N-glycosylation–dependent regulation of hK2P17.1 currents
N-glycosylation–dependent regulation of hK2P17.1 currents
复制标题
hK2P17 1 电流的 N-糖基化依赖性调节
DOI：
10.1091/mbc.e18-10-0687
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发表时间：
2019
影响因子：
3.3
通讯作者：
Schmidt C
中科院分区：
文献类型：
--
作者：
Wiedmann F;Schlund D;Voigt N;Ratte A;Kraft M;Katus HA;Schmidt C
两个孔域钾(K2P)通道介导钾背景电流，稳定静息膜电位，促进动作电位复极化。在人类心脏中，hK2P17.1通道主要在心房和浦肯野细胞中表达。心房颤动或心力衰竭患者的心房hK2P17.1蛋白水平降低。HK2P17.1基因改变与心脏传导障碍相关。目前对hK2P17.1的翻译后修饰知之甚少。在这里，我们对hK2P17.1的糖基化进行了表征，并研究了糖基化如何改变其表面表达和活性。在非洲爪哇细胞、HEK-293T细胞和HeLa细胞中表达野生型hK2P17.1通道和缺乏特定糖基化位点的通道。N-糖苷酶F和衣霉素阻断N-糖基化。免疫印迹分析和双电极电压钳技术检测hK2P17.1的表达和活性。HK2P17.1的通道亚基在N65和N94处含有两个功能性N-糖基化位点。在半糖基化的hK2P17.1通道中，功能和膜运输保持不变。这两个N-糖基化位点的破坏会导致hK2P17.1电流的丢失，这可能是由于表面表达受损引起的。本研究证实了hK2P17.1通道亚基的二糖基化及其在细胞表面靶向中的关键作用。我们的发现强调了N-糖基化在离子通道的生物发生和膜运输中的功能相关性。
Two pore-domain potassium (K2P) channels mediate potassium background currents that stabilize the resting membrane potential and facilitate action potential repolarization. In the human heart, hK2P17.1 channels are predominantly expressed in the atria and Purkinje cells. Reduced atrial hK2P17.1 protein levels were described in patients with atrial fibrillation or heart failure. Genetic alterations in hK2P17.1 were associated with cardiac conduction disorders. Little is known about posttranslational modifications of hK2P17.1. Here, we characterized glycosylation of hK2P17.1 and investigated how glycosylation alters its surface expression and activity. Wild-type hK2P17.1 channels and channels lacking specific glycosylation sites were expressed inXenopus laevisoocytes, HEK-293T cells, and HeLa cells. N-glycosylation was disrupted using N-glycosidase F and tunicamycin. hK2P17.1 expression and activity were assessed using immunoblot analysis and a two-electrode voltage clamp technique. Channel subunits of hK2P17.1 harbor two functional N-glycosylation sites at positions N65 and N94. In hemi-glycosylated hK2P17.1 channels, functionality and membrane trafficking remain preserved. Disruption of both N-glycosylation sites results in loss of hK2P17.1 currents, presumably caused by impaired surface expression. This study confirms diglycosylation of hK2P17.1 channel subunits and its pivotal role in cell-surface targeting. Our findings underline the functional relevance of N-glycosylation in biogenesis and membrane trafficking of ion channels.
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影响因子：
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发表时间：
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影响因子：
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