胎牛胸腺中烟碱型乙酰胆碱受体样蛋白的分离与鉴定

Isolation and characterization of nicotinic acetylcholine receptor-like protein from fetal calf thymus.

作者信息

Kawanami S, Conti-Tronconi B, Racs J, Raftery M A

机构信息

Division of Biology and Chemical Engineering, California Institute of Technology, Pasadena 91125.

出版信息

J Neurol Sci. 1988 Nov;87(2-3):195-209. doi: 10.1016/0022-510x(88)90245-6.

DOI:10.1016/0022-510x(88)90245-6

PMID:3210032

Abstract

A nicotinic acetylcholine receptor-like protein (AChR-LP) was isolated from fetal calf thymus by affinity chromatography using cobrotoxin-Sepharose after alkaline extraction and solubilization with Triton X-100. The AChR-LP had a specificity of 1.61 +/- 1.12 nmol of alpha-bungarotoxin binding sites per mg of protein. The isoelectric point, sedimentation coefficient and amino acid composition of the purified AChR-LP were very similar to those of muscle and electric organ AChRs. Upon SDS-polyacrylamide gel electrophoresis purified thymus AChR-LP preparations contained up to 6 polypeptide bands of molecular weights of 40,000, 43,000, 51,000, 56,000, 58,000, and 66,000, respectively. The peptides of 40,000, 51,000, 56,000, and 66,000 dalton cross-reacted with the four subunits of Torpedo californica and fetal calf muscle AChR.

摘要

通过碱性提取并用Triton X-100溶解后，利用眼镜蛇毒素-琼脂糖亲和层析从胎牛胸腺中分离出一种烟碱型乙酰胆碱受体样蛋白（AChR-LP）。AChR-LP的特异性为每毫克蛋白质1.61±1.12纳摩尔的α-银环蛇毒素结合位点。纯化后的AChR-LP的等电点、沉降系数和氨基酸组成与肌肉和电器官的AChR非常相似。在SDS-聚丙烯酰胺凝胶电泳中，纯化的胸腺AChR-LP制剂分别含有多达6条分子量为40,000、43,000、51,000、56,000、58,000和66,000的多肽条带。分子量为40,000、51,000、56,000和66,000道尔顿的肽与加州电鳐和胎牛肌肉AChR的四个亚基发生交叉反应。