Insights into co-translational membrane protein insertion by combined LILBID-mass spectrometry and NMR spectroscopy.
Related Articles Insights into co-translational membrane protein insertion by combined LILBID-mass spectrometry and NMR spectroscopy.
Anal Chem. 2017 Oct 17;:
Authors: Peetz O, Henrich E, Laguerre A, L?hr F, Hein C, Dötsch V, Bernhard F, Morgner N
Abstract
Co-translational insertion of membrane proteins into defined nanoparticle membranes has been developed as an efficient process to produce highly soluble samples in native-like environments and to study lipid dependent effects on protein structure and function. Numerous examples of structural and functional characterization of transporters, ion channels or G-protein coupled receptors in co-translationally formed nanodisc complexes demonstrate the versatility of this approach, although the basic underlying mechanisms of membrane insertion are mainly unknown. We have revealed first aspects of the insertion of proteins into nanodiscs by combining cell-free expression, non-covalent mass spectrometry and NMR spectroscopy. We provide evidence of cooperative insertion of homooligomeric complexes and demonstrate the possibility to modulate their stoichiometry by modifying reaction conditions. Additionally, we show that significant amounts of lipid are released from the nanodiscs upon insertion of larger protein complexes.
PMID: 29039652 [PubMed - as supplied by publisher]
More...