Bekele-Arcuri Z, Matos M F, Manganas L, Strassle B W, Monaghan M M, Rhodes K J, Trimmer J S
Department of Biochemistry and Cell Biology, State University of New York, Stony Brook 11794, USA.
Neuropharmacology. 1996;35(7):851-65. doi: 10.1016/0028-3908(96)00128-1.
Molecular characterization of mammalian voltage-sensitive K+ channel genes and their expression became possible with the cloning of the Shaker locus of Drosophila. However, analysis of the expression patterns and subunit composition of native K+ channel protein complexes requires immunological probes specific for the individual K+ channel gene products expressed in excitable tissue. Here, we describe the generation and characterization of monoclonal antibodies (mAbs) against eight distinct mammalian K+ channel polypeptides; the Kv1.1, Kv1.2, Kv1.4, Kv1.5 and Kv1.6 Shaker-related alpha-subunits, the Kv2.1 Shab-related alpha-subunit, and the Kv beta 1 and Kv beta 2 beta-subunits. We characterized the subtype-specificity of these mAbs against native K+ channels in mammalian brain and against recombinant K+ channels expressed in transfected mammalian cells. In addition, we used these mAbs to investigate the cellular and subcellular distribution of the corresponding polypeptides in rat cerebral cortex, as well as their expression levels across brain regions.
随着果蝇摇椅基因座的克隆，哺乳动物电压敏感性钾离子通道基因的分子特征及其表达成为可能。然而，分析天然钾离子通道蛋白复合物的表达模式和亚基组成需要针对可兴奋组织中表达的单个钾离子通道基因产物的免疫探针。在此，我们描述了针对八种不同哺乳动物钾离子通道多肽的单克隆抗体（mAb）的产生和特性；Kv1.1、Kv1.2、Kv1.4、Kv1.5和Kv1.6摇椅相关α亚基、Kv2.1沙布相关α亚基以及Kvβ1和Kvβ2β亚基。我们表征了这些单克隆抗体针对哺乳动物大脑中的天然钾离子通道以及在转染的哺乳动物细胞中表达的重组钾离子通道的亚型特异性。此外，我们使用这些单克隆抗体研究了大鼠大脑皮层中相应多肽的细胞和亚细胞分布，以及它们在不同脑区的表达水平。
