TY  - JOUR
AU  - Kotov, Vadim
AU  - Bartels, Kim
AU  - Veith, Katharina
AU  - Josts, Inokentijs
AU  - Subhramanyam, Udaya K. Tiruttani
AU  - Günther, Christian
AU  - Labahn, Jörg
AU  - Marlovits, Thomas C.
AU  - Moraes, Isabel
AU  - Tidow, Henning
AU  - Loew, Christian
AU  - Garcia-Alai, Maria M.
TI  - High-throughput stability screening for detergent-solubilized membrane proteins
JO  - Scientific reports
VL  - 9
IS  - 1
SN  - 2045-2322
CY  - [London]
PB  - Macmillan Publishers Limited, part of Springer Nature
M1  - PUBDB-2020-00849
SP  - 10379
PY  - 2019
AB  - 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.
LB  - PUB:(DE-HGF)16
C6  - pmid:31316088
UR  - <Go to ISI:>//WOS:000475832700023
DO  - DOI:10.1038/s41598-019-46686-8
UR  - https://bib-pubdb1.desy.de/record/435901
ER  -