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首页> 外文期刊>The European Journal of Neuroscience >Evidence for oligomerization between GABAB receptors and GIRK channels containing the GIRK1 and GIRK3 subunits.
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Evidence for oligomerization between GABAB receptors and GIRK channels containing the GIRK1 and GIRK3 subunits.

机译：GABAB受体与包含GIRK1和GIRK3亚基的GIRK通道之间低聚的证据。

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摘要

The stimulation of inhibitory neurotransmitter receptors, such as gamma-aminobutyric acid type B (GABA(B) ) receptors, activates G protein-gated inwardly-rectifying K(+) (GIRK) channels, which influence membrane excitability. There is now evidence suggesting that G protein-coupled receptors and G protein-gated inwardly-rectifying K(+) [GIRK/family 3 of inwardly-rectifying K(+) (Kir3)] channels do not diffuse freely within the plasma membrane, but instead there are direct protein-protein interactions between them. Here, we used bioluminescence resonance energy transfer, co-immunoprecipitation, confocal and electron microscopy techniques to investigate the oligomerization of GABA(B) receptors with GIRK channels containing the GIRK3 subunit, whose contribution to functional channels is still unresolved. Co-expression of GABA(B) receptors and GIRK channels in human embryonic kidney-293 cells in combination with co-immunoprecipitation experiments established that the metabotropic receptor forms stable complexes with GIRK channels. Using bioluminescence resonance energy transfer, we have shown that, in living cells under physiological conditions, GABA(B) receptors interact directly with GIRK1/GIRK3 heterotetramers. In addition, we have provided evidence that the receptor-effector complexes are also found in vivo and identified that the cerebellar granule cells are one neuron population where the interaction probably takes place. Altogether, our data show that signalling complexes containing GABA(B) receptors and GIRK channels are formed shortly after biosynthesis, probably in the endoplasmic reticulum and/or endoplasmic reticulum/Golgi apparatus complex, suggesting that this might be a general feature of receptor-effector ion channel signal transduction and supporting a channel-forming role for the GIRK3 subunit.
机译：抑制性神经递质受体的刺激，例如B型γ-氨基丁酸（GABA（B））受体，激活G蛋白门控的内向整流K（+）（GIRK）通道，从而影响膜的兴奋性。现在有证据表明，G蛋白偶联受体和G蛋白门控的向内整流K（+）[GIRK /向内整流K（+）（Kir3）的家族3]不能在质膜内自由扩散，但是相反，它们之间存在直接的蛋白质-蛋白质相互作用。在这里，我们使用生物发光共振能量转移，免疫共沉淀，共聚焦和电子显微镜技术研究GABA（B）受体与含有GIRK3亚基的GIRK通道的寡聚化，其对功能通道的贡献仍未解决。 GABA（B）受体和GIRK通道在人胚肾293细胞中的共表达与免疫共沉淀实验相结合，建立了促代谢受体与GIRK通道形成稳定的复合物。使用生物发光共振能量转移，我们已经表明，在生理条件下的活细胞中，GABA（B）受体与GIRK1 / GIRK3异四聚体直接相互作用。此外，我们已经提供了证据，表明体内还发现了受体效应物复合物，并确定了小脑颗粒细胞是可能发生相互作用的一种神经元群体。总的来说，我们的数据表明，包含GABA（B）受体和GIRK通道的信号复合物在生物合成后不久就形成了，可能是在内质网和/或内质网/高尔基体复合物中形成的，这表明这可能是受体效应子的一般特征。离子通道信号转导并支持GIRK3亚基的通道形成作用。

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