TY  - JOUR
A1  - Henrich, Erik
A1  - Peetz, Oliver
A1  - Hein, Christopher
A1  - Laguerre, Aisha
A1  - Hoffmann, Beate
A1  - Hoffmann, Jan
A1  - Dötsch, Volker
A1  - Bernhard, Frank
A1  - Morgner, Nina
T1  - Analyzing native membrane protein assembly in nanodiscs by combined non-covalent mass spectrometry and synthetic biology
T2  - eLife
N2  - Membrane proteins frequently assemble into higher order homo- or hetero-oligomers within their natural lipid environment. This complex formation can modulate their folding, activity as well as substrate selectivity. Non-disruptive methods avoiding critical steps, such as membrane disintegration, transfer into artificial environments or chemical modifications are therefore essential to analyze molecular mechanisms of native membrane protein assemblies. The combination of cell-free synthetic biology, nanodisc-technology and non-covalent mass spectrometry provides excellent synergies for the analysis of membrane protein oligomerization within defined membranes. We exemplify our strategy by oligomeric state characterization of various membrane proteins including ion channels, transporters and membrane-integrated enzymes assembling up to hexameric complexes. We further indicate a lipid-dependent dimer formation of MraY translocase correlating with the enzymatic activity. The detergent-free synthesis of membrane protein/nanodisc samples and the analysis by LILBID mass spectrometry provide a versatile platform for the analysis of membrane proteins in a native environment.
KW  - Tools and resources
KW  - Biochemistry
KW  - Biophysics and structural biology
KW  - Cell-free synthetic biology
KW  - Non-covalent mass spectrometry
KW  - Nanodisc
KW  - Membrane protein
KW  - Oligomeric State
Y1  - 2017
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/45646
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-456463
SN  - 2050-084X
N1  - Copyright Henrich et al. This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.
VL  - 6
IS  - e20954
SP  - 1
EP  - 19
PB  - eLife Sciences Publications
CY  - Cambridge
ER  -