A systematic mutagenesis-driven strategy for site-resolved NMR studies of supramolecular assemblies
 

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.

• Content Type Journal Article
• Pages 1-8
• DOI 10.1007/s10858-011-9513-5
• Authors
  • Carlos Amero, Institut de Biologie Structurale Jean-Pierre Ebel, CNRS, Grenoble, France
  • M. Asunción Durá, Institut de Biologie Structurale Jean-Pierre Ebel, CNRS, Grenoble, France
  • Marjolaine Noirclerc-Savoye, Institut de Biologie Structurale Jean-Pierre Ebel, CEA, Grenoble, France
  • Arnaud Perollier, Institut de Biologie Structurale Jean-Pierre Ebel, CNRS, Grenoble, France
  • Benoit Gallet, Institut de Biologie Structurale Jean-Pierre Ebel, CEA, Grenoble, France
  • Michael J. Plevin, Institut de Biologie Structurale Jean-Pierre Ebel, CNRS, Grenoble, France
  • Thierry Vernet, Institut de Biologie Structurale Jean-Pierre Ebel, CNRS, Grenoble, France
  • Bruno Franzetti, Institut de Biologie Structurale Jean-Pierre Ebel, CNRS, Grenoble, France
  • Jérôme Boisbouvier, Institut de Biologie Structurale Jean-Pierre Ebel, CNRS, Grenoble, France

• • Journal Journal of Biomolecular NMR
  • Online ISSN 1573-5001
  • Print ISSN 0925-2738

Source: Journal of Biomolecular NMR