Reconstitution of the nicotinic acetylcholine receptor using a lipid substitution technique.
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.
Jones
O T
OT
Department of Biochemistry and Biophysics, University of California, Davis 95616.
Eubanks
J H
JH
Earnest
J P
JP
McNamee
M G
MG
D000818:Animals; D002499:Centrifugation, Density Gradient; D003902:Detergents; D066298:In Vitro Techniques; D007473:Ion Channels / Q000502:physiology; D046911:Macromolecular Substances; D008563:Membrane Lipids / Q000378:metabolism*; D010713:Phosphatidylcholines; D011978:Receptors, Nicotinic / Q000502:physiology*; D013329:Structure-Activity Relationship; D014101:Torpedo
D003902:Detergents; D007473:Ion Channels; D046911:Macromolecular Substances; D008563:Membrane Lipids; D010713:Phosphatidylcholines; D011978:Receptors, Nicotinic