Interaction of the 43K protein with components of Torpedo postsynaptic membranes.
Interactions of the major Mr 43 000 peripheral membrane protein (43K protein) with components of Torpedo postsynaptic membranes have been examined. Treatment of membranes with copper o-phenanthroline promotes the polymerization of 43K protein to dimers and higher oligomers. These high molecular weight forms of 43K protein can be converted to monomers by reduction with dithiothreitol and do not contain any of the other major proteins found in these membranes, including the subunits of the acetylcholine receptor, as shown by immunoblotting with monoclonal antibodies. To study directly its interactions with the membrane, the 43K protein was radioiodinated and purified by immunoaffinity chromatography. Purified 43K protein binds tightly to pure liposomes of various compositions in a manner that is not inhibited by KCl concentrations up to 0.75 M. The binding can be reversed by adjusting the pH of the reaction to 11, the same treatment that removes 43K protein from postsynaptic membranes. Unlabeled 43K protein solubilized from Torpedo membranes with cholate can be reconstituted with exogenously added lipids in the absence of the receptor. The results suggest that 43K protein molecules are amphipathic and that they may interact with each other and with the lipid bilayer. These interactions cannot explain the coextensive distribution of 43K proteins with acetylcholine receptors in situ. However, they could account for the association of the 43K protein with the postsynaptic membrane and may contribute to the maintenance of the structure of the cytoplasmic specialization of which this protein is a major component.
D000818:Animals; D003432:Cross-Linking Reagents; D008081:Liposomes; D046911:Macromolecular Substances; D008565:Membrane Proteins / Q000378:metabolism*; D008970:Molecular Weight; D009469:Neuromuscular Junction / Q000648:ultrastructure; D011485:Protein Binding; D011950:Receptors, Cholinergic / Q000378:metabolism*; D013570:Synaptic Membranes / Q000378:metabolism*; D014101:Torpedo
D003432:Cross-Linking Reagents; D008081:Liposomes; D046911:Macromolecular Substances; D008565:Membrane Proteins; D011950:Receptors, Cholinergic