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@ARTICLE{Kotov:435901,
author = {Kotov, Vadim and Bartels, Kim and Veith, Katharina and
Josts, Inokentijs and Subhramanyam, Udaya K. Tiruttani and
Günther, Christian and Labahn, Jörg and Marlovits, Thomas
C. and Moraes, Isabel and Tidow, Henning and Loew, Christian
and Garcia-Alai, Maria M.},
title = {{H}igh-throughput stability screening for
detergent-solubilized membrane proteins},
journal = {Scientific reports},
volume = {9},
number = {1},
issn = {2045-2322},
address = {[London]},
publisher = {Macmillan Publishers Limited, part of Springer Nature},
reportid = {PUBDB-2020-00849},
pages = {10379},
year = {2019},
abstract = {Protein stability in detergent or membrane-like
environments is the bottleneck for structural studies on
integral membrane proteins (IMP). Irrespective of the method
to study the structure of an IMP, detergent solubilization
from the membrane is usually the first step in the workflow.
Here, we establish a simple, high-throughput screening
method to identify optimal detergent conditions for membrane
protein stabilization. We apply differential scanning
fluorimetry in combination with scattering upon thermal
denaturation to study the unfolding of integral membrane
proteins. Nine different prokaryotic and eukaryotic membrane
proteins were used as test cases to benchmark our detergent
screening method. Our results show that it is possible to
measure the stability and solubility of IMPs by diluting
them from their initial solubilization condition into
different detergents. We were able to identify groups of
detergents with characteristic stabilization and
destabilization effects for selected targets. We further
show that fos-choline and PEG family detergents may lead to
membrane protein destabilization and unfolding. Finally, we
determined thenmodynamic parameters that are important
indicators of IMP stability. The described protocol allows
the identification of conditions that are suitable for
downstream handling of membrane proteins during
purification.},
cin = {CSSB-UKE / CSSB-EMBL-CL / CSSB-UKE-TM / CSSB-FZJ-JL /
CSSB-CF-SPC},
ddc = {600},
cid = {I:(DE-H253)CSSB-UKE-20141216 /
I:(DE-H253)CSSB-EMBL-CL-20210806 /
I:(DE-H253)CSSB-UKE-TM-20210520 /
I:(DE-H253)CSSB-FZJ-JL-20210520 /
I:(DE-H253)CSSB-CF-SPC-20210520},
pnm = {6215 - Soft Matter, Health and Life Sciences (POF3-621)},
pid = {G:(DE-HGF)POF3-6215},
experiment = {EXP:(DE-MLZ)NOSPEC-20140101},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:31316088},
UT = {WOS:000475832700023},
doi = {10.1038/s41598-019-46686-8},
url = {https://bib-pubdb1.desy.de/record/435901},
}