4ov8: Difference between revisions
New page: '''Unreleased structure''' The entry 4ov8 is ON HOLD Authors: Kondos, S.C., Law, R.H.P., Whisstock, J.C., Dunstone, M.A. Description: Crystal Structure of the TMH1-lock mutant of the m... |
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The | ==Crystal Structure of the TMH1-lock mutant of the mature form of pleurotolysin B== | ||
<StructureSection load='4ov8' size='340' side='right'caption='[[4ov8]], [[Resolution|resolution]] 2.15Å' scene=''> | |||
== Structural highlights == | |||
<table><tr><td colspan='2'>[[4ov8]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Pleurotus_ostreatus Pleurotus ostreatus]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4OV8 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=4OV8 FirstGlance]. <br> | |||
</td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 2.15Å</td></tr> | |||
<tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=CL:CHLORIDE+ION'>CL</scene>, <scene name='pdbligand=GOL:GLYCEROL'>GOL</scene>, <scene name='pdbligand=SO4:SULFATE+ION'>SO4</scene></td></tr> | |||
<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=4ov8 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4ov8 OCA], [https://pdbe.org/4ov8 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=4ov8 RCSB], [https://www.ebi.ac.uk/pdbsum/4ov8 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=4ov8 ProSAT]</span></td></tr> | |||
</table> | |||
== Function == | |||
[https://www.uniprot.org/uniprot/Q5W9E8_PLEOS Q5W9E8_PLEOS] | |||
<div style="background-color:#fffaf0;"> | |||
== Publication Abstract from PubMed == | |||
Membrane attack complex/perforin-like (MACPF) proteins comprise the largest superfamily of pore-forming proteins, playing crucial roles in immunity and pathogenesis. Soluble monomers assemble into large transmembrane pores via conformational transitions that remain to be structurally and mechanistically characterised. Here we present an 11 A resolution cryo-electron microscopy (cryo-EM) structure of the two-part, fungal toxin Pleurotolysin (Ply), together with crystal structures of both components (the lipid binding PlyA protein and the pore-forming MACPF component PlyB). These data reveal a 13-fold pore 80 A in diameter and 100 A in height, with each subunit comprised of a PlyB molecule atop a membrane bound dimer of PlyA. The resolution of the EM map, together with biophysical and computational experiments, allowed confident assignment of subdomains in a MACPF pore assembly. The major conformational changes in PlyB are a approximately 70 degrees opening of the bent and distorted central beta-sheet of the MACPF domain, accompanied by extrusion and refolding of two alpha-helical regions into transmembrane beta-hairpins (TMH1 and TMH2). We determined the structures of three different disulphide bond-trapped prepore intermediates. Analysis of these data by molecular modelling and flexible fitting allows us to generate a potential trajectory of beta-sheet unbending. The results suggest that MACPF conformational change is triggered through disruption of the interface between a conserved helix-turn-helix motif and the top of TMH2. Following their release we propose that the transmembrane regions assemble into beta-hairpins via top down zippering of backbone hydrogen bonds to form the membrane-inserted beta-barrel. The intermediate structures of the MACPF domain during refolding into the beta-barrel pore establish a structural paradigm for the transition from soluble monomer to pore, which may be conserved across the whole superfamily. The TMH2 region is critical for the release of both TMH clusters, suggesting why this region is targeted by endogenous inhibitors of MACPF function. | |||
Conformational Changes during Pore Formation by the Perforin-Related Protein Pleurotolysin.,Lukoyanova N, Kondos SC, Farabella I, Law RH, Reboul CF, Caradoc-Davies TT, Spicer BA, Kleifeld O, Traore DA, Ekkel SM, Voskoboinik I, Trapani JA, Hatfaludi T, Oliver K, Hotze EM, Tweten RK, Whisstock JC, Topf M, Saibil HR, Dunstone MA PLoS Biol. 2015 Feb 5;13(2):e1002049. doi: 10.1371/journal.pbio.1002049., eCollection 2015 Feb. PMID:25654333<ref>PMID:25654333</ref> | |||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |||
</div> | |||
<div class="pdbe-citations 4ov8" style="background-color:#fffaf0;"></div> | |||
==See Also== | |||
*[[Pleurotolysin|Pleurotolysin]] | |||
== References == | |||
<references/> | |||
__TOC__ | |||
</StructureSection> | |||
[[Category: Large Structures]] | |||
[[Category: Pleurotus ostreatus]] | |||
[[Category: Dunstone MA]] | |||
[[Category: Kondos SC]] | |||
[[Category: Law RHP]] | |||
[[Category: Whisstock JC]] | |||
Latest revision as of 17:59, 8 November 2023
Crystal Structure of the TMH1-lock mutant of the mature form of pleurotolysin BCrystal Structure of the TMH1-lock mutant of the mature form of pleurotolysin B
Structural highlights
FunctionPublication Abstract from PubMedMembrane attack complex/perforin-like (MACPF) proteins comprise the largest superfamily of pore-forming proteins, playing crucial roles in immunity and pathogenesis. Soluble monomers assemble into large transmembrane pores via conformational transitions that remain to be structurally and mechanistically characterised. Here we present an 11 A resolution cryo-electron microscopy (cryo-EM) structure of the two-part, fungal toxin Pleurotolysin (Ply), together with crystal structures of both components (the lipid binding PlyA protein and the pore-forming MACPF component PlyB). These data reveal a 13-fold pore 80 A in diameter and 100 A in height, with each subunit comprised of a PlyB molecule atop a membrane bound dimer of PlyA. The resolution of the EM map, together with biophysical and computational experiments, allowed confident assignment of subdomains in a MACPF pore assembly. The major conformational changes in PlyB are a approximately 70 degrees opening of the bent and distorted central beta-sheet of the MACPF domain, accompanied by extrusion and refolding of two alpha-helical regions into transmembrane beta-hairpins (TMH1 and TMH2). We determined the structures of three different disulphide bond-trapped prepore intermediates. Analysis of these data by molecular modelling and flexible fitting allows us to generate a potential trajectory of beta-sheet unbending. The results suggest that MACPF conformational change is triggered through disruption of the interface between a conserved helix-turn-helix motif and the top of TMH2. Following their release we propose that the transmembrane regions assemble into beta-hairpins via top down zippering of backbone hydrogen bonds to form the membrane-inserted beta-barrel. The intermediate structures of the MACPF domain during refolding into the beta-barrel pore establish a structural paradigm for the transition from soluble monomer to pore, which may be conserved across the whole superfamily. The TMH2 region is critical for the release of both TMH clusters, suggesting why this region is targeted by endogenous inhibitors of MACPF function. Conformational Changes during Pore Formation by the Perforin-Related Protein Pleurotolysin.,Lukoyanova N, Kondos SC, Farabella I, Law RH, Reboul CF, Caradoc-Davies TT, Spicer BA, Kleifeld O, Traore DA, Ekkel SM, Voskoboinik I, Trapani JA, Hatfaludi T, Oliver K, Hotze EM, Tweten RK, Whisstock JC, Topf M, Saibil HR, Dunstone MA PLoS Biol. 2015 Feb 5;13(2):e1002049. doi: 10.1371/journal.pbio.1002049., eCollection 2015 Feb. PMID:25654333[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. See AlsoReferences
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