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Chem Commun (Camb). 2016 Jul 21;52(61):9558-61. doi: 10.1039/c6cc04484k.

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

Author information

1
Université Grenoble Alpes, Institut de Biologie Structurale, Grenoble, France. crublet@nmr-bio.com schanda@ibs.fr and CEA, Institut de Biologie Structurale, F-38044 Grenoble, France and CNRS, Institut de Biologie Structurale, F-38044 Grenoble, France.
2
NMR-Bio. 5 place Robert Schumann, F-38025 Grenoble, France.

Abstract

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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