Sensitive proton-detected solid-state NMR spectroscopy of large proteins with selective CH3 labelling: application to the 50S ribosome subunit
| Autor: | Paul Schanda, Pavel Macek, Vilius Kurauskas, Diego F. Gauto, Elodie Crublet, Rime Kerfah, Jérôme Boisbouvier |
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| Přispěvatelé: | Institut de biologie structurale (IBS - UMR 5075 ), Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS), European Research Council ERC-StG-2012-311318, European Research Council ERC-StG-2010-260887, Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA) |
| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Protein subunit 010402 general chemistry 01 natural sciences Ribosome Catalysis Article 03 medical and health sciences [CHIM.ANAL]Chemical Sciences/Analytical chemistry Labelling Materials Chemistry Spectroscopy 50S biology [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry Molecular Biology/Structural Biology [q-bio.BM] Chemistry Metals and Alloys General Chemistry Nuclear magnetic resonance spectroscopy Thermus thermophilus biology.organism_classification 0104 chemical sciences Surfaces Coatings and Films Electronic Optical and Magnetic Materials Crystallography 030104 developmental biology Solid-state nuclear magnetic resonance Ceramics and Composites Biophysics |
| Zdroj: | Chemical Communications Chemical Communications, Royal Society of Chemistry, 2016, 52, pp.9558-9561. ⟨10.1039/C6CC04484K⟩ Chemical Communications, 2016, 52, pp.9558-9561. ⟨10.1039/C6CC04484K⟩ |
| ISSN: | 1359-7345 1364-548X |
| DOI: | 10.1039/C6CC04484K⟩ |
| Popis: | International audience; Solid-state NMR spectroscopy allows the characterization of 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 only marginally increase. 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. |
| Databáze: | OpenAIRE |
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