Archaea-analoge Thiolipide für gestützte Lipiddoppelschichtmembranen auf ultraglatten Goldoberflächen

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…

Archaea Analogue Thiolipids for Tethered Bilayer Lipid Membranes on Ultrasmooth Gold Surfaces

Thiopeptide-Supported Lipid Layers on Solid Substrates

The sequential layer-by-layer formation of peptide-supported bimolecular lipid membranes at solid supports is described. In the first step, thiol-derivatized peptide sequences of 5 and 7 amino acids are assembled on a Au substrate. After activation of their COOH-terminus phospholipid molecules (DMPE) are covalently attached via an amid bond to form a tethered monolayer on the Au electrode. The different preparation steps are analyzed by Fourier transform IR, X-ray reflectometry, and surface plasmon spectroscopy. The latter technique is then also used to on-line monitor at the solid/solution interface the formation of a bilayer by fusion of vesicles prepared from a fluid lipid mixture with a…

Hydrogel-supported protein-tethered bilayer lipid membranes: a new approach toward polymer-supported lipid membranes

Polymer-supported bilayer lipid membranes offer great opportunities for the investigation of functional membrane proteins. Here we present a new approach in this direction by introducing a thin hydrogel layer as a soft ‘cushion’ on indium–tin oxide (ITO), providing a smooth, functional surface to form the protein-tethered BLM (ptBLM). ITO was used as a transparent electrode, enabling simultaneous implementation of electrochemical and optical waveguide techniques. The hydrogel poly(N-(2-hydroxyethyl)acrylamide-co-5-acrylamido-1-carboxypentyl-iminodiacetate-co-4-benzoylphenyl methacrylate) (P(HEAAm-co-NTAAAm-co-MABP)) was functionalized with the nickel chelating nitrilotriacetic acid (NTA) gr…

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.