title: Methyl-specific isotopic labeling: a molecular tool box for solution NMR studies of large proteins.
abstract: Nuclear magnetic resonance (NMR) spectroscopy is a uniquely powerful tool for studying the structure, dynamics and interactions of biomolecules at atomic resolution. In the past 15 years, the development of new isotopic labeling strategies has opened the possibility of exploiting NMR spectroscopy in the study of supra-molecular complexes with molecular weights of up to 1MDa. At the core of these isotopic labeling developments is the specific introduction of [(1)H,(13)C]-labeled methyl probes into perdeuterated proteins. Here, we describe the evolution of these approaches and discuss their impact on structural and biological studies. The relevant protocols are succinctly reviewed for single and combinatorial isotopic-labeling of methyl-containing residues, and examples of applications on challenging biological systems, including high molecular weight and membrane proteins, are presented.
authors.affiliation: University Grenoble Alpes, Institut de Biologie Structurale (IBS), F-38044 Grenoble, France; CNRS, F-38044 Grenoble, France; CEA, DSV, F-38044 Grenoble, France.
authors.forename: Rime
authors.initials: R
authors.lastname: Kerfah
authors.affiliation: Department of Biology, University of York, York YO10 5DD, UK.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Plevin
authors.affiliation: Institut de Génomique Fonctionnelle - CNRS - INSERM - University Montpellier I & II, F-34094 Montpellier, France.
authors.forename: Remy
authors.initials: R
authors.lastname: Sounier
authors.affiliation: University Grenoble Alpes, Institut de Biologie Structurale (IBS), F-38044 Grenoble, France; CNRS, F-38044 Grenoble, France; CEA, DSV, F-38044 Grenoble, France.
authors.forename: Pierre
authors.initials: P
authors.lastname: Gans
authors.affiliation: University Grenoble Alpes, Institut de Biologie Structurale (IBS), F-38044 Grenoble, France; CNRS, F-38044 Grenoble, France; CEA, DSV, F-38044 Grenoble, France. Electronic address: jerome.boisbouvier@ibs.fr.
authors.forename: Jerome
authors.initials: J
authors.lastname: Boisbouvier
mesh_terms: D000818:Animals; D002247:Carbon Isotopes / Q000737:chemistry; D003903:Deuterium / Q000737:chemistry; D006801:Humans; D007553:Isotope Labeling / Q000379:methods; D008565:Membrane Proteins / Q000737:chemistry; D008745:Methylation; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry*
chemical_list: D002247:Carbon Isotopes; D008565:Membrane Proteins; D011506:Proteins; D003903:Deuterium