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Chaperones convert the energy from ATP into the nonequilibrium stabilization of native proteins

Nature Chemical Biology volume 14, pages 388–395 (2018)Cite this article

Abstract

During and after protein translation, molecular chaperones require ATP hydrolysis to favor the native folding of their substrates and, under stress, to avoid aggregation and revert misfolding. Why do some chaperones need ATP, and what are the consequences of the energy contributed by the ATPase cycle? Here, we used biochemical assays and physical modeling to show that the bacterial chaperones GroEL (Hsp60) and DnaK (Hsp70) both use part of the energy from ATP hydrolysis to restore the native state of their substrates, even under denaturing conditions in which the native state is thermodynamically unstable. Consistently with thermodynamics, upon exhaustion of ATP, the metastable native chaperone products spontaneously revert to their equilibrium non-native states. In the presence of ATPase chaperones, some proteins may thus behave as open ATP-driven, nonequilibrium systems whose fate is only partially determined by equilibrium thermodynamics.

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Fig. 1: Free-energy landscape of MDH at 37 °C and schematic action of chaperones.
Fig. 2: ATP-dependent nonequilibrium GroELS recovery and maintenance of MDH activity in denaturing conditions.
Fig. 3: Results from the model.
Fig. 4: ATP-driven catalytic action of substoichiometric amounts of GroELS on MDH at 37 °C.
Fig. 5: GroELS converts the energy of ATP into a nonequilibrium stabilization of the native state of MDH.

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Acknowledgements

We thank summer-project students P. Inns and I. Koog for setting up the citrate synthase enzymatic assay and performing key preliminary and control chaperone assays. This project was financed in part by Swiss National Science Foundation grants 140512/1 and 31003A_156948, and Swiss State Secretariat for Education Research and Innovation grant C15.0042 to P.G. and B.F., and by Swiss National Science Foundation grant 200020_163042 to P.D.L.R. and A.S.S. A.B. acknowledges support from the French Agence Nationale de la Recherche (ANR), under grant ANR-14-ACHN-0016. We thank H.-J. Schönfeld (F. Hoffmann-La Roche) for providing GrpE.

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Authors and Affiliations

  1. Department of Plant Molecular Biology, Université de Lausanne, Lausanne, Switzerland

    Pierre Goloubinoff & Bruno Fauvet

  2. Institute of Physics, School of Basic Sciences, École Polytechnique Fédérale de Lausanne (EPFL), Lausanne, Switzerland

    Alberto S. Sassi & Paolo De Los Rios

  3. Centre de Biochimie Structurale (CBS), INSERM, CNRS, Université de Montpellier, Montpellier, France

    Alessandro Barducci

  4. Institute of Bioengineering, School of Life Sciences, EPFL, Lausanne, Switzerland

    Paolo De Los Rios

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  1. Pierre Goloubinoff
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  5. Paolo De Los Rios
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Contributions

P.G., A.B. and P.D.L.R. conceived the study. P.G. and B.F. performed the experiments. P.D.L.R., A.S.S. and A.B. developed the model. P.D.L.R. and A.S.S. performed the calculations. All authors contributed to the writing of the work.

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Correspondence to Pierre Goloubinoff or Paolo De Los Rios.

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Goloubinoff, P., Sassi, A.S., Fauvet, B. et al. Chaperones convert the energy from ATP into the nonequilibrium stabilization of native proteins. Nat Chem Biol 14, 388–395 (2018). https://doi.org/10.1038/s41589-018-0013-8

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