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Pulling geometry defines the mechanical resistance of a β-sheet protein

Nature Structural & Molecular Biology volume 10, pages 731–737 (2003)Cite this article

An Erratum to this article was published on 01 October 2003

Abstract

Proteins show diverse responses when placed under mechanical stress. The molecular origins of their differing mechanical resistance are still unclear, although the orientation of secondary structural elements relative to the applied force vector is thought to have an important function. Here, by using a method of protein immobilization that allows force to be applied to the same all-β protein, E2lip3, in two different directions, we show that the energy landscape for mechanical unfolding is markedly anisotropic. These results, in combination with molecular dynamics (MD) simulations, reveal that the unfolding pathway depends on the pulling geometry and is associated with unfolding forces that differ by an order of magnitude. Thus, the mechanical resistance of a protein is not dictated solely by amino acid sequence, topology or unfolding rate constant, but depends critically on the direction of the applied extension.

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Figure 1: Structure of E2lip3 and its inclusion in concatamers of I27.
Figure 2: Force-extension curves of different concatamers.
Figure 3: Mechanical unfolding statistics of (I27)5, (I27)4E2lip3(+) and (I27)4E2lip3(−) at a tip-retraction speed of 700 nm s−1.
Figure 4: Pulling-speed dependence of unfolding forces for (I27)4E2lip3(+) and (I27)5* (a heteropolymer of I27 domains studied previously7).
Figure 5: Force-extension profiles for (I27)4E2lip3(+) can be used to obtain kinetic information.
Figure 6: Force-extension profiles in a series of SMD simulations and structures at different time points.

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Acknowledgements

We thank A. Berry for help with producing Figure 1 and K. Ainley for the large-scale microbial cultures. We thank A. Blake and the other members of the Radford laboratory for fruitful discussions. We acknowledge the Biotechnology and Biological Sciences Research Council, Engineering and Physical Sciences Research Council, University of Leeds, the Wellcome Trust and Forschungskredit der Universität Zürich for financial support. S.E.R. is a BBSRC Professorial Fellow. The manuscript is a contribution from the Astbury Centre for Structural Molecular Biology, which is part of the North of England Structural Biology Centre (NESBIC) and is funded by the BBSRC.

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Authors and Affiliations

  1. School of Biochemistry and Molecular Biology, University of Leeds, LS2 9JT, UK

    David J Brockwell, Rebecca C Zinober & Sheena E Radford

  2. Biochemisches Institut der Univerität Zürich, Winterthurerstrasse 190, Zürich, CH-8057, Switzerland

    Emanuele Paci

  3. Department of Physics and Astronomy, University of Leeds, Leeds, LS2 9JT, UK

    Rebecca C Zinober, Peter D Olmsted & D Alastair Smith

  4. School of Chemistry, University of Leeds, Leeds, LS2 9JT, UK

    Godfrey S Beddard

  5. Department of Biochemistry, University of Cambridge, Cambridge, CB2 1GA, UK

    Richard N Perham

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Correspondence to Sheena E Radford.

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Brockwell, D., Paci, E., Zinober, R. et al. Pulling geometry defines the mechanical resistance of a β-sheet protein. Nat Struct Mol Biol 10, 731–737 (2003). https://doi.org/10.1038/nsb968

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