1. Biophysics and Structural Biology
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Stepwise visualization of membrane pore formation by suilysin, a bacterial cholesterol-dependent cytolysin

  1. Carl Leung
  2. Natalya V Dudkina
  3. Natalya Lukoyanova
  4. Adrian W Hodel
  5. Irene Farabella
  6. Arun P Pandurangan
  7. Nasrin Jahan
  8. Mafalda Pires Damaso
  9. Dino Osmanović
  10. Cyril F Reboul
  11. Michelle A Dunstone
  12. Peter W Andrew
  13. Rana Lonnen
  14. Maya Topf
  15. Helen R Saibil
  16. Bart W Hoogenboom Is a corresponding author
  1. University College London, United Kingdom
  2. Birkbeck College, United Kingdom
  3. University of Leicester, United Kingdom
  4. Monash University, Australia
Research Article
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Cite as: eLife 2014;3:e04247 doi: 10.7554/eLife.04247

Abstract

Membrane attack complex/perforin/cholesterol-dependent cytolysin (MACPF/CDC) proteins constitute a major superfamily of pore-forming proteins that act as bacterial virulence factors and effectors in immune defence. Upon binding to the membrane, they convert from the soluble monomeric form to oligomeric, membrane-inserted pores. Using real-time atomic force microscopy (AFM), electron microscopy (EM) and atomic structure fitting, we have mapped the structure and assembly pathways of a bacterial CDC in unprecedented detail and accuracy, focussing on suilysin from Streptococcus suis. We show that suilysin assembly is a noncooperative process that is terminated before the protein inserts into the membrane. The resulting ring-shaped pores and kinetically trapped arc-shaped assemblies are all seen to perforate the membrane, as also visible by the ejection of its lipids. Membrane insertion requires a concerted conformational change of the monomeric subunits, with a marked expansion in pore diameter due to large changes in subunit structure and packing.

Article and author information

Author details

  1. Carl Leung

    1. London Centre for Nanotechnology, University College London, London, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  2. Natalya V Dudkina

    1. Department of Crystallography, Birkbeck College, London, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  3. Natalya Lukoyanova

    1. Department of Crystallography, Birkbeck College, London, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  4. Adrian W Hodel

    1. London Centre for Nanotechnology, University College London, London, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  5. Irene Farabella

    1. Department of Crystallography, Birkbeck College, London, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  6. Arun P Pandurangan

    1. Department of Crystallography, Birkbeck College, London, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  7. Nasrin Jahan

    1. Department of Infection, Immunity, and Inflammation, University of Leicester, Leicester, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  8. Mafalda Pires Damaso

    1. Department of Infection, Immunity, and Inflammation, University of Leicester, Leicester, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  9. Dino Osmanović

    1. London Centre for Nanotechnology, University College London, London, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  10. Cyril F Reboul

    1. Department of Biochemistry and Molecular Biology, Monash University, Melbourne, Australia
    Competing interests
    The authors declare that no competing interests exist.
  11. Michelle A Dunstone

    1. Department of Biochemistry and Molecular Biology, Monash University, Melbourne, Australia
    Competing interests
    The authors declare that no competing interests exist.
  12. Peter W Andrew

    1. Department of Infection, Immunity, and Inflammation, University of Leicester, Leicester, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  13. Rana Lonnen

    1. Department of Infection, Immunity, and Inflammation, University of Leicester, Leicester, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  14. Maya Topf

    1. Department of Crystallography, Birkbeck College, London, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  15. Helen R Saibil

    1. Department of Crystallography, Birkbeck College, London, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  16. Bart W Hoogenboom

    1. London Centre for Nanotechnology, University College London, London, United Kingdom
    For correspondence
    1. b.hoogenboom@ucl.ac.uk
    Competing interests
    The authors declare that no competing interests exist.

Reviewing Editor

  1. Volker Dötsch, Reviewing Editor, Goethe University, Germany

Publication history

  1. Received: August 5, 2014
  2. Accepted: November 24, 2014
  3. Accepted Manuscript published: December 2, 2014 (version 1)
  4. Version of Record published: December 24, 2014 (version 2)

Copyright

© 2014, Leung et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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