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Reconstitution of the nicotinic acetylcholine receptor using a lipid substitution technique
O T Jones1, J H Eubanks, J P Earnest
1Department of Biochemistry and Biophysics, University of California, Davis 95616.
Abstract:
The nicotinic acetylcholine receptor was purified by affinity chromatography in the presence of dioleoylphosphatidylcholine (DOPC). A method for replacing the DOPC with other lipids was developed by using detergent solubilization with a large excess of the new lipid followed by sucrose density gradient centrifugation in detergent-free buffers to separate receptor-lipid complexes from excess lipid and detergent. Homogenous complexes of defined lipid composition could be easily prepared and the efficiency of substitution was independent of lipid type. However, the functional properties of the resulting lipid complexes depended on the lipid composition.