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Retrieving high-resolution information from disordered 2D crystals by single-particle cryo-EM

Author

Listed:
  • Ricardo D. Righetto

    (Biozentrum, University of Basel)

  • Nikhil Biyani

    (Biozentrum, University of Basel)

  • Julia Kowal

    (Biozentrum, University of Basel
    Institute for Molecular Biology and Biophysics, ETH Zürich)

  • Mohamed Chami

    (Biozentrum, University of Basel)

  • Henning Stahlberg

    (Biozentrum, University of Basel)

Abstract

Electron crystallography can reveal the structure of membrane proteins within 2D crystals under close-to-native conditions. High-resolution structural information can only be reached if crystals are perfectly flat and highly ordered. In practice, such crystals are difficult to obtain. Available image unbending algorithms correct for disorder, but only perform well on images of non-tilted, flat crystals, while out-of-plane distortions are not addressed. Here, we present an approach that employs single-particle refinement procedures to locally unbend crystals in 3D. With this method, density maps of the MloK1 potassium channel with a resolution of 4 Å were obtained from images of 2D crystals that do not diffract beyond 10 Å. Furthermore, 3D classification allowed multiple structures to be resolved, revealing a series of MloK1 conformations within a single 2D crystal. This conformational heterogeneity explains the poor diffraction observed and is related to channel function. The approach is implemented in the FOCUS package.

Suggested Citation

  • Ricardo D. Righetto & Nikhil Biyani & Julia Kowal & Mohamed Chami & Henning Stahlberg, 2019. "Retrieving high-resolution information from disordered 2D crystals by single-particle cryo-EM," Nature Communications, Nature, vol. 10(1), pages 1-10, December.
    • Handle: RePEc:nat:natcom:v:10:y:2019:i:1:d:10.1038_s41467-019-09661-5
    DOI: 10.1038/s41467-019-09661-5
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    Cited by:

    1. Eliza S. Nieweglowska & Axel F. Brilot & Melissa Méndez-Moran & Claire Kokontis & Minkyung Baek & Junrui Li & Yifan Cheng & David Baker & Joseph Bondy-Denomy & David A. Agard, 2023. "The ϕPA3 phage nucleus is enclosed by a self-assembling 2D crystalline lattice," Nature Communications, Nature, vol. 14(1), pages 1-12, December.

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