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  Structural characterization of supramolecular assemblies by 13C spin dilution and 3D solid-state NMR.

Habenstein, B., Loquet, A., Giller, K., Becker, S., & Lange, A. (2013). Structural characterization of supramolecular assemblies by 13C spin dilution and 3D solid-state NMR. Journal of biomolecular NMR, 55(1), 1-9. doi:10.1007/s10858-012-9691-9.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-000E-E517-0 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0028-DD4A-6
Genre: Journal Article

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 Creators:
Habenstein, B.1, Author              
Loquet, A.1, Author              
Giller, K.2, Author              
Becker, S.2, Author              
Lange, A.1, Author              
Affiliations:
1Research Group of Solid-State NMR, MPI for biophysical chemistry, Max Planck Society, ou_persistent35              
2Department of NMR Based Structural Biology, MPI for biophysical chemistry, Max Planck Society, ou_578567              

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Free keywords: Solid-state NMR; Isotopic labeling; Resonance assignment; Structure determination; Secretion systems; Supramolecular assemblies
 Abstract: 13C spin diluted protein samples can be produced using [1-13C] and [2-13C]-glucose (Glc) carbon sources in the bacterial growth medium. The 13C spin dilution results in favorable 13C spectral resolution and polarization transfer behavior. We recently reported the combined use of [1-13C]- and [2-13C]-Glc labeling to facilitate the structural analysis of insoluble and non-crystalline biological systems by solid-state NMR (ssNMR), including sequential assignment, detection of long-range contacts and structure determination of macromolecular assemblies. In solution NMR the beneficial properties of sparsely labeled samples using [2-13C]-glycerol (13C labeled Cα sites on a 12C diluted background) have recently been exploited to provide a bi-directional assignment method (Takeuchi et al. in J Biomol NMR 49(1):17–26, 2011 ). Inspired by this approach and our own recent results using [2-13C]-Glc as carbon sources for the simplification of ssNMR spectra, we present a strategy for a bi-directional sequential assignment of solid-state NMR resonances and additionally the detection of long-range contacts using the combination of 13C spin dilution and 3D NMR spectroscopy. We illustrate our results with the sequential assignment and the collection of distance restraints on an insoluble and non-crystalline supramolecular assembly, the Salmonella typhimurium type III secretion system needle.

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Language(s): eng - English
 Dates: 2012-12-012013
 Publication Status: Published in print
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 Rev. Method: Peer
 Identifiers: DOI: 10.1007/s10858-012-9691-9
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Title: Journal of biomolecular NMR
Source Genre: Journal
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Pages: - Volume / Issue: 55 (1) Sequence Number: - Start / End Page: 1 - 9 Identifier: ISSN: 0925-2738