Sensitive proton-detected solid-state NMR spectroscopy of large proteins with selective CH3 labelling: application to the 50S ribosome subunit

Solid-state NMR spectroscopy allows the characterization of the structure, interactions and dynamics of insoluble and/or very large proteins. Sensitivity and resolution are often major challenges for obtaining atomic-resolution information, in particular for very large protein complexes. Here we sho...

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Published inChemical communications (Cambridge, England) Vol. 52; no. 61; pp. 9558 - 9561
Main Authors Kurauskas, Vilius, Crublet, Elodie, Macek, Pavel, Kerfah, Rime, Gauto, Diego F, Boisbouvier, Jérôme, Schanda, Paul
Format Journal Article
LanguageEnglish
Published England Royal Society of Chemistry 21.07.2016
Subjects
Analytical chemistry
Biochemistry, Molecular Biology
Chemical Sciences
Deuteration
Dynamics
Gain
Labelling
Life Sciences
NMR spectroscopy
Proteins
Strategy
Structural analysis
Structural Biology
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Summary:Solid-state NMR spectroscopy allows the characterization of the structure, interactions and dynamics of insoluble and/or very large proteins. Sensitivity and resolution are often major challenges for obtaining atomic-resolution information, in particular for very large protein complexes. Here we show that the use of deuterated, specifically CH3-labelled proteins result in significant sensitivity gains compared to previously employed CHD2 labelling, while line widths increase only marginally. We apply this labelling strategy to a 468 kDa-large dodecameric aminopeptidase, TET2, and the 1.6 MDa-large 50S ribosome subunit of Thermus thermophilus.
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Present address: NMR-Bio. 5 place Robert Schumann, F-38044 Grenoble, France
ISSN:1359-7345
1364-548X
DOI:10.1039/c6cc04484k