Babel D, Walter G, Götz H, Thinnes F P, Jürgens L, König U, Hilschmann N
Max-Planck-Institute für experimentelle Medizin, Abteilung Immunochemie, Göttingen.
Biol Chem Hoppe Seyler. 1991 Dec;372(12):1027-34. doi: 10.1515/bchm3.1991.372.2.1027.
We report on the production and characterization of eight monoclonal mouse antibodies against the complete human VDAC "Porin 31HL". The antigen used was purified from a total membrane preparation of the transformed human B-lymphocyte cell line H2LCL. In Western blots all eight mAbs react with a single 31-kDa band in solubilized H2LCL membrane preparations thus demonstrating their specificity for the human VDAC "Porin 31HL". Concerning the epitope specificity we show that all eight mAbs equally react with the N-terminal part of human porin. Moreover, we demonstrate the expression of VDAC in the sarcolemma by indirect immunoenzyme labelling of cryosections of human skeletal muscle applying four of our mAbs. These data support our recent observations on the expression of porin channels in the plasmalemma of different normal and transformed human cell lines. VDAC in the plasmalemma is discussed as the molecular basis of the Blatz and Magleby channel.
我们报告了针对完整的人电压依赖性阴离子通道“孔蛋白31HL”的八种单克隆小鼠抗体的制备及特性鉴定。所用抗原是从转化的人B淋巴细胞系H2LCL的全细胞膜制剂中纯化得到的。在蛋白质免疫印迹中，所有八种单克隆抗体都与溶解的H2LCL细胞膜制剂中的一条31 kDa的条带发生反应，从而证明了它们对人电压依赖性阴离子通道“孔蛋白31HL”的特异性。关于表位特异性，我们表明所有八种单克隆抗体与人孔蛋白的N端部分均有同等反应。此外，我们通过应用我们的四种单克隆抗体对人骨骼肌冷冻切片进行间接免疫酶标记，证明了电压依赖性阴离子通道在肌膜中的表达。这些数据支持了我们最近关于孔蛋白通道在不同正常和转化的人细胞系质膜中表达的观察结果。质膜中的电压依赖性阴离子通道被认为是布拉茨和马格利比通道的分子基础。
