Pfeiffer F, Simler R, Grenningloh G, Betz H
Proc Natl Acad Sci U S A. 1984 Nov;81(22):7224-7. doi: 10.1073/pnas.81.22.7224.
The glycine receptor of rat spinal cord is an oligomeric membrane glycoprotein of molecular mass 250,000 daltons that contains three polypeptides of 48,000, 58,000, and 93,000 daltons. Monoclonal antibodies (mAbs) were prepared against the affinity-purified glycine receptor protein by using 125I-labeled receptor preparations for the detection of positive hybrids. From nine monoclonal antibodies obtained, six recognized denatured receptor polypeptides blotted to nitrocellulose paper. Two of these antibodies bound to more than one glycine receptor polypeptide: mAb GlyR 4a stained the 48,000- and 58,000-dalton polypeptides, and mAb GlyR 7a stained the 48,000- and 93,000-dalton polypeptides. Common antigenic determinants thus are shared by the different subunits of the glycine receptor. Complementary results were obtained by peptide mapping of 125I-labeled glycine receptor polypeptides with various proteases. A set of peptide fragments of the same apparent molecular mass was produced from the different glycine receptor polypeptides by using V8 protease, chymotrypsin, and elastase. These data suggest that the subunits of the glycine receptor have significant homology within their primary structure and may have evolved from a common ancestor receptor polypeptide.
大鼠脊髓中的甘氨酸受体是一种分子量为250,000道尔顿的寡聚膜糖蛋白，它包含三条分子量分别为48,000、58,000和93,000道尔顿的多肽。通过使用125I标记的受体制剂来检测阳性杂交体，制备了针对亲和纯化的甘氨酸受体蛋白的单克隆抗体（mAb）。从获得的九种单克隆抗体中，有六种识别印迹在硝酸纤维素纸上的变性受体多肽。其中两种抗体与不止一种甘氨酸受体多肽结合：单克隆抗体GlyR 4a对48,000道尔顿和58,000道尔顿的多肽染色，单克隆抗体GlyR 7a对48,000道尔顿和93,000道尔顿的多肽染色。因此，甘氨酸受体的不同亚基共享共同的抗原决定簇。通过用各种蛋白酶对125I标记的甘氨酸受体多肽进行肽图谱分析，获得了互补的结果。使用V8蛋白酶、胰凝乳蛋白酶和弹性蛋白酶，从不同的甘氨酸受体多肽中产生了一组表观分子量相同的肽片段。这些数据表明，甘氨酸受体的亚基在其一级结构内具有显著的同源性，并且可能从一个共同的祖先受体多肽进化而来。
