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|Title:||Stepwise visualization of membrane pore formation by suilysin, a bacterial cholesterol-dependent cytolysin|
Dudkina, N. V.
Hodel, A. W.
Pandurangan, A. P.
Damaso, Mafalda Pires
Reboul, C. F.
Dunstone, M. A.
Andrew, Peter W.
Saibil, H. R.
Hoogenboom, B. W.
|Citation:||eLife , 2014;3:e04247|
|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.|
|Rights:||Copyright Leung et al. This article is distributed under the terms of the Creative Commons Attribution License http://creativecommons.org/licenses/by/4.0/, which permits unrestricted use and redistribution provided that the original author and source are credited.|
|Description:||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.|
|Appears in Collections:||Published Articles, Dept. of Infection, Immunity and Inflammation|
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|Stepwise visualization of membrane pore formation by suilysin, a bacterial cholesterol-dependent cytolysin.pdf||Published (publisher PDF)||2.71 MB||Adobe PDF||View/Open|
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