title: Sensitive proton-detected solid-state NMR spectroscopy of large proteins with selective CH3 labelling: application to the 50S ribosome subunit.
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.
authors.affiliation: 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.
authors.forename: Vilius
authors.initials: V
authors.lastname: Kurauskas
authors.affiliation: 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.
authors.forename: Elodie
authors.initials: E
authors.lastname: Crublet
authors.affiliation: 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.
authors.forename: Pavel
authors.initials: P
authors.lastname: Macek
authors.affiliation: NMR-Bio. 5 place Robert Schumann, F-38025 Grenoble, France.
authors.forename: Rime
authors.initials: R
authors.lastname: Kerfah
authors.affiliation: 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.
authors.forename: Diego F
authors.initials: DF
authors.lastname: Gauto
authors.affiliation: 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.
authors.forename: Jérôme
authors.initials: J
authors.lastname: Boisbouvier
authors.affiliation: 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.
authors.forename: Paul
authors.initials: P
authors.lastname: Schanda