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@ARTICLE{Mckel:872747,
author = {Möckel, Christina and Kubiak, Jakub and Schillinger,
Oliver and Kühnemuth, Ralf and Della Corte, Dennis and
Schröder, Gunnar F. and Willbold, Dieter and Strodel,
Birgit and Seidel, Claus A. M. and Neudecker, Philipp},
title = {{I}ntegrated {NMR}, {F}luorescence, and {M}olecular
{D}ynamics {B}enchmark {S}tudy of {P}rotein {M}echanics and
{H}ydrodynamics},
journal = {The journal of physical chemistry / B B, Condensed matter,
materials, surfaces, interfaces $\&$ biophysical},
volume = {123},
number = {7},
issn = {1520-5207},
address = {Washington, DC},
publisher = {Soc.},
reportid = {FZJ-2020-00225},
pages = {1453 - 1480},
year = {2019},
abstract = {Understanding the function of a protein requires not only
knowledge of its tertiary structure but also an
understanding of its conformational dynamics. Nuclear
magnetic resonance (NMR) spectroscopy, polarization-resolved
fluorescence spectroscopy and molecular dynamics (MD)
simulations are powerful methods to provide detailed insight
into protein dynamics on multiple time scales by monitoring
global rotational diffusion and local flexibility (order
parameters) that are sensitive to inter- and intramolecular
interactions, respectively. We present an integrated
approach where data from these techniques are analyzed and
interpreted within a joint theoretical description of
depolarization and diffusion, demonstrating their conceptual
similarities. This integrated approach is then applied to
the autophagy-related protein GABARAP in its cytosolic form,
elucidating its dynamics on the pico- to nanosecond time
scale and its rotational and translational diffusion for
protein concentrations spanning 9 orders of magnitude. We
compare the dynamics of GABARAP as monitored by 15N spin
relaxation of the backbone amide groups, fluorescence
anisotropy decays and fluorescence correlation spectroscopy
of side chains labeled with BODIPY FL, and molecular movies
of the protein from MD simulations. The recovered parameters
agree very well between the distinct techniques if the
different measurement conditions (probe localization, sample
concentration) are taken into account. Moreover, we propose
a method that compares the order parameters of the backbone
and side chains to identify potential hinges for
large-scale, functionally relevant intradomain motions, such
as residues 27/28 at the interface between the two
subdomains of GABARAP. In conclusion, the integrated concept
of cross-fertilizing techniques presented here is
fundamental to obtaining a comprehensive quantitative
picture of multiscale protein dynamics and solvation. The
possibility to employ these validated techniques under
cellular conditions and combine them with fluorescence
imaging opens up the perspective of studying the functional
dynamics of GABARAP or other proteins in live cells.},
cin = {ICS-6 / JARA-HPC},
ddc = {530},
cid = {I:(DE-Juel1)ICS-6-20110106 / $I:(DE-82)080012_20140620$},
pnm = {551 - Functional Macromolecules and Complexes (POF3-551) /
IHRS-BioSoft - International Helmholtz Research School of
Biophysics and Soft Matter (IHRS-BioSoft-20061101) /
Structural dynamics of murine guanylate binding proteins,
their dimerization and interaction with lipid bilayers
$(jics6a_20180501)$},
pid = {G:(DE-HGF)POF3-551 / G:(DE-Juel1)IHRS-BioSoft-20061101 /
$G:(DE-Juel1)jics6a_20180501$},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:30525615},
UT = {WOS:000459836700001},
doi = {10.1021/acs.jpcb.8b08903},
url = {https://juser.fz-juelich.de/record/872747},
}