Gorvel J P, Liabeuf A, Massey D, Liot D, Goridis C, Maroux S
Cell Tissue Res. 1983;234(3):619-32. doi: 10.1007/BF00218655.
The antigen detected by the rat anti-mouse monoclonal antibody (m Ab), anti-BSP-3, has been initially described as a brain cell-surface protein. Evidence is presented that this m Ab recognizes mouse (Na+ + K+)-ATPase (ATP phosphohydrolase, E.C.3.6.1.3). The antigen, purified from mouse brain by means of affinity chromatography, migrated in SDS-polyacrylamide gels in the form of two polypeptide chains of 100 000 and 48 000 molecular weight, which could be shown to react with subunit-specific polyclonal antisera against ATPase in immunoblotting experiments. Purified BSP-3 antigen was bound to the specific (Na+ + K+)-ATPase inhibitor ouabain. Finally, the anti-BSP-3 m Ab was capable of immunoprecipitating the ATPase activity of a microsomal fraction from mouse kidney. The m Ab was used to study the localization of (Na+ + K+)-ATPase in different organs of the mouse. It stained the basolateral plasma membranes of polarized cells in immunofluorescence experiments, while the entire cell surface of unpolarized cells was labeled. Interestingly, several cell types did not react with the m Ab, indicating a possible heterogeneity of ATPases. Such a m Ab could prove to be a useful tool for studying localization, structure and function of (Na+ + K+)-ATPase.
大鼠抗小鼠单克隆抗体（mAb）抗BSP-3所检测到的抗原最初被描述为一种脑细胞表面蛋白。本文提供的证据表明，这种mAb识别小鼠（Na⁺+K⁺）-ATP酶（ATP磷酸水解酶，E.C.3.6.1.3）。通过亲和层析从小鼠脑中纯化得到的该抗原，在SDS-聚丙烯酰胺凝胶中以分子量为100000和48000的两条多肽链形式迁移，在免疫印迹实验中可证明其能与针对ATP酶的亚基特异性多克隆抗血清发生反应。纯化的BSP-3抗原与特异性（Na⁺+K⁺）-ATP酶抑制剂哇巴因结合。最后，抗BSP-3 mAb能够免疫沉淀小鼠肾脏微粒体部分的ATP酶活性。该mAb被用于研究小鼠不同器官中（Na⁺+K⁺）-ATP酶的定位。在免疫荧光实验中，它可对极化细胞的基底外侧质膜进行染色，而未极化细胞的整个细胞表面都被标记。有趣的是，几种细胞类型不与该mAb反应，这表明ATP酶可能存在异质性。这样一种mAb可能被证明是研究（Na⁺+K⁺）-ATP酶定位、结构和功能的有用工具。
