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Chaperonin complex with a newly folded protein encapsulated in the folding chamber

Nature volume 457, pages 107–110 (2009)Cite this article

Abstract

A subset of essential cellular proteins requires the assistance of chaperonins (in Escherichia coli, GroEL and GroES), double-ring complexes in which the two rings act alternately to bind, encapsulate and fold a wide range of nascent or stress-denatured proteins1,2,3,4,5. This process starts by the trapping of a substrate protein on hydrophobic surfaces in the central cavity of a GroEL ring6,7,8,9,10. Then, binding of ATP and co-chaperonin GroES to that ring ejects the non-native protein from its binding sites, through forced unfolding or other major conformational changes, and encloses it in a hydrophilic chamber for folding11,12,13,14,15. ATP hydrolysis and subsequent ATP binding to the opposite ring trigger dissociation of the chamber and release of the substrate protein3. The bacteriophage T4 requires its own version of GroES, gp31, which forms a taller folding chamber, to fold the major viral capsid protein gp23 (refs 16–20). Polypeptides are known to fold inside the chaperonin complex, but the conformation of an encapsulated protein has not previously been visualized. Here we present structures of gp23–chaperonin complexes, showing both the initial captured state and the final, close-to-native state with gp23 encapsulated in the folding chamber. Although the chamber is expanded, it is still barely large enough to contain the elongated gp23 monomer, explaining why the GroEL–GroES complex is not able to fold gp23 and showing how the chaperonin structure distorts to enclose a large, physiological substrate protein.

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Figure 1: Asymmetric reconstructions of GroEL with non-native gp23 in one or both rings.
Figure 2: Asymmetric reconstructions of GroEL–gp31 without visible substrate, with gp23 in the open ring and with gp23 in both rings.
Figure 3: Folding chambers of the GroEL–gp23–gp31 complexes.
Figure 4: Substrate densities isolated from the binary and ternary complexes.

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Accession codes

Data deposits

The three-dimensional reconstructions for the two binary complexes and the three ternary complexes are deposited in the EBI-MSD EMD database with accession codes EMD-1544 through to EMD-1548.

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Acknowledgements

We thank R. Westlake, D. Houldershaw and L. Wang for computing and EM support. This work was carried out at the School of Crystallography, Birkbeck College, and was supported by a Wellcome Trust programme grant, EU 3D EM Network of Excellence and 3D Repertoire grants.

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Author notes

  1. P. J. Bakkes

    Present address: Present address: Department of Molecular Microbiology, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Kerklaan 30, 9751 NN Haren, The Netherlands.,

Authors and Affiliations

  1. Department of Crystallography and Institute for Structural and Molecular Biology, Birkbeck College, Malet Street, London WC1E 7HX, UK,

    D. K. Clare & H. R. Saibil

  2. Department of Pathology, VU University Medical Center, De Boelelaan 1117, 1081 HV Amsterdam, The Netherlands,

    P. J. Bakkes, H. van Heerikhuizen & S. M. van der Vies

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  1. D. K. Clare
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Corresponding authors

Correspondence to S. M. van der Vies or H. R. Saibil.

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This file contains Supplementary Figures 1-3 with Legends, Supplementary Table 1, Supplementary Notes and Discussion and Supplementary References (PDF 1537 kb)

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Clare, D., Bakkes, P., van Heerikhuizen, H. et al. Chaperonin complex with a newly folded protein encapsulated in the folding chamber. Nature 457, 107–110 (2009). https://doi.org/10.1038/nature07479

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