Purification of acetylcholine receptor-like protein from fetal calf thymus.
A cobrotoxin binding protein from the fetal calf thymus was isolated by affinity chromatography after solubilization with sodium cholate. The specific activity as a nicotinic acetylcholine receptor (AChR) was determined by assessing the binding to [3H]-alpha-bungarotoxin (BuTx), using the high-pressure liquid chromatography. An AChR-like protein was detected in the amount of 1.39-2.14 nmol per g protein. The first peak of 420k-protein from gel filtration of the eluate of affinity chromatography on a Sephacryl column showed one major polypeptide band with an Mr of 40k, by polyacrylamide gel electrophoresis in sodium dodecylsulfate, two major protein bands with pI 5.4-5.6 and 9.2 by isoelectric focusing, and reacted with sera from patients with myasthenia gravis.
D000818:Animals; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D007525:Isoelectric Focusing; D011506:Proteins / Q000302:isolation & purification; D011950:Receptors, Cholinergic / Q000302:isolation & purification*; D013950:Thymus Gland / Q000032:analysis*
D011506:Proteins; D011950:Receptors, Cholinergic