通过用抗受体抗体标记证明乙酰胆碱受体的跨膜性质。
Demonstration of the transmembrane nature of the acetylcholine receptor by labeling with anti-receptor antibodies.
作者信息
Strader C B, Revel J P, Raftery M A
出版信息
J Cell Biol. 1979 Nov;83(2 Pt 1):499-510. doi: 10.1083/jcb.83.2.499.
Abstract
Antibodies raised in rabbits to Triton-solubilized, purified acetylcholine receptor from Torpedo californica were used to immunospecifically label intact T. californica electroplaque membrane vesicles attached to cover slips and oriented with the extracellular face of the synaptic membrane facing outward. Hemocyanin conjugated to Protein A was then used as a marker, making the antibody binding visible at the electron microscope level. Parallel labeling experiments were performed on vesicles attached to cover slips and sheared by sonication, leaving their cytoplasmic faces fully exposed to the labeling solution. While differences in antibody populations among different rabbits were observed, antigenic determinants of the receptor were present on both faces of the membrane, with those on the extracellular side more numerous than those on the cytoplasmic side, demonstrating the transmembrane nature of the receptor molecule.
摘要
用兔制备的针对来自加州电鳐经Triton溶解并纯化的乙酰胆碱受体的抗体，对附着在盖玻片上且突触膜细胞外表面朝外定向的完整加州电鳐电板膜囊泡进行免疫特异性标记。然后将与蛋白A偶联的血蓝蛋白用作标记物，使抗体结合在电子显微镜水平可见。对附着在盖玻片上并经超声处理剪切、使其细胞质面完全暴露于标记溶液的囊泡进行了平行标记实验。虽然观察到不同兔子之间抗体群体存在差异，但受体的抗原决定簇在膜的两面均有，细胞外侧的比细胞质侧的更多，这证明了受体分子的跨膜性质。
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