Phospholipase D2 localizes to the plasma membrane and regulates angiotensin II receptor endocytosis
Phospholipase D2 localizes to the plasma membrane and regulates angiotensin II receptor endocytosis
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DOI：
10.1091/mbc.e03-09-0673
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发表时间：
2004-03-01
影响因子：
3.3
通讯作者：
Frohman, MA
中科院分区：
文献类型：
--
作者：
Du, GW;Huang, P;Frohman, MA
磷脂酶 D (PLD) 是多种类型膜囊泡运输事件的关键促进剂。哺乳动物中存在两种 PLD 同工型：PLD1 和 PLD2。基于过表达研究的初步研究表明，在静息细胞中，人类 PLD1 主要定位于多种细胞类型的高尔基体和核周囊泡。相反，尽管在血清刺激后观察到膜囊泡的内化，但观察到过表达的小鼠 PLD2 主要定位于质膜。最近的一份报告表明，PLD2 到质膜的分配是错误的，因为对大鼠分泌细胞中的内源同工型进行成像并发现主要存在于高尔基体中。我们通过使用小鼠 PLD2 特异性单克隆抗体重新研究了这个问题，并发现，正如最初使用过表达研究报道的那样，内源性小鼠 PLD2 在多种细胞类型的质膜上最容易检测到。此外，我们报告称，小鼠、大鼠和人类 PLD2 在过表达时均类似地定位于所有三个物种细胞系的质膜上。最后，使用 PLD2 野生型活性或显性失活亚型的过表达以及 RNA 干扰介导的 PLD2 靶向进行的研究表明，PLD2 在质膜上发挥作用，促进血管紧张素 II 1 型受体的内吞作用。
Phospholipase D (PLD) is a key facilitator of multiple types of membrane vesicle trafficking events. Two PLD isoforms, PLD1 and PLD2, exist in mammals. Initial studies based on overexpression studies suggested that in resting cells, human PLD1 localized primarily to the Golgi and perinuclear vesicles in multiple cell types. In contrast, overexpressed mouse PLD2 was observed to localize primarily to the plasma membrane, although internalization on membrane vesicles was observed subsequent to serum stimulation. A recent report has suggested that the assignment of PLD2 to the plasma membrane is in error, because the endogenous isoform in rat secretory cells was imaged and found to be present primarily in the Golgi apparatus. We have reexamined this issue by using a monoclonal antibody specific for mouse PLD2, and find, as reported initially using overexpression studies, that endogenous mouse PLD2 is detected most readily at the plasma membrane in multiple cell types. In addition, we report that mouse, rat, and human PLD2 when overexpressed all similarly localize to the plasma membrane in cell lines from all three species. Finally, studies conducted using overexpression of wild-type active or dominant-negative isoforms of PLD2 and RNA interference-mediated targeting of PLD2 suggest that PLD2 functions at the plasma membrane to facilitate endocytosis of the angiotensin II type 1 receptor.