Stepwise visualization of membrane pore formation by suilysin, a bacterial cholesterol-dependent cytolysin

Carl Leung, Natalya V Dudkina, Natalya Lukoyanova, Adrian W. Hodel, Irene Farabella, Arun P. Pandurangan, Nasrin Jahan, Mafalda Pires Damaso, Dino Osmanović, Cyril F Reboul, Michelle A Dunstone, Peter W. Andrew, Rana Lonnen, Maya Topf, Helen Ruth Saibil, Bart W. Hoogenboom

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76 Citations (Scopus)

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.

Original languageEnglish
Article numbere04247
Number of pages17
JournaleLife
Volume3
DOIs
Publication statusPublished - 2014

Keywords

  • bacterial toxins
  • biophysics
  • cholesterol-dependent cytolysins
  • membrane pore formation
  • pore-forming proteins
  • S. suis
  • structural biology

Cite this

Leung, C., Dudkina, N. V., Lukoyanova, N., Hodel, A. W., Farabella, I., Pandurangan, A. P., Jahan, N., Pires Damaso, M., Osmanović, D., Reboul, C. F., Dunstone, M. A., Andrew, P. W., Lonnen, R., Topf, M., Saibil, H. R., & Hoogenboom, B. W. (2014). Stepwise visualization of membrane pore formation by suilysin, a bacterial cholesterol-dependent cytolysin. eLife, 3, [e04247]. https://doi.org/10.7554/eLife.04247