Biomimetic tethered lipid membranes designed for membrane-protein interaction studies

Abstract

The complexity of the biological membranes restricts their direct investigation at the nanoscale. Lipid bilayer membranes have been developed as a model of biological membranes in order to allow the interaction and insertion of peptides and membrane proteins in a functional manner. Promising models have been developed in the past two decades and tethered bilayer design traduces constant improvement of membrane models. The formation of protein free solid tethered membranes can be achieved by direct vesicle fusion, Langmuir-Blodgett, Langmuir-Schaffer transfers, self assembly of various building blocks such as thiol on gold, silane on quartz, grafting of polymers, as well as ligand receptor recognition. In this review, the current state of different tethered bilayer membrane will be described. We will focus on critical analysis of the main advantages/drawbacks of each kind of model construction and their ability to allow protein incorporation in non-denaturing conditions. Some of the current drawbacks encountered in these biomimetic models can be overcome using an innovative tethered bilayer design based on a reliable and fast formation method. The successful protein incorporation of the Adenylate Cyclase produced by Bordetella pertussis and the voltage dependent anion channel (VDAC) was demonstrated on this model.

Bordetella pertussis adenylate cyclase toxin translocation across a tethered lipid bilayer

“…2B; Table S1, experiment B; and Fig. S2B), a value consistent with the formation of a planar lipid bilayer, as reported previously (40,42). The fluidity and continuity of the bilayer were confirmed by fluorescence recovery after photobleaching experiments, as described in our previous report (40).…”

“…These biomimetic models have been widely used to characterize protein-membrane associations, binding of ligands to cellular receptors incorporated in the supported planar bilayers, and molecular interactions in cell adhesion processes (41,42,52,53); however, up to now, few studies have been devoted to protein transport across reconstituted bilayers, owing to various technical difficulties. Our tBLM/CaM technique is particularly well adapted for characterizing protein translocation across membranes, allowing for direct detection of the passage of polypeptides from the cis side to the trans side of the membrane.…”

Functional liposomes and supported lipid bilayers: towards the complexity of biological archetypes

“…In parallel, a multitude of more ambitious architectures have been proposed during the past decade spanning from hybrid bilayers involving vesicle fusion combined with LB or SAM approaches 25 to tethered lipid bilayers, 26,27 suspended lipid bilayers or supported vesicular layers. Nevertheless, vesicle fusion on bare substrates is certainly the most extensively studied procedure, although the quest for a mimetic model of intracellular crowding certainly invites the adoption of a cushioned SLB.…”

“…27 Other strategies may be more fruitful to develop stable and reliable asymmetrical SLB: these methods have been detailed in recent reports 27 and include the deposition of a LB layer on a hydrophilic solid support followed by adsorption and disruption of vesicle with a different phospholipid composition compared to the first LB layer (LB/VF method). This procedure, although simple, results in asymmetrical bilayers whose fluidity properties closely match those of the native LB layers.…”

Application of Polarization Modulation Infrared Reflection Absorption Spectrosocpy Under Electrochemical Control for Structural Studies of Biomimetic Assemblies

“…In order to approach this multidisciplinary and complex problem connecting surface chemistry with biology and material science advanced physicochemical and analytical techniques have to be used [8][9][10]. The deposition of biomimetic films on the electrode surfaces allows studies of the electric-field driven changes in these assemblies [9,11].…”