The alpha-bungarotoxin-binding nicotinic acetylcholine receptor from rat brain contains only the alpha 7 subunit
The alpha-bungarotoxin-binding nicotinic acetylcholine receptor from rat brain contains only the alpha 7 subunit
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DOI：
10.1074/jbc.272.38.24024
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发表时间：
1997-09-19
影响因子：
4.8
通讯作者：
Patrick, JW
中科院分区：
文献类型：
--
作者：
Chen, DN;Patrick, JW
当在爪蟾卵母细胞中表达时，大鼠α - 7亚基形成同源低聚烟碱乙酰胆碱受体，该受体被α -班加罗毒素阻断。由于在卵母细胞中表达的α - 7受体的药理学和生理学特性与从神经元制剂中记录的α -班加罗毒素敏感烟碱电流相似，并且α - 7 mRNA和α -班加罗毒素结合位点在大鼠脑中的分布模式非常相似，因此α - 7被认为是哺乳动物大脑中α -班加罗毒素结合烟碱受体的主要成分。然而，尽管从大鼠脑或PC12细胞中纯化的α -班加罗毒素结合复合物中发现了α - 7，但其他蛋白质会与它共化。因此，α - 7是否在哺乳动物大脑中形成α -班加罗毒素结合的同质寡聚烟碱受体的问题仍然存在。我们已经开发并鉴定了亲和纯化的多克隆抗体，并将这些抗体用于从大鼠脑中纯化的α -班加罗毒素结合蛋白的Western blot分析。我们在此报告，我们的实验数据支持目前的工作假设，即α -班加罗毒素结合的烟碱受体是大鼠大脑中α 7亚基的同质寡聚物。
When expressed in Xenopus oocytes, the rat alpha 7 subunit forms homo-oligomeric nicotinic acetylcholine receptors, which are blocked by alpha-bungarotoxin. Since the pharmacological and physiological properties of the alpha 7 receptor expressed in oocytes are similar to those of the alpha-bungarotoxin-sensitive nicotinic currents recorded from neuronal preparations and the distribution patterns of alpha 7 mRNA and alpha-bungarotoxin-binding sites in the rat brain are very similar, alpha 7 is thought to be the main component of the alpha-bungarotoxin-binding nicotinic receptor in the mammalian brain. However, while alpha 7 is found in purified alpha-bungarotoxin-binding complexes from rat brain or PC12 cells, other proteins copurify with it. Therefore, the question whether alpha 7 forms a homo-oligomeric alpha-bungarotoxin-binding nicotinic receptor in the mammalian brain remains. We have developed and characterized affinity-purified polyclonal antibodies and used these antibodies in Western blot analyses of alpha-bungarotoxin-binding proteins purified from rat brains. We report here that our experimental data support the current working hypothesis that the alpha-bungarotoxin-binding nicotinic receptor is a homo-oligomer of alpha 7 subunits in the rat brain.