Surface-bound biomembranes incorporating receptors: electrochemical and structural characterization

A generic method is described for forming surface-bound structures that incorporate protein receptors in a membrane-like environment. Silane reagents (octadecyltrichlorosilane and dimethyloctadecylchlorosilane) were used to produce primed substrates bearing full and partial monolayers, respectively. Biomembranes were formed by dialysis of detergent-solubilized membranes in the presence of two different alkylsilanized substrates: Si/SiO2 electrodes and glass microspheres. Electrochemical analysis of the capacitance was used to determine apparent thickness and degree of surface coverage at each stage in the deposition process. Elemental analysis on glass beads gave the hydrocarbon incorporation. Glass bead substrates were also examined by Fourier transform infrared spectroscopy to evaluate the alkylsilanized substrate before and after dialysis. Both vertebrate rhodopsin and the nicotinic acetylcholine receptor could be incorporated into structures with composition and dimensions similar to natural bilayer membranes. The techniques reported here are applicable for coupling membrane receptors to a variety of transducing substrates used in biosensors.