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

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 combinin...

Full description

Saved in:
Bibliographic Details
Published in:Journal of biomolecular NMR 2011-07, Vol.50 (3), p.229-236
Main Authors: Amero, Carlos, Asunción Durá, M., Noirclerc-Savoye, Marjolaine, Perollier, Arnaud, Gallet, Benoit, Plevin, Michael J., Vernet, Thierry, Franzetti, Bruno, Boisbouvier, Jérôme
Format: Article
Language:English
Subjects:
Biochemistry
Biological and Medical Physics
Biophysics
Mutagenesis, Site-Directed - methods
Nuclear Magnetic Resonance, Biomolecular - methods
Physics
Physics and Astronomy
Protein Structure, Secondary
Protein Structure, Tertiary
Proteins - chemistry
Proteins - genetics
Spectroscopy/Spectrometry
Citations: Items that this one cites
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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.
ISSN:0925-2738
1573-5001
DOI:10.1007/s10858-011-9513-5