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  Proton-detected MAS NMR experiments based on dipolar transfers for backbone assignment of highly deuterated proteins.

Chevelkov, V., Habenstein, B., Loquet, A., Giller, K., Becker, S., & Lange, A. (2014). Proton-detected MAS NMR experiments based on dipolar transfers for backbone assignment of highly deuterated proteins. Journal of Magnetic Resonance, 242, 180-188. doi:10.1016/j.jmr.2014.02.020.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0019-8BC4-9 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-002A-096D-3
Genre: Journal Article

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 Creators:
Chevelkov, V.1, Author              
Habenstein, B.1, Author              
Loquet, A.1, Author              
Giller, K.2, Author              
Becker, S.2, Author              
Lange, A.2, 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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 Abstract: Proton-detected solid-state NMR was applied to a highly deuterated insoluble, non-crystalline biological assembly, the Salmonella typhimurium type iii secretion system (T3SS) needle. Spectra of very high resolution and sensitivity were obtained at a low protonation level of 10-20% at exchangeable amide positions. We developed efficient experimental protocols for resonance assignment tailored for this system and the employed experimental conditions. Using exclusively dipolar-based interspin magnetization transfers, we recorded two sets of 3D spectra allowing for an almost complete backbone resonance assignment of the needle subunit PrgI. The additional information provided by the well-resolved proton dimension revealed the presence of two sets of resonances in the N-terminal helix of PrgI, while in previous studies employing 13C detection only a single set of resonances was observed.

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Language(s): eng - English
 Dates: 2014-03-042014-05
 Publication Status: Published in print
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 Rev. Method: Peer
 Identifiers: DOI: 10.1016/j.jmr.2014.02.020
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Title: Journal of Magnetic Resonance
Source Genre: Journal
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Pages: - Volume / Issue: 242 Sequence Number: - Start / End Page: 180 - 188 Identifier: -