从胎牛胸腺中纯化类乙酰胆碱受体蛋白。

Purification of acetylcholine receptor-like protein from fetal calf thymus.

作者信息

Kawanami S, Kakuno T, Horio T

出版信息

Jpn J Psychiatry Neurol. 1987 Mar;41(1):97-104. doi: 10.1111/j.1440-1819.1987.tb00396.x.

DOI:10.1111/j.1440-1819.1987.tb00396.x

Abstract

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.

摘要

用胆酸钠溶解后，通过亲和色谱法从胎牛胸腺中分离出一种眼镜蛇毒素结合蛋白。通过高压液相色谱法评估其与[3H]-α-银环蛇毒素（BuTx）的结合，从而确定其作为烟碱型乙酰胆碱受体（AChR）的比活性。检测到一种类似AChR的蛋白质，每克蛋白质含量为1.39 - 2.14 nmol。在Sephacryl柱上对亲和色谱洗脱液进行凝胶过滤，得到的420k-蛋白的第一个峰，经十二烷基硫酸钠聚丙烯酰胺凝胶电泳显示有一条主要的多肽带，Mr为40k，等电聚焦显示有两条主要蛋白带，pI分别为5.4 - 5.6和9.2，并且与重症肌无力患者的血清发生反应。