Reconstitution of nicotinic acetylcholine receptors into gel-protected lipid membranes

We report the functional reconstitution of nicotinic acetylcholine receptors into gel-protected bilayer lipid membranes using two different methods. In the first case, reconstitution was achieved by direct membrane formation from an emulsion of glycerol monooleate, hexane, and a membrane receptor extract. In the second case, incorporation was achieved via the fusion of vesicles from a preparation of membrane-bound receptors into preformed membranes after diffusion through the protective front gel layer. Measurement of the dc conductivity of the membranes in the presence of either acetylcholine or a-bungarotoxin was used to test for the functional activity of incorporated receptors.