title: Surface-supported bilayers with transmembrane proteins: role of the polymer cushion revisited.
abstract: Protein lateral mobility in surface-supported bilayers is often much lower than the mobility of the lipids. In the present study we explore whether the incorporation of a PEG cushion between the bilayer and the substrate increases the lateral mobility of transmembrane proteins in bilayers produced via directed assembly, a method based on Langmuir-Blodgett deposition techniques. In our experiments, the PEG cushions were incorporated by adding PEG lipids to the protein/lipid monolayer at the air/water interface, at the first step of bilayer assembly. The protein and lipid mobilities in 160 different bilayers, with various PEG molecular weights and PEG lipid concentrations, were measured and compared. We found that the measured diffusion coefficients do not depend on the PEG molecular weight or the PEG lipid concentration and are very similar to the values measured in the absence of PEG. Therefore, contrary to our expectations, we found that a PEG cushion does not necessarily increase protein mobility, suggesting that the low protein mobility is not a consequence of protein-substrate interactions. Furthermore, we showed that the low protein mobility is not due to protein aggregation. The major determinant of protein mobility in surface-supported bilayer systems appears to be the method of bilayer assembly. While proteins were always mobile if the bilayers were prepared using the directed assembly method, in the presence and absence of a PEG cushion, other bilayer assembly protocols resulted in complete lack of protein mobility.
authors.affiliation: Department of Materials Science and Engineering, Johns Hopkins University, Baltimore, Maryland 21218, USA.
authors.forename: Mikhail
authors.initials: M
authors.lastname: Merzlyakov
authors.forename: Edwin
authors.initials: E
authors.lastname: Li
authors.forename: Ivan
authors.initials: I
authors.lastname: Gitsov
authors.forename: Kalina
authors.initials: K
authors.lastname: Hristova
mesh_terms: D000327:Adsorption; D000388:Air; D002621:Chemistry / Q000379:methods; D031541:Fluorescence Resonance Energy Transfer; D008051:Lipid Bilayers / Q000737:chemistry*; D008055:Lipids / Q000737:chemistry; D008565:Membrane Proteins / Q000737:chemistry*; D008956:Models, Chemical; D008968:Molecular Conformation; D010455:Peptides / Q000737:chemistry; D011092:Polyethylene Glycols / Q000737:chemistry*; D011108:Polymers / Q000737:chemistry*; D013379:Substrate Specificity; D013499:Surface Properties
chemical_list: D008051:Lipid Bilayers; D008055:Lipids; D008565:Membrane Proteins; D010455:Peptides; D011108:Polymers; D011092:Polyethylene Glycols