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The oligomeric structure of GroEL/GroES is required for biologically significant chaperonin function in protein folding

Nature Structural Biology volume 5, pages 977–985 (1998)Cite this article

An Erratum to this article was published on 01 February 1999

Abstract

Two models are being considered for the mechanism of chaperonin-assisted protein folding in E. coli: (i) GroEL/GroES act primarily by enclosing substrate polypeptide in a folding cage in which aggregation is prevented during folding. (ii) GroEL mediates the repetitive unfolding of misfolded polypeptides, returning them onto a productive folding track. Both models are not mutually exclusive, but studies with the polypeptide-binding domain of GroEL have suggested that unfolding is the primary mechanism, enclosure being unnecessary. Here we investigate the capacity of the isolated apical polypeptide-binding domain to functionally replace the complete GroEL/GroES system. We show that the apical domain binds aggregation-sensitive polypeptides but cannot significantly assist their refolding in vitro and fails to replace the groEL gene or to complement defects of groEL mutants in vivo. A single-ring version of GroEL cannot substitute for GroEL. These results strongly support the view that sequestration of aggregation-prone intermediates in a folding cage is an important element of the chaperonin mechanism.

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Figure 1: a, Ribbon diagram of the structure of a GroEL subunit (PDB file 1DER of NCBI)59 highlighting the apical, intermediate and equatorial domains.
Figure 2: Characterization of the apical domain, EL191–376, and of the domain fragment, N-His-EL193–334, by CD spectroscopy.
Figure 3: Interaction of apical domain proteins with unfolded rhodanese and DHFR.
Figure 4: Refolding of rhodanese, mMDH and CS in vitro in the presence of GroEL and apical domain proteins.
Figure 5: Effect of apical domain proteins on rhodanese reactivation by GroEL/GroES.
Figure 6: Rhodanese folding within the GroES-enclosed cavity of GroEL and GroEL-SR1.

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Acknowledgements

The authors thank L. Moroder and E. Weyher-Stingl (Martinsried, FRG) for their help with the CD measurements, and S. Radford (Leeds, UK) for stimulating discussion. We thank A. Fersht for suggesting the construction of the apical domain fragment lacking helices H11 and H12. This work was supported in part by Swiss National Fund. F.W. is a fellow of the German National Scholarship Foundation.

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Authors and Affiliations

  1. Max-Planck-Institut für Biochemie, Am Klopferspitz 18A, Martinsried, D-82152, Germany

    Frank Weber, Manajit K. Hayer-Hartl & F. Ulrich Hartl

  2. Departement de Biochimie Medicale, Centre Medical Universitaire , 1, rue Michel-Servet, Geneve , 1211, Switzerland

    France Keppel & Costa Georgopoulos

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Correspondence to F. Ulrich Hartl.

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Weber, F., Keppel, F., Georgopoulos, C. et al. The oligomeric structure of GroEL/GroES is required for biologically significant chaperonin function in protein folding. Nat Struct Mol Biol 5, 977–985 (1998). https://doi.org/10.1038/2952

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