caa a22 4500 445325798 CHVBK 20180317142720.0 cr unu---uuuuu 170323e20110701xx s 000 0 eng 10.1007/s10858-011-9513-5 doi (NATIONALLICENCE)springer-10.1007/s10858-011-9513-5 A systematic mutagenesis-driven strategy for site-resolved NMR studies of supramolecular assemblies [Elektronische Daten] [Carlos Amero, M. Asunción Durá, Marjolaine Noirclerc-Savoye, Arnaud Perollier, Benoit Gallet, Michael Plevin, Thierry Vernet, Bruno Franzetti, Jérôme Boisbouvier] 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. Springer Science+Business Media B.V., 2011 NMR spectroscopy nationallicence High molecular weight proteins nationallicence Assignment nationallicence Site-specific mutagenesis nationallicence Methyl-group labeling nationallicence Amero Carlos Institut de Biologie Structurale Jean-Pierre Ebel, CNRS, Grenoble, France aut Asunción Durá M. Institut de Biologie Structurale Jean-Pierre Ebel, CNRS, Grenoble, France aut Noirclerc-Savoye Marjolaine Institut de Biologie Structurale Jean-Pierre Ebel, CEA, Grenoble, France aut Perollier Arnaud Institut de Biologie Structurale Jean-Pierre Ebel, CNRS, Grenoble, France aut Gallet Benoit Institut de Biologie Structurale Jean-Pierre Ebel, CEA, Grenoble, France aut Plevin Michael Institut de Biologie Structurale Jean-Pierre Ebel, CNRS, Grenoble, France aut Vernet Thierry Institut de Biologie Structurale Jean-Pierre Ebel, CNRS, Grenoble, France aut Franzetti Bruno Institut de Biologie Structurale Jean-Pierre Ebel, CNRS, Grenoble, France aut Boisbouvier Jérôme Institut de Biologie Structurale Jean-Pierre Ebel, CNRS, Grenoble, France aut Journal of Biomolecular NMR Springer Netherlands 50/3(2011-07-01), 229-236 0925-2738 50:3<229 2011 50 10858 https://doi.org/10.1007/s10858-011-9513-5 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10858-011-9513-5 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Amero Carlos Institut de Biologie Structurale Jean-Pierre Ebel, CNRS, Grenoble, France aut NATIONALLICENCE 700 1- Asunción Durá M. Institut de Biologie Structurale Jean-Pierre Ebel, CNRS, Grenoble, France aut NATIONALLICENCE 700 1- Noirclerc-Savoye Marjolaine Institut de Biologie Structurale Jean-Pierre Ebel, CEA, Grenoble, France aut NATIONALLICENCE 700 1- Perollier Arnaud Institut de Biologie Structurale Jean-Pierre Ebel, CNRS, Grenoble, France aut NATIONALLICENCE 700 1- Gallet Benoit Institut de Biologie Structurale Jean-Pierre Ebel, CEA, Grenoble, France aut NATIONALLICENCE 700 1- Plevin Michael Institut de Biologie Structurale Jean-Pierre Ebel, CNRS, Grenoble, France aut NATIONALLICENCE 700 1- Vernet Thierry Institut de Biologie Structurale Jean-Pierre Ebel, CNRS, Grenoble, France aut NATIONALLICENCE 700 1- Franzetti Bruno Institut de Biologie Structurale Jean-Pierre Ebel, CNRS, Grenoble, France aut NATIONALLICENCE 700 1- Boisbouvier Jérôme Institut de Biologie Structurale Jean-Pierre Ebel, CNRS, Grenoble, France aut NATIONALLICENCE 773 0- Journal of Biomolecular NMR Springer Netherlands 50/3(2011-07-01), 229-236 0925-2738 50:3<229 2011 50 10858 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer