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首页> 外文期刊>Journal of Cellular Physiology >Dynamic redistribution of calcium sensitive potassium channels (hK(Ca)3.1) in migrating cells.
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Dynamic redistribution of calcium sensitive potassium channels (hK(Ca)3.1) in migrating cells.

机译：动态分配的钙敏感钾离子通道在迁移((Ca) 3.1港元)细胞。

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Calcium-sensitive potassium channels (K(Ca)3.1) are expressed in virtually all migrating cells. Their activity is required for optimal cell migration so that their blockade leads to slowing down. K(Ca)3.1 channels must be inserted into the plasma membrane in order to elicit their physiological function. However, the plasma membrane of migrating cells is subject to rapid recycling by means of endo- and exocytosis. Here, we focussed on the endocytic internalization and the intracellular transport of the human isoform hK(Ca)3.1. A hK(Ca)3.1 channel construct with an HA-tag in the extracellularly located S3-S4 linker was transfected into migrating transformed renal epithelial MDCK-F cells. Channel internalization was visualized and quantified with immunofluorescence and a cell-based ELISA. Movement of hK(Ca)3.1 channel containing vesicles as well as migration of MDCK-F cells were monitored by means of time lapse video microscopy. hK(Ca)3.1 channels are endocytosed during migration. Most of the hK(Ca)3.1 channel containing vesicles are moving at a speed of up to 2 microm/sec in a microtubule-dependent manner towards the front of MDCK-F cells. Our experiments indicate that endocytosis of hK(Ca)3.1 channels is clathrin-dependent since they colocalize with clathrin adaptor proteins and since it is impaired when a C-terminal dileucine motif is mutated. The C-terminal dileucine motif is also important for the subcellular localization of hK(Ca)3.1 channels in migrating cells. Mutated channels are no longer concentrated at the leading edge. We therefore propose that recycling of hK(Ca)3.1 channels contributes to their characteristic subcellular distribution in migrating cells.
机译：Calcium-sensitive钾离子通道(3.1 K (Ca))在几乎所有的迁移细胞表达。他们的活动需要最佳的细胞迁移,这样他们的封锁导致放缓下来。等离子体膜以引起生理功能。膜移植细胞的快速回收通过endo -胞外分泌。我们关注的内吞作用的内化人类同种型的胞内运输(Ca) 3.1港元。送到位于S3-S4 HA-tag链接器被转染进迁移转化肾上皮MDCK-F细胞。内化是可视化和量化免疫荧光和细胞ELISA。运动包含囊泡(Ca) 3.1港元的通道以及MDCK-F细胞迁移通过时间推移视频监控显微镜。在迁移。含有囊泡运动的速度2 microm microtubule-dependent方式/秒向前面MDCK-F细胞。实验表明,内吞作用(Ca) 3.1港元clathrin-dependent因为渠道他们用网格蛋白colocalize适配器蛋白质,因为它是当一个c端受损dileucine主题是突变。dileucine主题也很重要的亚细胞定位(Ca) 3.1港元的渠道迁移细胞。集中在前缘。建议(Ca) 3.1港元的回收渠道有助于他们的亚细胞的特征在迁移细胞分布。

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