通过共价交联对大鼠肺膜胰高血糖素样肽-I(7-36)酰胺受体进行表征

Characterization of glucagon-like peptide-I(7-36)amide receptors of rat lung membranes by covalent cross-linking.

作者信息

Richter G, Göke R, Göke B, Schmidt H, Arnold R

机构信息

Department of Internal Medicine, Philipps-University of Marburg, Germany.

出版信息

FEBS Lett. 1991 Mar 25;280(2):247-50. doi: 10.1016/0014-5793(91)80303-k.

DOI:10.1016/0014-5793(91)80303-k

PMID:1849486

Abstract

125I-labelled GLP-I(7-36)amide was cross-liked to a specific binding protein in rat lung membranes using disuccinimidyl suberate. A single radio-labelled band at Mr 66,000 was identified by SDS-PAGE after solubilization of the ligand-binding protein complex which is consistent with the presence of a single class of binding sites on rat lung membranes. The band was undetectable when 1 mumol/l GLP-I(7-36)amide was included in the binding assay. No change in the mobility of the band was observed under reducing conditions suggesting that the binding protein in the receptor is not part of a larger disulphide-linked protein. The intensity of the radiolabelled protein band was reduced when the incubation with 125I-labelled GLP-I(7-36)amide was carried out in the presence of guanine nucleotides suggesting that the GLP-I(7-36)amide receptor is coupled to the adenylate cyclase system.

摘要

使用辛二酸二琥珀酰亚胺酯将125I标记的胰高血糖素样肽-1（7-36）酰胺与大鼠肺膜中的一种特异性结合蛋白进行交联。在溶解配体-结合蛋白复合物后，通过SDS-PAGE鉴定出一条分子量为66,000的单一放射性标记条带，这与大鼠肺膜上存在单一类别的结合位点一致。当在结合试验中加入1 μmol/L的胰高血糖素样肽-1（7-36）酰胺时，该条带无法检测到。在还原条件下未观察到条带迁移率的变化，这表明受体中的结合蛋白不是更大的二硫键连接蛋白的一部分。当在鸟嘌呤核苷酸存在的情况下与125I标记的胰高血糖素样肽-1（7-36）酰胺一起孵育时，放射性标记蛋白条带的强度降低，这表明胰高血糖素样肽-1（7-36）酰胺受体与腺苷酸环化酶系统偶联。