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

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

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Stoffregen, Mira C. Schwer, Matthias M. Renschler, Fabian A. Wiesner, Silke
Description	Country affiliation: Germany Author Affiliation: Stoffregen MC ( Max Planck Institute for Developmental Biology, Spemannstrasse 35, 72076 Tübingen, Germany.)
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.
File Format	HTM / HTML
ISSN	09692126
Issue Number	4
Volume Number	20
e-ISSN	18784186
Journal	Structure
Language	English
Publisher	Elsevier
Publisher Date	2012-04-04
Publisher Place	United States
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Discipline Molecular Biology Discipline Biochemistry Discipline Biotechnology Endosomal Sorting Complexes Required For Transport Chemistry Methionine Saccharomyces Cerevisiae Proteins Saccharomyces Cerevisiae Metabolism Ubiquitin-conjugating Enzymes Ubiquitin-protein Ligase Complexes Binding Sites Genetics Escherichia Coli Models, Molecular Mutation Nuclear Magnetic Resonance, Biomolecular Plasmids Protein Binding Protein Conformation Protein Isoforms Protein Structure, Tertiary Recombinant Proteins Transfection Journal Article Research Support, Non-u.s. Gov't
Content Type	Text
Resource Type	Article
Subject	Structural Biology Molecular Biology

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