15. However, it has been difficult to test directly how Hsp70 interacts with protein substrates in different stages of folding and how it affects their structure. Moreover, recent indications of diverse lid conformations in Hsp70–substrate complexes raise the possibility of additional interaction mechanisms15,16,17,18. Addressing these issues is technically challenging, given the conformational dynamics of both chaperone and client, the transient nature of their interaction, and the involvement of co-chaperones and the ATP hydrolysis cycle19. Here, using optical tweezers, we show that the bacterial Hsp70 homologue (DnaK) binds and stabilizes not only extended peptide segments, but also partially folded and near-native protein structures. The Hsp70 lid and groove act synergistically when stabilizing folded structures: stabilization is abolished when the lid is truncated and less efficient when the groove is mutated. The diversity of binding modes has important consequences: Hsp70 can both stabilize and destabilize folded structures, in a nucleotide-regulated manner; like Hsp90 and GroEL, Hsp70 can affect the late stages of protein folding; and Hsp70 can suppress aggregation by protecting partially folded structures as well as unfolded protein chains. Overall, these findings in the DnaK system indicate an extension of the Hsp70 canonical model that potentially affects a wide range of physiological roles of the Hsp70 system.

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Work in the group of S.T. is supported by the Foundation for Fundamental Research on Matter (FOM) and the Netherlands Organization for Scientific Research (NWO). Work in the laboratory of B.B. was supported by research grants from the Deutsche Forschungsgemeinschaft (SFB638 and FOR1805) to G.K. and B.B. The work in the laboratory of M.P.M. was funded by the Deutsche Forschungsgemeinschaft (MA 1278/4-1). We thank M. M. Naqvi for performing control experiments, M. Avellaneda for help with preparing protein structure illustrations, and T. Shimizu, E. Garnett and F. Huber for critical reading of the manuscript.

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Author notes

    • Alireza Mashaghi
    •  & Sergey Bezrukavnikov

    These authors contributed equally to this work.


  1. FOM institute AMOLF, Science Park 104, 1098 XG Amsterdam, The Netherlands

    • Alireza Mashaghi
    • , Sergey Bezrukavnikov
    • , David P. Minde
    •  & Sander J. Tans
  2. Center for Molecular Biology of Heidelberg University (ZMBH), DKFZ-ZMBH Alliance, Im Neuenheimer Feld 282, D-69120 Heidelberg, Germany

    • Anne S. Wentink
    • , Roman Kityk
    • , Beate Zachmann-Brand
    • , Matthias P. Mayer
    • , Günter Kramer
    •  & Bernd Bukau
  3. German Cancer Research Center (DKFZ), Im Neuenheimer Feld 282, D-69120 Heidelberg, Germany

    • Anne S. Wentink
    • , Beate Zachmann-Brand
    • , Günter Kramer
    •  & Bernd Bukau


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A.M., S.B., D.M., M.P.M., G.K., B.B. and S.T. conceived and designed the research; B.Z.B. and D.M. designed and purified the MBP protein constructs; A.M. and S.B. performed the optical tweezers experiments; B.Z.B., G.K., A.W. and R.K. purified the DnaK system chaperones and DnaK mutants; A.W. performed the bulk RepE54 assays; R.K. performed the bulk luciferase assays; A.M., S.B., A.W. and S.T. analysed the data; and A.M., S.B., D.M., M.P.M., G.K., B.B. and S.T. wrote the paper.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Sander J. Tans.

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