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首页> 外文期刊>International Journal of Cell Biology >Impact of Hybrid and Complex N-Glycans on Cell Surface Targeting of the Endogenous Chloride Cotransporter Slc12a2
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Impact of Hybrid and Complex N-Glycans on Cell Surface Targeting of the Endogenous Chloride Cotransporter Slc12a2

机译：杂种和复杂的N-聚糖对内源性氯化物共转运蛋白Slc12a2的细胞表面靶向的影响

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摘要

The Na~+K~+2Cl~? cotransporter-1 (Slc12a2, NKCC1) is widely distributed and involved in cell volume/ion regulation. Functional NKCC1 locates in the plasma membrane of all cells studied, particularly in the basolateral membrane of most polarized cells. Although the mechanisms involved in plasma membrane sorting of NKCC1 are poorly understood, it is assumed that Nglycosylation is necessary. Here, we characterize expression, N-glycosylation, and distribution of NKCC1 in COS7 cells. We show that ~25% of NKCC1 is complex N-glycosylated whereas the rest of it corresponds to core/high-mannose and hybrid-type Nglycosylated forms. Further, ~10% of NKCC1 reaches the plasma membrane, mostly as core/high-mannose type, whereas ~90% of NKCC1 is distributed in defined intracellular compartments. In addition, inhibition of the first step of N-glycan biosynthesis with tunicamycin decreases total and plasma membrane located NKCC1 resulting in almost undetectable cotransport function. Moreover, inhibition of N-glycan maturation with swainsonine or kifunensine increased core/hybrid-type NKCC1 expression but eliminated plasma membrane complex N-glycosylated NKCC1 and transport function. Together, these results suggest that (i) NKCC1 is delivered to the plasmamembrane of COS7 cells independently of its N-glycan nature, (ii) most of NKCC1 in the plasma membrane is core/hybrid-type N-glycosylated, and (iii) the minimal proportion of complex N-glycosylated NKCC1 is functionally active.
机译：Na〜+ K〜+ 2Cl〜？ cotransporter-1（Slc12a2，NKCC1）广泛分布并参与细胞体积/离子调节。功能性NKCC1位于所有研究细胞的质膜中，特别是在大多数极化细胞的基底外侧膜中。尽管与NKCC1的质膜分选有关的机制了解甚少，但可以认为N糖基化是必要的。在这里，我们表征NK7在COS7细胞中的表达，N-糖基化和分布。我们显示〜25％的NKCC1是复杂的N-糖基化的，而其余的则对应于核心/高甘露糖和杂合型N糖基化的形式。此外，约10％的NKCC1到达质膜，主要是核心/高甘露糖型，而约90％的NKCC1分布在确定的细胞内区室中。另外，衣霉素抑制了N-聚糖生物合成的第一步，减少了总的和定位于NKCC1的质膜，导致几乎不可检测的共转运功能。此外，用swainsonine或kifunensine抑制N-聚糖成熟可增加核心/杂合型NKCC1表达，但消除了质膜复合物N-糖基化的NKCC1和转运功能。在一起，这些结果表明（i）NKCC1被传递到COS7细胞的质膜，而与其N-聚糖性质无关，（ii）质膜中的大部分NKCC1是核心/杂合型N-糖基化的，和（iii）最小比例的复合N-糖基化NKCC1具有功能活性。

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