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@ARTICLE{Lakomek:1031670,
author = {Lakomek, Nils and Stief, Tobias and Vormann, Katharina},
title = {{NMR} 15{N} {R}elaxation {E}xperiments for the
{I}nvestigation of {P}icosecond to {N}anoseconds
{S}tructural {D}ynamics of {P}roteins},
journal = {JoVE journal},
volume = {},
issn = {1940-087X},
address = {Cambridge, MA},
publisher = {JoVE},
reportid = {FZJ-2024-05780},
pages = {e67088},
year = {2024},
abstract = {Nuclear magnetic resonance (NMR) spectroscopy allows
studying proteins in solution and under physiological
temperatures. Frequently, either the amide groups of the
protein backbone or the methyl groups in side chains are
used as reporters of structural dynamics in proteins. A
structural dynamics study of the protein backbone of
globular proteins on 15N labeled and fully protonated
samples usually works well for proteins with a molecular
weight of up to 50 kDa. When side chain deuteration in
combination with transverse relaxation optimized
spectroscopy (TROSY) is applied, this limit can be extended
up to 200 kDa for globular proteins and up to 1 MDa when the
focus is on the side chains. When intrinsically disordered
proteins (IDPs) or proteins with intrinsically disordered
regions (IDRs) are investigated, these weight limitations do
not apply but can go well beyond. The reason is that IDPs or
IDRs, characterized by high internal flexibility, are
frequently dynamically decoupled. Various NMR methods offer
atomic-resolution insights into structural protein dynamics
across a wide range of time scales, from picoseconds up to
hours. Standard 15N relaxation measurements overview a
protein's internal flexibility and characterize the protein
backbone dynamics experienced on the fast pico- to
nanosecond timescale. This article presents a hands-on
protocol for setting up and recording NMR 15N R1, R2, and
heteronuclear Overhauser effect (hetNOE) experiments. We
show exemplary data and explain how to interpret them simply
qualitatively before any more sophisticated analysis.},
cin = {IBI-7},
ddc = {570},
cid = {I:(DE-Juel1)IBI-7-20200312},
pnm = {5241 - Molecular Information Processing in Cellular Systems
(POF4-524)},
pid = {G:(DE-HGF)POF4-5241},
typ = {PUB:(DE-HGF)16},
pubmed = {39555784},
UT = {WOS:001368152600029},
doi = {10.3791/67088},
url = {https://juser.fz-juelich.de/record/1031670},
}