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Cotranslational folding of spectrin domains via partially structured states

Nature Structural & Molecular Biology volume 24, pages 221225 (2017) | Download Citation

Abstract

How do the key features of protein folding, elucidated from studies on native, isolated proteins, manifest in cotranslational folding on the ribosome? Using a well-characterized family of homologous α-helical proteins with a range of biophysical properties, we show that spectrin domains can fold vectorially on the ribosome and may do so via a pathway different from that of the isolated domain. We use cryo-EM to reveal a folded or partially folded structure, formed in the vestibule of the ribosome. Our results reveal that it is not possible to predict which domains will fold within the ribosome on the basis of the folding behavior of isolated domains; instead, we propose that a complex balance of the rate of folding, the rate of translation and the lifetime of folded or partly folded states will determine whether folding occurs cotranslationally on actively translating ribosomes.

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Acknowledgements

We thank A. Heuer for help preparing the figures. Supported by grants from the Swedish Cancer Foundation, the Swedish Research Council and the Knut and Alice Wallenberg Foundation (to G.v.H.); the Wellcome Trust (WT095195 to J.C.) and the European Research Council (ERC-2008-AdG 232648, to R.B.). J.C. is a Wellcome Trust Senior Research Fellow.

Author information

Author notes

    • Ola B Nilsson
    •  & Adrian A Nickson

    These authors contributed equally to this work.

Affiliations

  1. Department of Biochemistry and Biophysics Stockholm University, Stockholm, Sweden.

    • Ola B Nilsson
    •  & Gunnar von Heijne
  2. Department of Chemistry, University of Cambridge, Cambridge, UK.

    • Adrian A Nickson
    • , Jeffrey J Hollins
    • , Annette Steward
    •  & Jane Clarke
  3. Gene Center and Center for Integrated Protein Science Munich (CiPS-M), University of Munich, Munich, Germany.

    • Stephan Wickles
    •  & Roland Beckmann
  4. Science for Life Laboratory, Stockholm University, Solna, Sweden.

    • Gunnar von Heijne

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Contributions

O.B.N. and A.A.N. designed and carried out the experiments; J.J.H. and A.S. characterized the purified proteins; S.W. and R.B. were responsible for the cryo-EM experiments; A.S. wrote the manuscript; G.v.H. and J.C. conceived and planned the investigation and wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Gunnar von Heijne or Jane Clarke.

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    Supplementary Text and Figures

    Supplementary Figures 1–3 and Supplementary Table 1

Videos

  1. 1.

    Rigid body fit of the NMR structure of the R16 domain to the cryo-EM density map.

    Rigid body fit of the NMR structure of the R16 domain colored according to r.m.s. deviation (blue, 0.5–1.9; white, 2–3.9; red, ≥4.0 Å) to the cryo-EM density map (Fig. 4c).

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DOI

https://doi.org/10.1038/nsmb.3355