Incorporation of Membrane Proteins in Solid-Supported Lipid Layers

Incorporation of the acetylcholine receptor dimer from Torpedo californica in a peptide supported lipid membrane investigated by surface plasmon and fluorescence spectroscopy

Abstract The dimer species (Mr 580 000) of the nicotinic acetylcholine receptor, isolated from the electric organ of Torpedo californica, was incorporated into a thiopeptide supported lipid bilayer. The incorporation was achieved by fusion of liposomes with reconstituted receptor onto a gold-supported thiopeptide lipid monolayer. Surface plasmon resonance spectroscopy (SPS) was used to monitor in real time the fusion process as well as the specific binding of the antagonist α-bungarotoxin. A recently developed extension of SPS offering enhanced sensitivity and specificity, surface plasmon fluorescence spectroscopy (SPFS), was then used to monitor subsequent binding of the monoclonal WF6 and…

Inkorporation von Membranproteinen in festkörperunterstützte planare Lipidfilme

Coupling of proton translocation through ATPase incorporated into supported lipid bilayers to an electrochemical process

Abstract H+-ATPase is incorporated into solid-supported lipid bilayers separated from the gold support by a peptide spacer. The translocation of protons across the lipid film to the inner side is coupled to the discharge of protons at the gold surface. The overall process is investigated by square wave voltammetry (SWV) and double potential-pulse chronoamperometry (CA). As a result, the formation of a proton gradient is monitored by SWV whereas currents measured by CA monitor the stationary state when the enzyme activity is directly coupled to the charge transfer at the electrode. These currents markedly depend on the number of ATPases present in the bilayer.