Rosemblatt M S, Pérez G, Jaimovich E
Laboratorio de Inmunología Celular, INTA Universidad de Chile, Santiago.
Mol Cell Biochem. 1991 Aug 14;106(2):99-107. doi: 10.1007/BF00230175.
We have established several hybridoma lines that produce monoclonal antibodies against transverse tubule (t-tubule) proteins from frog skeletal muscle. The specificity of these antibodies was characterized by ELISA and Western immunoblotting with purified t-tubule, sarcoplasmic reticulum and partially purified sarcolemmal membranes. One of the monoclonal antibodies (2/34.4) recognizes a band of 109,000 Da in immunoblots. When purified frog t-tubule vesicles were preincubated with this antibody we observed an increase in the rate of the Mg(2+)-ATPase enzyme (up to six fold) which was dependent on antibody concentration. Immunocytological experiments done on cryostat sections of frog muscle indicate that the antigen recognized by this antibody is localized mainly at the level of the t-tubules (I band) and to a lesser extent at the sarcolemma. These results indicate that monoclonal antibody 2/34.4 recognizes the t-tubule Mg(2+)-ATPase and modulates its activity. This antibody should be useful as a probe on studies designed to study the physiological function of the enzyme.
我们已经建立了几个杂交瘤细胞系，它们能产生针对青蛙骨骼肌横小管（T小管）蛋白的单克隆抗体。通过酶联免疫吸附测定（ELISA）以及使用纯化的T小管、肌浆网和部分纯化的肌膜进行蛋白质免疫印迹分析，对这些抗体的特异性进行了表征。其中一种单克隆抗体（2/34.4）在免疫印迹中识别出一条109,000道尔顿的条带。当纯化的青蛙T小管囊泡与该抗体预孵育时，我们观察到镁离子-ATP酶的活性速率增加（高达六倍），且该增加依赖于抗体浓度。对青蛙肌肉冰冻切片进行的免疫细胞化学实验表明，该抗体识别的抗原主要定位于T小管（I带）水平，在肌膜上的定位程度较低。这些结果表明，单克隆抗体2/34.4识别T小管镁离子-ATP酶并调节其活性。该抗体可用作研究该酶生理功能的探针。
