Apical localization of PMCA2w/b is lipid raft-dependent.

Apical localization of PMCA2w/b is lipid raft-dependent.
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PMCA2w/b 的顶端定位依赖于脂筏。

DOI:
10.1016/j.bbrc.2009.04.044
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发表时间:
2009
影响因子:
3.1
通讯作者:
Strehler,EmanuelE
Strehler,EmanuelE
中科院分区:
生物学4区
文献类型:
--
作者:
Xiong,Yuning;Antalffy,Géza;Enyedi,Agnes;Strehler,EmanuelE

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MDCK细胞中第一胞内环的选择性剪接差异靶向质膜钙ATP酶(PMCA)亚型2至顶膜或基底外侧膜。为了确定靶向是否受脂质相互作用的影响,我们在MDCK细胞中稳定表达PMCA 2 w/B和PMCA 2 z/B,并通过共聚焦荧光显微镜和膜分级分离分析PMCA分布。PMCA 2 w/B在基底膜和侧膜上均有分布,而PMCA 2 z/B主要分布于基底膜。PMCA 2 w/B的一个重要部分分配到低密度膜与脂筏。甲基-β-环糊精对膜胆固醇的消耗导致脂筏结合减少,顶膜PMCA 2 w/B显著丢失,而PMCA 2 z/B的侧向定位保持不变。我们的数据表明,选择性剪接不同地影响PMCA 2 w/B和PMCA 2 z/B的脂质相互作用,并且PMCA 2 w/B的顶端定位是脂筏依赖性的,并且对胆固醇消耗敏感。
Alternative splicing of the first intracellular loop differentially targets plasma membrane calcium ATPase (PMCA) isoform 2 to the apical or basolateral membrane in MDCK cells. To determine if the targeting is affected by lipid interactions, we stably expressed PMCA2w/b and PMCA2z/b in MDCK cells, and analyzed the PMCA distribution by confocal fluorescence microscopy and membrane fractionation. PMCA2w/b showed clear apical and lateral distribution, whereas PMCA2z/b was mainly localized to the basolateral membrane. A significant fraction of PMCA2w/b partitioned into low-density membranes associated with lipid rafts. Depletion of membrane cholesterol by methyl-β-cyclodextrin resulted in reduced lipid raft association and a striking loss of PMCA2w/b from the apical membrane, whereas the lateral localization of PMCA2z/b remained unchanged. Our data indicate that alternative splicing differentially affects the lipid interactions of PMCA2w/b and PMCA2z/b and that the apical localization of PMCA2w/b is lipid raft-dependent and sensitive to cholesterol depletion.
胆固醇消耗降低了上皮 Madin-Darby 犬肾细胞的顶端转运能力。
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