Roles for NHERF1 and NHERF2 on the regulation of C3a receptor signaling in human mast cells.
Roles for NHERF1 and NHERF2 on the regulation of C3a receptor signaling in human mast cells.
复制标题
DOI：
10.1371/journal.pone.0051355
复制
发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Ali H
中科院分区：
文献类型：
--
作者：
Subramanian H;Gupta K;Ali H
过敏毒素C3 a与G蛋白偶联受体（GPCR，C3 aR）结合，激活不同的信号通路，诱导人肥大细胞脱粒和细胞因子产生。衔接蛋白如Na+/H+交换调节因子（NHERF 1和NHERF 2）通过与存在于其胞质尾上的I类PDZ（PSD-95/Dlg/Zo 1）基序结合而参与调节某些GPCR的功能。虽然C3 aR具有I类PDZ基序，但其与NHERF蛋白相互作用以调节人肥大细胞中信号传导的可能性尚未确定。通过逆转录PCR和Western blotting，我们发现NHERF 1和NHERF 2在人肥大细胞系（HMC-1，LAD 2）和CD 34+来源的原代人肥大细胞中表达。然而，令人惊讶的是，C3 aR并不与这些衔接蛋白。为了评估NHERFs在C3 aR下游信号传导中的作用，我们使用慢病毒shRNA稳定敲低这些蛋白在人肥大细胞中的表达。沉默NHERF 1和NHERF 2的表达对C3 aR脱敏、激动剂诱导的受体内化、ERK/Akt磷酸化或趋化性没有影响。然而，NHERF 1和NHERF 2的缺失导致C3 a诱导的肥大细胞脱颗粒、NF-κB活化和趋化因子产生的显着抑制。这项研究表明，虽然C3 aR具有I类PDZ基序，但它不与NHERF 1和NHERF 2相关。令人惊讶的是，这些蛋白质为人肥大细胞中C3 a诱导的脱粒、NF-κB活化和趋化因子产生提供刺激信号。这些发现揭示了一个新的水平的复杂性的功能调节C3 aR的NHERFs在人类肥大细胞。
The anaphylatoxin C3a binds to the G protein coupled receptor (GPCR, C3aR) and activates divergent signaling pathways to induce degranulation and cytokine production in human mast cells. Adapter proteins such as the Na+/H+ exchange regulatory factor (NHERF1 and NHERF2) have been implicated in regulating functions of certain GPCRs by binding to the class I PDZ (PSD-95/Dlg/Zo1) motifs present on their cytoplasmic tails. Although C3aR possesses a class I PDZ motif, the possibility that it interacts with NHERF proteins to modulate signaling in human mast cells has not been determined. Using reverse transcription PCR and Western blotting, we found that NHERF1 and NHERF2 are expressed in human mast cell lines (HMC-1, LAD2) and CD34+-derived primary human mast cells. Surprisingly, however, C3aR did not associate with these adapter proteins. To assess the roles of NHERFs on signaling downstream of C3aR, we used lentiviral shRNA to stably knockdown the expression of these proteins in human mast cells. Silencing the expression of NHERF1 and NHERF2 had no effect on C3aR desensitization, agonist-induced receptor internalization, ERK/Akt phosphorylation or chemotaxis. However, loss of NHERF1 and NHERF2 resulted in significant inhibition of C3a-induced mast cell degranulation, NF-κB activation and chemokine production. This study demonstrates that although C3aR possesses a class I PDZ motif, it does not associate with NHERF1 and NHERF2. Surprisingly, these proteins provide stimulatory signals for C3a-induced degranulation, NF-κB activation and chemokine generation in human mast cells. These findings reveal a new level of complexity for the functional regulation of C3aR by NHERFs in human mast cells.
登录
查看更多内容
影响因子：
3.7
作者：
Gupta K;Subramanian H;Klos A;Ali H
通讯作者：
Ali H
影响因子：
4.4
作者：
Ali, H;Ahamed, J;Patel, DD
通讯作者：
Patel, DD
影响因子：
4.8
作者：
Mahon, MJ;Segre, GV
通讯作者：
Segre, GV
影响因子：
20.3
作者：
Hartmann, K;Henz, BM;Zuberbier, T
通讯作者：
Zuberbier, T
影响因子：
5.5
作者：
Capuano, Paola;Bacic, Desa;Murer, Heini
通讯作者：
Murer, Heini