...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Imaging of receptor trafficking by using α-bungarotoxin-binding-site-tagged receptors
【24h】

Imaging of receptor trafficking by using α-bungarotoxin-binding-site-tagged receptors

机译：通过使用α-真菌毒素结合位点标记的受体对受体运输进行成像

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

α-Amino-3-hydroxy-5-methyl-4-isoxazole-propionate (AMPA) receptors mediate excitatory synaptic transmission and are dynamically regulated during synaptic plasticity in the CNS. The membrane trafficking of AMPA receptors to synapses is critical for the regulation of the efficacy of excitatory synaptic transmission. Direct imaging of AMPA receptors in various cell compartments is important to dissecting the regulation of distinct steps in receptor membrane trafficking. In this study, we have developed an approach for the imaging of receptor trafficking with subunits tagged with a 13-aa α-bungarotoxin (BTX)-binding site (BBS). The small polypeptide neurotoxin BTX has been used for decades to study the nicotinic acetylcholine receptor. Similar high-affinity ligands are rarely available for most receptors. Engineering the BBS tag into receptor subunits allowed the high-affinity binding of fluorescent, radioactive, and biotinylated BTX to the tagged receptor subunits. By using this approach, the total receptor expression, surface expression, internalization, and insertion of receptors into the plasma membrane could be visualized and quantified in fixed or live cells including cultured neurons. The BBS tag is a flexible approach for labeling membrane proteins and studying their dynamic trafficking.
机译：α-Amino-3-hydroxy-5-methyl-4-isoxazole-propionate（AMPA）受体介导兴奋性突触传递，并在CNS的突触可塑性过程中受到动态调节。 AMPA受体的膜运输到突触对于调节兴奋性突触传递的功效至关重要。在各个细胞区室中对AMPA受体进行直接成像对于剖析受体膜运输中不同步骤的调控非常重要。在这项研究中，我们已经开发出一种带有13-aaα-真菌毒素（BTX）结合位点（BBS）标记的亚基的受体运输成像方法。小型多肽神经毒素BTX已用于研究烟碱型乙酰胆碱受体数十年。类似的高亲和力配体很少可用于大多数受体。将BBS标签改造成受体亚基可以使荧光，放射性和生物素化的BTX高亲和力结合到标记的受体亚基上。通过使用这种方法，可以在包括培养的神经元在内的固定或活细胞中可视化并量化受体的总表达，表面表达，内在化以及将受体插入质膜中。 BBS标签是一种用于标记膜蛋白并研究其动态运输的灵活方法。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号