University of Leicester
Stepwise visualization of membrane pore formation by suilysin, a bacterial cholesterol-dependent cytolysin.pdf (2.65 MB)

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

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journal contribution
posted on 2015-03-11, 09:52 authored by C. Leung, N. V. Dudkina, N. Lukoyanova, A. W. Hodel, I. Farabella, A. P. Pandurangan, Nasrin Jahan, Mafalda Pires Damaso, D. Osmanovic, C. F. Reboul, M. A. Dunstone, Peter W. Andrew, Rana Lonnen, M. Topf, H. R. Saibil, B. W. Hoogenboom
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.


BBSRC (BB/G011729/1, BB/J005932/1, BB/J006254 and BB/ K01692X/1), the ERC (advanced grant 294408), and the Leverhulme Trust (RPG-2012-519).



eLife , 2014;3:e04247

Author affiliation

/Organisation/COLLEGE OF MEDICINE, BIOLOGICAL SCIENCES AND PSYCHOLOGY/School of Medicine/Department of Infection, Immunity and Inflammation


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Figure supplement 1. Symmetry of suilysin prepores and pores, determined by negative-stain EM. DOI: 10.7554/eLife.04247.004 Figure supplement 2. Resolution curves for EM maps. DOI: 10.7554/eLife.04247.005 Figure supplement 3. Electrostatic potential maps and interacting residues. DOI: 10.7554/eLife.04247.006 Figure supplement 4. Comparison between prepore and crystal structure conformations. DOI: 10.7554/eLife.04247.007 Accession codes for EM maps: prepore EMD-12698, pore EMD-12711.