English
 
Help Privacy Policy Disclaimer
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT

Released

Journal Article

Methionine scanning as an NMR tool for detecting and analyzing biomolecular interaction surfaces

MPS-Authors
/persons/resource/persons273278

Stoffregen,  MC
Research Group Mechanisms of Ubiquitin-dependent Cell Signaling, Max Planck Institute for Developmental Biology, Max Planck Society;

/persons/resource/persons275818

Schwer,  MM
Research Group Mechanisms of Ubiquitin-dependent Cell Signaling, Max Planck Institute for Developmental Biology, Max Planck Society;

/persons/resource/persons273872

Renschler,  FA
Research Group Mechanisms of Ubiquitin-dependent Cell Signaling, Max Planck Institute for Developmental Biology, Max Planck Society;

/persons/resource/persons273275

Wiesner,  S
Research Group Mechanisms of Ubiquitin-dependent Cell Signaling, Max Planck Institute for Developmental Biology, Max Planck Society;

External Resource
No external resources are shared
Fulltext (public)
There are no public fulltexts stored in PuRe
Supplementary Material (public)
There is no public supplementary material available
Citation

Stoffregen, M., Schwer, M., Renschler, F., & Wiesner, S. (2012). Methionine scanning as an NMR tool for detecting and analyzing biomolecular interaction surfaces. Structure, 20(4), 573-581. doi:10.1016/j.str.2012.02.012.


Cite as: http://hdl.handle.net/21.11116/0000-000A-AFBF-B
Abstract
Methyl NMR spectroscopy is a powerful tool for studying protein structure, dynamics, and interactions. Yet difficulties with resonance assignment and the low abundance of methyl groups can preclude detailed NMR studies, particularly the determination of continuous interaction surfaces. Here we present a straightforward strategy that overcomes these problems. We systematically substituted solvent-exposed residues with reporter methionines in the expected binding site and performed chemical shift perturbation (CSP) experiments using methyl-TROSY spectra. We demonstrate the utility of this approach for the interaction between the HECT domain of the Rsp5p ubiquitin ligase and its cognate E2, Ubc4. Using these mutants, we could instantaneously assign all newly arising reporter methyl signals, determine the Ubc4 interaction surface on a per-residue basis, and investigate the importance of each individual mutation for ligand binding. Our data show that methionine scanning significantly extends the applicability, information content, and spatial resolution of methyl CSP experiments.