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Transfer on hydrophobic substrates and AFM imaging of membrane proteins reconstituted in planar lipid bilayers
Author(s) -
Seantier Bastien,
Dezi Manuela,
Gubellini Francesca,
Berquand Alexandre,
Godefroy Cédric,
Dosset Patrice,
Lévy Daniel,
Milhiet PierreEmmanuel
Publication year - 2011
Publication title -
journal of molecular recognition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.401
H-Index - 79
eISSN - 1099-1352
pISSN - 0952-3499
DOI - 10.1002/jmr.1070
Subject(s) - monolayer , membrane , lipid bilayer , model lipid bilayer , chemistry , highly oriented pyrolytic graphite , biological membrane , mica , nanotechnology , biophysics , crystallography , materials science , lipid bilayer phase behavior , graphite , organic chemistry , biochemistry , composite material , biology
The lipid‐layer technique allows reconstituting transmembrane proteins at a high density in microns size planar membranes and suspended to a lipid monolayer at the air/water interface. In this paper, we transferred these membranes onto two hydrophobic substrates for further structural analysis of reconstituted proteins by Atomic Force Microscopy (AFM). We used a mica sheet covered by a lipid monolayer or a sheet of highly oriented pyrolytic graphite (HOPG) to trap the lipid monolayer at the interface and the suspended membranes. In both cases, we succeeded in the transfer of large membrane patches containing densely packed or 2D‐crystallized proteins. As a proof of concept, we transferred and imaged the soluble Shiga toxin bound to its lipid ligand and the ATP‐binding cassette (ABC) transporter BmrA reconstituted into a planar bilayer. AFM imaging with a lateral resolution in the nanometer range was achieved. Potential applications of this technique in structural biology and nanobiotechnology are discussed. Copyright © 2011 John Wiley & Sons, Ltd.