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The client protein p53 adopts a molten globule–like state in the presence of Hsp90

Nature Structural & Molecular Biology volume 18, pages 537–541 (2011)Cite this article

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Abstract

It is not currently known in what state (folded, unfolded or alternatively folded) client proteins interact with the chaperone Hsp90. We show that one client, the p53 DNA-binding domain, undergoes a structural change in the presence of Hsp90 to adopt a molten globule–like state. Addition of one- and two-domain constructs of Hsp90, as well as the full-length three-domain protein, to isotopically labeled p53 led to reduction in NMR signal intensity throughout p53, particularly in its central β-sheet. This reduction seems to be associated with a change of structure of p53 without formation of a distinct complex with Hsp90. Fluorescence and hydrogen-exchange measurements support a loosening of the structure of p53 in the presence of Hsp90 and its domains. We propose that Hsp90 interacts with p53 by multiple transient interactions, forming a dynamic heterogeneous manifold of conformational states that resembles a molten globule.

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Figure 1: Schematic of domains of Hsp90 and p53.
Figure 2: 1H-15N spectra of 15N-labeled p53 with added Hsp90.
Figure 3: p53 resonance attenuation by Hsp90 proteins.
Figure 4: Schematic of model for NMR titration results.
Figure 5: ANS fluorescence spectra and H/D exchange.

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Acknowledgements

We thank M. Kostic for preparation of p53 and Hsp90 expression constructs and preliminary binding assays. We thank P. Wright and members of the Wright and Dyson groups for helpful comments, G. Kroon for help with NMR experiments and E. Manlapaz for technical assistance. The original clone used to prepare the domains of human Hsp90α was provided by D. Toft of the Mayo Clinic (Rochester, Minnesota, USA). This work was supported by grant GM57374 from the US National Institutes of Health, and by a grant from the Korea Research Foundation (KRF-2006-214-E0009), funded by the Korean Government (MOEHRD).

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  1. Sung Jean Park

    Present address: Present address: College of Pharmacy, Gachon University School of Medicine and Science, Incheon, Korea.,

Authors and Affiliations

  1. Department of Molecular Biology, The Scripps Research Institute, La Jolla, California, USA

    Sung Jean Park, Brendan N Borin, Maria A Martinez-Yamout & H Jane Dyson

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  1. Sung Jean Park
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Contributions

S.J.P. and H.J.D. designed experiments; S.J.P. carried out NMR and fluorescence experiments; B.N.B. carried out H/D exchange experiments; S.J.P., M.A.M.-Y. and H.J.D. analyzed data; S.J.P., M.A.M.-Y. and H.J.D. wrote the paper.

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Correspondence to H Jane Dyson.

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Supplementary Figures 1–5, Supplementary Table 1 and Supplementary Discussion (PDF 592 kb)

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Park, S., Borin, B., Martinez-Yamout, M. et al. The client protein p53 adopts a molten globule–like state in the presence of Hsp90. Nat Struct Mol Biol 18, 537–541 (2011). https://doi.org/10.1038/nsmb.2045

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