Water-soluble acetylcholine receptor from Torpedo californica. Solubilization, purification and characterization.
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.
Kalderon
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N
Department of Neurobiology, Weizmann Institute of Science, Rehovot, Israel.
Silman
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I
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
D002038:Bungarotoxins; D003902:Detergents; D010455:Peptides; D011950:Receptors, Cholinergic; D014867:Water; D013395:Sucrose; D008623:Mercaptoethanol; D012685:Sepharose