Generation and characterization of subtype-specific monoclonal antibodies to K+ channel alpha- and beta-subunit polypeptides
Generation and characterization of subtype-specific monoclonal antibodies to K+ channel alpha- and beta-subunit polypeptides
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DOI：
10.1016/0028-3908(96)00128-1
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发表时间：
1996-07-01
影响因子：
4.7
通讯作者：
Trimmer, JS
中科院分区：
文献类型：
--
作者：
BekeleArcuri, Z;Matos, MF;Trimmer, JS
随着果蝇Shaker基因座的克隆，哺乳动物电压敏感K+通道基因的分子特征及其表达成为可能。然而，分析天然K+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K+通道基因产物的免疫探针。在这里，我们描述了针对八种不同的哺乳动物K+通道多肽的单克隆抗体（mab）的产生和表征；Kv1.1、Kv1.2、Kv1.4、Kv1.5和Kv1.6与shaker相关的α -亚基，Kv2.1与shab相关的α -亚基，Kv β 1和Kv β 2 β亚基。我们鉴定了这些单克隆抗体对哺乳动物大脑中天然K+通道和转染的哺乳动物细胞中表达的重组K+通道的亚型特异性。此外，我们利用这些单克隆抗体研究了相应多肽在大鼠大脑皮层的细胞和亚细胞分布，以及它们在大脑区域的表达水平。版权所有爱思唯尔科学有限公司
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. Copyright (C) 1996 Elsevier Science Ltd