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-Structure paper
タイトル | High-resolution structure of the Shigella type-III secretion needle by solid-state NMR and cryo-electron microscopy. |
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ジャーナル・号・ページ | Nat Commun, Vol. 5, Page 4976, Year 2014 |
掲載日 | 2014年9月29日 |
著者 | Jean-Philippe Demers / Birgit Habenstein / Antoine Loquet / Suresh Kumar Vasa / Karin Giller / Stefan Becker / David Baker / Adam Lange / Nikolaos G Sgourakis / |
PubMed 要旨 | We introduce a general hybrid approach for determining the structures of supramolecular assemblies. Cryo-electron microscopy (cryo-EM) data define the overall envelope of the assembly and rigid-body ...We introduce a general hybrid approach for determining the structures of supramolecular assemblies. Cryo-electron microscopy (cryo-EM) data define the overall envelope of the assembly and rigid-body orientation of the subunits while solid-state nuclear magnetic resonance (ssNMR) chemical shifts and distance constraints define the local secondary structure, protein fold and inter-subunit interactions. Finally, Rosetta structure calculations provide a general framework to integrate the different sources of structural information. Combining a 7.7-Å cryo-EM density map and 996 ssNMR distance constraints, the structure of the type-III secretion system needle of Shigella flexneri is determined to a precision of 0.4 Å. The calculated structures are cross-validated using an independent data set of 691 ssNMR constraints and scanning transmission electron microscopy measurements. The hybrid model resolves the conformation of the non-conserved N terminus, which occupies a protrusion in the cryo-EM density, and reveals conserved pore residues forming a continuous pattern of electrostatic interactions, thereby suggesting a mechanism for effector protein translocation. |
リンク | Nat Commun / PubMed:25264107 / PubMed Central |
手法 | NMR (固体) / EM (らせん対称) |
解像度 | 7.7 Å |
構造データ | PDB-2mme: |
由来 |
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キーワード | PROTEIN TRANSPORT / type-three secretion system / filamentous protein / helical assembly / Shigella flexneri / protein translocation / hybrid methods / Rosetta |