Site-specific contacts enable distinct modes of TRPV1 regulation by the potassium channel Kvβ1 subunit.
Site-specific contacts enable distinct modes of TRPV1 regulation by the potassium channel Kvβ1 subunit.
复制标题
位点特异性接触使钾通道 Kvβ1 亚基能够实现不同的 TRPV1 调节模式。
DOI：
10.1074/jbc.ra120.015605
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发表时间：
2020-12-11
期刊：
影响因子：
--
通讯作者：
Yao J
中科院分区：
文献类型：
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作者：
Wang Y;Mo X;Ping C;Huang Q;Zhang H;Xie C;Zhong B;Li D;Yao J
瞬时受体电位香草酸亚型1（TRPV1）通道是一种多模式受体，负责痛觉、热觉和机械感觉。然而，哪些生物分子伴侣与TRPV1发生特异性相互作用仍有待阐明。在此，我们采用小鼠背根神经节基因的cDNA文库筛选结合膜片钳电生理学方法，鉴定出与TRPV1通道发生物理相互作用并调节其功能的电压门控钾通道辅助亚基Kvβ1。利用内源性背根神经节神经元以及HEK 293T细胞中的重组表达模型，在原位验证了这种相互作用。Kvβ1的存在增强了TRPV1通道在质膜上的表达稳定性以及痛觉电流密度。令人惊讶的是，Kvβ1的相互作用还改变了TRPV1热激活的温度阈值。通过位点特异性定位，我们进一步揭示Kvβ1分别与TRPV1的膜远端结构域和膜近端结构域相互作用，以调节其膜表达和温度激活阈值。因此，我们的数据表明Kvβ1是TRPV1信号复合体中的关键元件，并以位点特异性方式发挥双重调节作用。
Transient receptor potential vanilloid 1 (TRPV1) channel is a multimodal receptor that is responsible for nociceptive, thermal, and mechanical sensations. However, which biomolecular partners specifically interact with TRPV1 remains to be elucidated. Here, we used cDNA library screening of genes from mouse dorsal root ganglia combined with patch-clamp electrophysiology to identify the voltage-gated potassium channel auxiliary subunit Kvβ1 physically interacting with TRPV1 channel and regulating its function. The interaction was validated in situ using endogenous dorsal root ganglia neurons, as well as a recombinant expression model in HEK 293T cells. The presence of Kvβ1 enhanced the expression stability of TRPV1 channels on the plasma membrane and the nociceptive current density. Surprisingly, Kvβ1 interaction also shifted the temperature threshold for TRPV1 thermal activation. Using site-specific mapping, we further revealed that Kvβ1 interacted with the membrane-distal domain and membrane-proximal domain of TRPV1 to regulate its membrane expression and temperature-activation threshold, respectively. Our data therefore suggest that Kvβ1 is a key element in the TRPV1 signaling complex and exerts dual regulatory effects in a site-specific manner.