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Journal Article

High-resolution structure of the Shigella type-III secretion needle by solid-state NMR and cryo-electron microscopy.

MPS-Authors
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Demers,  J. P.
Research Group of Solid-State NMR, MPI for biophysical chemistry, Max Planck Society;

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Habenstein,  B.
Research Group of Solid-State NMR, MPI for biophysical chemistry, Max Planck Society;

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Loquet,  A.
Research Group of Solid-State NMR, MPI for biophysical chemistry, Max Planck Society;

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Vasa,  S. K.
Research Group of Solid-State NMR, MPI for biophysical chemistry, Max Planck Society;

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Giller,  K.
Department of NMR-Based Structural Biology, MPI for biophysical chemistry, Max Planck Society;

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Becker,  S.
Department of NMR-Based Structural Biology, MPI for biophysical chemistry, Max Planck Society;

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Lange,  A.
Research Group of Solid-State NMR, MPI for biophysical chemistry, Max Planck Society;

Fulltext (public)

2070006.pdf
(Publisher version), 3MB

Supplementary Material (public)

2070006_Suppl.pdf
(Supplementary material), 691KB

Citation

Demers, J. P., Habenstein, B., Loquet, A., Vasa, S. K., Giller, K., Becker, S., et al. (2014). High-resolution structure of the Shigella type-III secretion needle by solid-state NMR and cryo-electron microscopy. Nature Communications, 5: 4976. doi:10.1038/ncomms5976.


Cite as: http://hdl.handle.net/11858/00-001M-0000-0024-291B-3
Abstract
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-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 angstrom. 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.