Differential involvement of ezrin/radixin/moesin proteins in sphingosine 1-phosphate-induced human pulmonary endothelial cell barrier enhancement.
Differential involvement of ezrin/radixin/moesin proteins in sphingosine 1-phosphate-induced human pulmonary endothelial cell barrier enhancement.
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DOI：
10.1016/j.cellsig.2011.08.003
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发表时间：
2011-12
影响因子：
4.8
通讯作者：
Dudek SM
中科院分区：
文献类型：
--
作者：
Adyshev DM;Moldobaeva NK;Elangovan VR;Garcia JG;Dudek SM
炎症激动剂诱导的内皮细胞屏障功能障碍是多种疾病中常见的病理生理事件。血小板衍生的磷脂鞘氨醇-1磷酸（S1 P）逆转这种功能障碍，通过一个涉及Rac GTP酶依赖性皮质肌动蛋白重排作为一个整体步骤的过程，有力地增强EC屏障。在这项研究中，我们探讨了肌动蛋白结合连接蛋白的埃兹蛋白、根蛋白和膜突蛋白（ERM）家族在调节S1 P诱导的人肺EC屏障增强中的作用。S1 P诱导ERM易位至EC外周并促进关键苏氨酸残基（Ezrin-567、Radixin-564、Moesin-558）上的ERM磷酸化。这种磷酸化依赖于PKC亚型和Rac 1的活化。S1 P后这些关键苏氨酸残基上的大多数ERM磷酸化发生在膜突蛋白和埃兹蛋白中。基线radixin磷酸化高于其他两种ERM蛋白，但在S1 P后不增加。S1 P诱导的膜突蛋白和埃兹蛋白苏氨酸磷酸化不是由屏障增强受体S1 PR 1介导的，因为S1 PR 1的siRNA下调不能抑制这些磷酸化事件，而用S1 PR 1特异性激动剂SEW 2871刺激EC不能诱导这些磷酸化事件。沉默所有的ERM蛋白或radixin单独（但不是膜突蛋白单独）减少S1 P诱导的Rac 1激活和下游Rac 1效应PAK 1的磷酸化。单独的Radixin siRNA或所有三种ERM蛋白的组合siRNA显著减弱S1 P诱导的EC屏障增强（通过跨内皮电阻（TER）、二磷酸-MLC的外周积累和皮质细胞骨架重排测量）。相反，膜突蛋白耗竭对这些参数有相反的影响。Ezrin沉默部分减弱S1 P诱导的EC屏障增强和细胞骨架变化因此，尽管结构上的相似性和报告的功能冗余，ERM蛋白差异调节S1 P诱导的肺EC细胞骨架和通透性的改变。这些结果表明，ERM激活是一个重要的调节事件EC屏障反应S1 P。
Endothelial cell (EC) barrier dysfunction induced by inflammatory agonists is a frequent pathophysiologic event in multiple diseases. The platelet-derived phospholipid sphingosine-1 phosphate (S1P) reverses this dysfunction by potently enhancing the EC barrier through a process involving Rac GTPase-dependent cortical actin rearrangement as an integral step. In this study we explored the role of the ezrin, radixin, and moesin (ERM) family of actin-binding linker protein in modulating S1P-induced human pulmonary EC barrier enhancement. S1P induces ERM translocation to the EC periphery and promotes ERM phosphorylation on a critical threonine residue (Ezrin-567, Radixin-564, Moesin-558). This phosphorylation is dependent on activation of PKC isoforms and Rac1. The majority of ERM phosphorylation on these critical threonine residues after S1P occurs in moesin and ezrin. Baseline radixin phosphorylation is higher than in the other two ERM proteins but does not increase after S1P. S1P-induced moesin and ezrin threonine phosphorylation is not mediated by the barrier enhancing receptor S1PR1 because siRNA downregulation of S1PR1 fails to inhibit these phosphorylation events, while stimulation of EC with the S1PR1-specific agonist SEW2871 fails to induce these phosphorylation events. Silencing of either all ERM proteins or radixin alone (but not moesin alone) reduced S1P-induced Rac1 activation and phosphorylation of the downstream Rac1 effector PAK1. Radixin siRNA alone, or combined siRNA for all three ERM proteins, dramatically attenuates S1P-induced EC barrier enhancement (measured by transendothelial electrical resistance (TER), peripheral accumulation of diphospho-MLC, and cortical cytoskeletal rearrangement. In contrast, moesin depletion has the opposite effects on these parameters. Ezrin silencing partially attenuates S1P-induced EC barrier enhancement and cytoskeletal changes. Thus, despite structural similarities and reported functional redundancy, the ERM proteins differentially modulate S1P-induced alterations in lung EC cytoskeleton and permeability. These results suggest that ERM activation is an important regulatory event in EC barrier responses to S1P.
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影响因子：
4.8
作者：
Canals, Daniel;Jenkins, Russell W.;Hannun, Yusuf A.
通讯作者：
Hannun, Yusuf A.
影响因子：
4.8
作者：
Gorshkova, Irina;He, Donghong;Natarajan, Viswanathan
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Natarajan, Viswanathan
磷酸肌醇的结合和磷酸化在ezrin的激活机理中依次起作用。
影响因子：
7.8
作者：
Fievet, BT;Gautreau, A;Roy, C;Del Maestro, L;Mangeat, P;Louvard, D;Arpin, M
通讯作者：
Arpin, M
影响因子：
4.8
作者：
Dudek, S. M.;Camp, S. M.;Singleton, P. A.
通讯作者：
Singleton, P. A.
影响因子：
4.8
作者：
Doi, Y;Itoh, M;Tsukita, S
通讯作者：
Tsukita, S