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Dissection of the ATP-induced conformational cycle of the molecular chaperone Hsp90

Nature Structural & Molecular Biology volume 16, pages 287–293 (2009)Cite this article

Abstract

The molecular chaperone heat-shock protein 90 (Hsp90) couples ATP hydrolysis to conformational changes driving a reaction cycle that is required for substrate activation. Recent structural analysis provided snapshots of the open and closed states of Hsp90, which mark the starting and end points of these changes. Using fluorescence resonance energy transfer (FRET), we dissected the cycle kinetically and identified the intermediates on the pathway. The conformational transitions are orders of magnitude slower than the ATP-hydrolysis step and thus are the limiting events during the reaction cycle. Furthermore, these structural changes can be tightly regulated by cochaperones, being completely inhibited by Sti1 or accelerated by Aha1. In fact, even in the absence of nucleotide, Aha1 induces Hsp90 rearrangements that speed up the conformational cycle. This comprehensive reconstitution of the Hsp90 cycle defines a controlled progression through distinct intermediates that can be modulated by conformation-sensitive cochaperones.

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Figure 1: Establishment of the FRET system.
Figure 2: ATPγS-induced conformational changes.
Figure 3: Kinetic model of the ATPase cycle of Hsp90.
Figure 4: Conformational changes are coupled to N-terminal dimerization.
Figure 5: Cochaperone regulation of conformational changes.
Figure 6: Model of the Hsp90 cycle.

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Acknowledgements

This work was supported by grants from the Deutsche Forschungsgemeinschaft to J.B. (SFB594) and K.R. (RI1873-1/1) and the Fonds der Chemischen Industrie to J.B. and K.R. We thank E. Simpson, T. Hugel, M. Feige, T.M. Franzmann, J. Reinstein and C. Becker for carefully reading the manuscript.

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Authors and Affiliations

  1. Center for Integrated Protein Science at the Department Chemie, Technische Universität München, Lichtenbergstrasse 4, Garching, 85747, Germany

    Martin Hessling, Klaus Richter & Johannes Buchner

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  1. Martin Hessling
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  2. Klaus Richter
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  3. Johannes Buchner
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Contributions

M.H. performed experiments, data analysis and wrote the manuscript; K.R. performed data analysis and wrote the manuscript; J.B. designed experiments and wrote the manuscript.

Corresponding author

Correspondence to Johannes Buchner.

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Hessling, M., Richter, K. & Buchner, J. Dissection of the ATP-induced conformational cycle of the molecular chaperone Hsp90. Nat Struct Mol Biol 16, 287–293 (2009). https://doi.org/10.1038/nsmb.1565

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