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Identification of in vivo substrates of the chaperonin GroEL

Nature volume 402, pages 147–154 (1999)Cite this article

Abstract

The chaperonin GroEL has an essential role in mediating protein folding in the cytosol of Escherichia coli. Here we show that GroEL interacts strongly with a well-defined set of approximately 300 newly translated polypeptides, including essential components of the transcription/translation machinery and metabolic enzymes. About one third of these proteins are structurally unstable and repeatedly return to GroEL for conformational maintenance. GroEL substrates consist preferentially of two or more domains with αβ-folds, which contain α-helices and buried β-sheets with extensive hydrophobic surfaces. These proteins are expected to fold slowly and be prone to aggregation. The hydrophobic binding regions of GroEL may be well adapted to interact with the non-native states of αβ-domain proteins.

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Figure 1: 2D-gel analysis of newly translated proteins that transit through GroEL.
Figure 2: Kinetics and extent of release from GroEL of newly translated proteins.
Figure 3: Pre-existing proteins that cycle on GroEL.
Figure 4: Identification of GroEL substrates by large-scale immunoprecipitation.
Figure 5: Structural classification of GroEL substrates.

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Acknowledgements

We thank K. Andersson for technical assistance and M. C. Bewley for help with Fig. 5b.

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Authors and Affiliations

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

    Walid A. Houry & F. Ulrich Hartl

  2. GSF-Forschungszentrum für Umwelt und Gesundheit, Munich Information Center for Protein Sequences, Max-Planck-Institut für Biochemie, Am Klopferspitz 18A, Martinsried, D-82152, Germany

    Dmitrij Frishman

  3. Department of Protein Analytics, Max-Planck-Institut für Biochemie, Am Klopferspitz 18A, Martinsried, D-82152, Germany

    Friedrich Lottspeich

  4. Proteomics Division, Toplab GmbH, Fraunhoferstrasse 18A, Martinsried, D-82152, Germany

    Christoph Eckerskorn

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Correspondence to F. Ulrich Hartl.

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Houry, W., Frishman, D., Eckerskorn, C. et al. Identification of in vivo substrates of the chaperonin GroEL. Nature 402, 147–154 (1999). https://doi.org/10.1038/45977

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