Interaction of the Glycine Receptor Alpha 1 Binding Site with Partial Agonists
Interaction of the Glycine Receptor Alpha 1 Binding Site with Partial Agonists
复制标题
甘氨酸受体 Alpha 1 结合位点与部分激动剂的相互作用
DOI：
10.1016/j.bpj.2013.11.3047
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发表时间：
2014
影响因子：
3.4
通讯作者：
Greiner T
中科院分区：
文献类型：
--
作者：
Greiner T
甘氨酸受体是五聚体配体门控离子通道超家族的成员。在脊椎动物中，它介导快速抑制传递，特别是在脑干和脊髓中。在人类中已经克隆了几种α亚基，如α1、α2和α3，它们可能形成同源和异源通道。每个亚基包括一个跨膜结构域，形成通道孔，一个大的细胞外结构域，包含结合位点和细胞内结构域（Lynch，Neuropharmacology，56，303，2009年）。我们的目标是深入了解结构-甘氨酸受体的功能关系，通过表征它们的激活机制和测量激动剂结合和通道在其激活时经历的构象变化，激活。为此，我们使用膜片钳技术在细胞贴附配置低噪声单通道记录。其次是一个密集的分析，包括时间过程拟合理想化和测试假设的激活机制直接拟合记录的数据。另一种方法是使用激动剂浓度跳跃应用于外向配置补丁，以评估非平衡激活的通道。
The glycine receptor is a member of the pentameric ligand-gated ion channel superfamily. In vertebrates, it mediates fast inhibitory transmission, particularly in the brainstem and spinal cord. Several alpha subunits have been cloned in man, eg α1, α2 and α3, and are likely to form homomeric and heteromeric channels. Each subunit consists of a transmembrane domain, forming the channel pore, a large extracellular domain that contains the binding site and an intracellular domain (Lynch, Neuropharmacology, 56, 303, 2009).Our aim is to gain insight into the structure-function relation of the glycine receptor by characterising their activation mechanism and measuring agonist binding and the conformational changes that the channel undergoes as it activates. For this we use the patch-clamp technique in cell-attached configuration for low noise single-channel recordings. This is followed by an intensive analysis that includes time course fitting idealization and testing of postulated activation mechanisms by direct fitting to the recorded data. A further approach is the use of agonist concentration jumps applied to outside-out configuration patches to assess non equilibrium activation of the channel.