title: Water-soluble acetylcholine receptor from Torpedo californica. Solubilization, purification and characterization.
abstract: The nicotinic acetylcholine receptor from electrogenic tissue of Torpedo californica was solubilized by tryptic digestion of membrane fragments obtained from autolysed tissue, without use of detergent. The water-soluble acetylcholine receptor was purified by affinity chromatography on a cobra-toxin-Sepharose resin. The purified receptor bound 4000-6000 pmol per mg protein of alpha-[125I]bungarotoxin, and toxin-binding was specifically inhibited by cholinergic ligands. Gel filtration revealed a single molecular species of Stokes radius 125 +/- 10 A and on sucrose gradient centrifugation one major peak was observed of 20-22 S. Polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate and beta-mercaptoethanol revealed two major polypeptides of mol. wt. 30 000 and 48 000.
authors.affiliation: Department of Neurobiology, Weizmann Institute of Science, Rehovot, Israel.
authors.forename: N
authors.initials: N
authors.lastname: Kalderon
authors.forename: I
authors.initials: I
authors.lastname: Silman
mesh_terms: D000818:Animals; D001703:Biophysics / Q000379:methods; D002038:Bungarotoxins / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D002499:Centrifugation, Density Gradient; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D003902:Detergents / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D020128:Inhibitory Concentration 50; D008623:Mercaptoethanol / Q000494:pharmacology; D010455:Peptides / Q000737:chemistry; D011950:Receptors, Cholinergic / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D012685:Sepharose / Q000494:pharmacology; D013395:Sucrose / Q000494:pharmacology; D014101:Torpedo; D014867:Water / Q000737:chemistry
chemical_list: D002038:Bungarotoxins; D003902:Detergents; D010455:Peptides; D011950:Receptors, Cholinergic; D014867:Water; D013395:Sucrose; D008623:Mercaptoethanol; D012685:Sepharose