EMDB/PDB/SASBDBから引用されている文献のデータベース

キーワード
構造解析手法	All X線回折 NMR 電子顕微鏡小角散乱中性子線回折線維回折粉末回折赤外分光 EPR FRET 理論モデルハイブリッド
著者・登録者
ジャーナル
IF	>30 >20 >10 >5 all

タイトル	Conformational changes during pore formation by the perforin-related protein pleurotolysin.
ジャーナル・号・ページ	PLoS Biol, Vol. 13, Issue 2, Page e1002049, Year 2015
掲載日	2015年2月5日
著者	Natalya Lukoyanova / Stephanie C Kondos / Irene Farabella / Ruby H P Law / Cyril F Reboul / Tom T Caradoc-Davies / Bradley A Spicer / Oded Kleifeld / Daouda A K Traore / Susan M Ekkel / Ilia Voskoboinik / Joseph A Trapani / Tamas Hatfaludi / Katherine Oliver / Eileen M Hotze / Rodney K Tweten / James C Whisstock / Maya Topf / Helen R Saibil / Michelle A Dunstone /
PubMed 要旨	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 ...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 Å 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 Å in diameter and 100 Å 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 ∼70° opening of the bent and distorted central β-sheet of the MACPF domain, accompanied by extrusion and refolding of two α-helical regions into transmembrane β-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 β-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 β-hairpins via top down zippering of backbone hydrogen bonds to form the membrane-inserted β-barrel. The intermediate structures of the MACPF domain during refolding into the β-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.
リンク	PLoS Biol / PubMed:25654333 / PubMed Central
手法	EM (単粒子) / X線回折
解像度	1.85 - 17.0 Å
構造データ	2793 4v2t EMDB-2793, PDB-4v2t: Membrane embedded pleurotolysin pore with 13 fold symmetry 手法: EM (単粒子) / 解像度: 11.0 Å 2794 4v3a EMDB-2794, PDB-4v3a: Membrane bound pleurotolysin prepore (TMH1 lock) trapped with engineered disulphide cross-link 手法: EM (単粒子) / 解像度: 15.0 Å 2795 4v3m EMDB-2795, PDB-4v3m: Membrane bound pleurotolysin prepore (TMH2 helix lock) trapped with engineered disulphide cross-link 手法: EM (単粒子) / 解像度: 17.0 Å 2796 4v3n EMDB-2796, PDB-4v3n: Membrane bound pleurotolysin prepore (TMH2 strand lock) trapped with engineered disulphide cross-link 手法: EM (単粒子) / 解像度: 14.0 Å PDB-4oeb: Structure of membrane binding protein pleurotolysin A from Pleurotus ostreatus 手法: X-RAY DIFFRACTION / 解像度: 1.85 Å PDB-4oej: Structure of membrane binding protein pleurotolysin B from Pleurotus ostreatus 手法: X-RAY DIFFRACTION / 解像度: 2.2 Å PDB-4ov8: Crystal Structure of the TMH1-lock mutant of the mature form of pleurotolysin B 手法: X-RAY DIFFRACTION / 解像度: 2.15 Å
化合物	ChemComp-SO4: SULFATE ION / 硫酸ジアニオン / 硫酸塩 ChemComp-HOH: WATER / 水 ChemComp-ACT: ACETATE ION / 酢酸イオン / 酢酸塩 ChemComp-GOL: GLYCEROL / グリセロ-ル / グリセリン ChemComp-CL: Unknown entry / クロリド / 塩化物
由来	pleurotus ostreatus (ヒラタケ)
キーワード	MEMBRANE BINDING PROTEIN / beta-sandwich fold / pleurotolysin B / MACPF domain / Pore formation / pleurotolysin A / TOXIN (毒素) / TMH1-lock / pore-forming protein (膜孔形成毒素) / Pleurtolysin A component / TRANSPORT PROTEIN (運搬体タンパク質) / MACPF/CDC SUPERFAMILY / PORE-FORMING PROTEINS (膜孔形成毒素)