Arginine Dynamics Probed by Magic-Angle Spinning NMR with a Specific Isotope-Labeling Scheme

DOI

The specific introduction of 1H-13C or 1H-15N moieties into otherwise deuterated proteins holds great potential for high-resolution solution and magic-angle spinning (MAS) NMR studies of protein structure and dynamics. Arginine residues play key roles for example at active sites of enzymes. Taking advantage of a chemically synthesized Arg with a 13C-1H2 group in an otherwise deuterated backbone, we demonstrate here the usefulness of proton-detected arginine MAS NMR approaches to probe arginine dynamics. In experiments on crystalline ubiquitin and the 134 kDa tetrameric enzyme malate dehydrogenase we detected a wide range of motions, from sites that are rigid on time scales of at least tens of milliseconds to residues undergoing predominantly nanosecond motions. Spin-relaxation and dipolar-coupling measurements enabled quantitative determination of these dynamics. We observed microsecond dynamics of residue Arg54 in crystalline ubiquitin, whose backbone is known to sample different β-turn conformations on this time scale. The labeling scheme and experiments presented here expand the toolkit for high-resolution proton-detected MAS NMR

Identifier
DOI https://doi.org/10.15479/AT-ISTA-19956
Metadata Access https://research-explorer.app.ist.ac.at/oai?verb=GetRecord&metadataPrefix=oai_dc&identifier=oai:pub.research-explorer.ista.ac.at:19956
Provenance
Creator Schanda, Paul ; ORCID logo
Publisher Institute of Science and Technology Austria
Publication Year 2025
Rights info:eu-repo/semantics/openAccess
OpenAccess true
Contact repository.manager(at)ist.ac.at
Representation
Resource Type info:eu-repo/semantics/other; doc-type:ResearchData; Text
Discipline Life Sciences, Natural Sciences, Engineering Sciences