The T1-tetramerisation domain of Kv1.2 rescues expression and preserves function of a truncated NaChBac sodium channel.
The T1-tetramerisation domain of Kv1.2 rescues expression and preserves function of a truncated NaChBac sodium channel.
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DOI：
10.1002/1873-3468.14279
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发表时间：
2022-03
期刊：
影响因子：
3.5
通讯作者：
O'Reilly, Andrias O.
中科院分区：
文献类型：
--
作者：
D'Avanzo, Nazzareno;Miles, Andrew J.;Powl, Andrew M.;Nichols, Colin G.;Wallace, B. A.;O'Reilly, Andrias O.
胞质结构域经常促进多聚体离子通道的功能组装。为了研究这一过程的结构决定因素，我们通过截短其C末端结构域并将Kv1.2通道的T1-四聚体结构域剪接到N末端来产生NaChBac钠通道的“T1-嵌合体”构建体。纯化的T1-嵌合体通道是四聚体，当重组到蛋白脂质体中时传导Na+，并被药物米贝拉地尔功能性阻断。T1嵌合体和全长NaChBac在膜中的表达水平相当，而缺乏胞质结构域的NaChBac突变体的膜表达大大降低。我们的研究结果支持了一个模型，使跨膜区域紧密接近，使他们的四聚化。这种现象与其他渠道，因此，我们的研究结果证实这是一个共同的组装机制。本研究探讨了细胞质结构域对离子通道组装的影响。当NaChBac细菌钠通道的C末端卷曲螺旋结构域与N末端上不相关的四聚体结构域交换时，嵌合体在膜中稳健表达并产生稳定的功能性四聚体通道。这表明，使跨膜区域接近促进NaChBac功能组装。
Cytoplasmic domains frequently promote functional assembly of multimeric ion channels. To investigate structural determinants of this process, we generated the ‘T1‐chimera’ construct of the NaChBac sodium channel by truncating its C‐terminal domain and splicing the T1‐tetramerisation domain of the Kv1.2 channel to the N terminus. Purified T1‐chimera channels were tetrameric, conducted Na+ when reconstituted into proteoliposomes, and were functionally blocked by the drug mibefradil. Both the T1‐chimera and full‐length NaChBac had comparable expression levels in the membrane, whereas a NaChBac mutant lacking a cytoplasmic domain had greatly reduced membrane expression. Our findings support a model whereby bringing the transmembrane regions into close proximity enables their tetramerisation. This phenomenon is found with other channels, and thus, our findings substantiate this as a common assembly mechanism. This study investigated the influence of the cytoplasmic domain on ion channel assembly. When the C‐terminal coiled‐coil domain of the NaChBac bacterial sodium channel was swapped with an unrelated tetramerisation domain on the N terminus, the chimera expressed robustly in the membrane and produced stable, functional tetrameric channels. This indicates that bringing the transmembrane regions into proximity promotes NaChBac functional assembly.
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影响因子：
3.7
作者：
D'Avanzo N;McCusker EC;Powl AM;Miles AJ;Nichols CG;Wallace BA
通讯作者：
Wallace BA
影响因子：
5.6
作者：
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通讯作者：
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影响因子：
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通讯作者：
Wallace, B. A.
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Clapham DE
影响因子：
56.9
作者：
HOSHI, T;ZAGOTTA, WN;ALDRICH, RW
通讯作者：
ALDRICH, RW