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The translational diffusion of bovine rhodopsin, the Ca2+-activated adenosinetriphosphatase of rabbit muscle sarcoplasmic reticulum, and the acetylcholine receptor monomer of Torpedo marmorata has been examined at a high dilution (molar ratios of lipid/protein greater than or equal to 3000/1) in liquid-crystalline phase phospholipid bilayer membranes by(More)
Membranes of cellular organelles and plasma membranes of some type of cells are not accessible to the high-resolution recordings that the conventional patch-clamp technique allows. However, when these purified membranes are dehydrated together with small lipid vesicles and hydrated again, cell-size vesicles (5-100 micron diameter) are obtained, on which(More)
Torpedo acetylcholine receptor was reconstituted into liposomes of pure synthetic lipids in order to study the influence of the lipid environment on affinity state transitions and the ion translocation function of the receptor. A critical concentration of 30 to 40% of cholesteryl hemisuccinate was necessary in liposomes made of cholesteryl hemisuccinate and(More)
The muscular content of the mRNAs encoding the five subunits of the nicotonic acetylcholine receptor was measured during postnatal development in the rat. Subunit specific mRNAs show differential regulation. The levels of the alpha-, gamma- and delta-subunit specific mRNAs decrease steadily after birth, while the beta- and epsilon-subunit mRNAs increase(More)
The isolation and characterization of five clones carrying sequences of the alpha-, beta-, gamma-, delta- and epsilon-subunit precursors of the rat muscle acetylcholine receptor (AChR) are described. The deduced amino acid sequences indicate that these polypeptides contain 457-519 amino acids and reveal the structural characteristics common to subunits of(More)
The effects of lipids on the maintenance of characteristic functional properties of the acetylcholine receptor during the course of reconstitution into lipid vesicles were studied by following the kinetics of agonist-induced state transitions. The requirements for successful preservation of these properties could be dissected into two components: (a)(More)
Single channel recording of reconstituted ion channels is possible by patch clamp measurements of giant liposomes formed by dehydration-rehydration of lipid films. This "hydration technique" consists of carefully controlled dehydration of a suspension of small vesicles followed by rehydration of the residue resulting in formation of large liposomes. Patch(More)
NaIO(4) oxidation, exo- and endo-glycosidase treatments and combinations thereof have been applied to acetylcholine receptor from Torpedo marmorata in its membrane-bound and detergent-solubilised forms. The effects of these chemical and enzymatic treatments are made apparent in the electrophoretic properties of the four receptor subunits (?, ?, ? and ?) and(More)