title: A systematic mutagenesis-driven strategy for site-resolved NMR studies of supramolecular assemblies.
abstract: Obtaining sequence-specific assignments remains a major bottleneck in solution NMR investigations of supramolecular structure, dynamics and interactions. Here we demonstrate that resonance assignment of methyl probes in high molecular weight protein assemblies can be efficiently achieved by combining fast NMR experiments, residue-type-specific isotope-labeling and automated site-directed mutagenesis. The utility of this general and straightforward strategy is demonstrated through the characterization of intermolecular interactions involving a 468-kDa multimeric aminopeptidase, PhTET2.
authors.affiliation: Institut de Biologie Structurale Jean-Pierre Ebel, CNRS, Grenoble, France.
authors.forename: Carlos
authors.initials: C
authors.lastname: Amero
authors.forename: M
authors.initials: M
authors.lastname: Asunción Durá
authors.forename: Marjolaine
authors.initials: M
authors.lastname: Noirclerc-Savoye
authors.forename: Arnaud
authors.initials: A
authors.lastname: Perollier
authors.forename: Benoit
authors.initials: B
authors.lastname: Gallet
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Plevin
authors.forename: Thierry
authors.initials: T
authors.lastname: Vernet
authors.forename: Bruno
authors.initials: B
authors.lastname: Franzetti
authors.forename: Jérôme
authors.initials: J
authors.lastname: Boisbouvier
mesh_terms: D016297:Mutagenesis, Site-Directed / Q000379:methods*; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000737:chemistry* / Q000235:genetics*
chemical_list: D011506:Proteins