Regulation of PKD2 channel function by TACAN
Regulation of PKD2 channel function by TACAN
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TACAN 对 PKD2 通道功能的调节
DOI：
10.1113/jp283895
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发表时间：
2022-12-13
影响因子：
5.5
通讯作者：
Chen，Xing-Zhen
中科院分区：
文献类型：
--
作者：
Liu，Xiong;Zhang，Rui;Chen，Xing-Zhen
摘要常染色体显性多囊肾病是由膜受体PKD 1或阳离子通道PKD 2突变引起的。TACAN（也称为TMEM 120 A），最近被报道为神经元中用于机械感知和疼痛感知的离子通道，也分布在各种非神经元组织中，如肾脏，心脏和肠，表明其参与其他功能。在这项研究中，我们发现，在天然肾细胞系中，塔康与PKD 2形成复合物。在异种卵母细胞中使用双电极电压钳，我们发现TACAN抑制PKD 2功能获得突变体F604 P的通道活性。含有第一个和最后一个跨膜结构域的TACAN片段分别与PKD 2 C-和N-末端片段相互作用。TACAN N末端作为封闭肽，TACAN通过PKD 2与TACAN的结合抑制PKD 2的功能。通过哺乳动物细胞的膜片钳技术，我们发现塔康抑制PKD 2和突变体F604 P的单通道电导和开放概率。PKD 2与塔康共表达，但单独PKD 2不表达，表现出压力敏感性。此外，我们发现塔康使斑马鱼幼鱼中PKD 2依赖的尾弯曲和前肾囊肿变小。综上所述，本研究揭示了塔康作为PKD 2抑制剂，并介导PKD 2-塔康通道复合物的机械敏感性。关键点塔康在体外和体内抑制PKD 2的功能。塔康N末端含S1的片段T160 X与PKD 2 C末端片段N580-L700相互作用，其C末端含S6的片段L296-D343与PKD 2 N末端A594 X相互作用。TACAN通过物理相互作用抑制PKD 2通道的功能。PKD 2与TACAN的复合物，而不是单独的PKD 2，赋予机械敏感性。
AbstractAutosomal dominant polycystic kidney disease is caused by mutations in the membrane receptor PKD1 or the cation channel PKD2. TACAN (also termed TMEM120A), recently reported as an ion channel in neurons for mechanosensing and pain sensing, is also distributed in diverse non‐neuronal tissues, such as kidney, heart and intestine, suggesting its involvement in other functions. In this study, we found that TACAN is in a complex with PKD2 in native renal cell lines. Using the two‐electrode voltage clamp inXenopusoocytes, we found that TACAN inhibits the channel activity of PKD2 gain‐of‐function mutant F604P. TACAN fragments containing the first and last transmembrane domains interacted with the PKD2 C‐ and N‐terminal fragments, respectively. The TACAN N‐terminus acted as a blocking peptide, and TACAN inhibited the function of PKD2 by the binding of PKD2 with TACAN. By patch clamping in mammalian cells, we found that TACAN inhibits both the single‐channel conductance and the open probability of PKD2 and mutant F604P. PKD2 co‐expressed with TACAN, but not PKD2 alone, exhibited pressure sensitivity. Furthermore, we found that TACAN aggravates PKD2‐dependent tail curvature and pronephric cysts in larval zebrafish. In summary, this study revealed that TACAN acts as a PKD2 inhibitor and mediates mechanosensitivity of the PKD2–TACAN channel complex.Key pointsTACAN inhibits the function of PKD2in vitroandin vivo.TACAN N‐terminal S1‐containing fragment T160X interacts with the PKD2 C‐terminal fragment N580–L700, and its C‐terminal S6‐containing fragment L296–D343 interacts with the PKD2 N‐terminal A594X.TACAN inhibits the function of the PKD2 channel by physical interaction.The complex of PKD2 with TACAN, but not PKD2 alone, confers mechanosensitivity.