Phosphorylation of epidermal growth factor receptor at serine 1047 in cultured lung alveolar epithelial cells by bradykinin B2 receptor stimulation
Phosphorylation of epidermal growth factor receptor at serine 1047 in cultured lung alveolar epithelial cells by bradykinin B2 receptor stimulation
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DOI：
10.1016/j.pupt.2017.09.002
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发表时间：
2018-02-01
影响因子：
3.2
通讯作者：
Yamamoto, Hideyuki
中科院分区：
文献类型：
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作者：
Izumi, Shunsuke;Higa-Nakamine, Sayomi;Yamamoto, Hideyuki
越来越多的证据表明，表皮生长因子受体（EGFR）通过丝氨酸1047（Ser 1047）的磷酸化而脱敏。我们和其他研究小组已经报道，在培养的肺泡上皮A549细胞中，肿瘤坏死因子α（TNF α）和Toll样受体5（TLR 5）的受体的刺激通过激活p38丝裂原活化蛋白激酶（p38 MAPK）诱导Ser 1047的磷酸化。然而，在任何培养的细胞中，通过刺激任何G-蛋白偶联受体（GPCR）使EGFR在Ser 1047处磷酸化尚未报道。在本研究中，我们首先证实，A549细胞表达缓激肽（BK）B2受体，然后，我们检查是否BK处理A549细胞激活MAPK和诱导EGFR的Ser 1047磷酸化。免疫印迹分析和报告基因分析表明，BK激活细胞外信号调节激酶（ERK）和p38 MAPK途径。抑制剂研究表明G（q/11）主要参与ERK和p38 MAPK的激活。我们发现，BK B2受体的刺激，而不是BK B1受体，诱导EGFR的丝氨酸1047磷酸化。药理实验表明ERK和p38 MAPK均参与EGFR的磷酸化。这些结果有力地表明BK调节肺泡上皮细胞中EGFR的功能。此外，BK处理后，ERK依赖性地增加了细胞内双特异性MAPK磷酸酶5（DUSP 5）的mRNA水平，提示ERK在细胞内存在负反馈机制。(C)2017爱思唯尔有限公司版权所有
Accumulating evidence indicates that epidermal growth factor receptor (EGFR) is desensitized by phosphorylation of serine 1047 (Ser1047). We and other groups have reported that stimulation of a receptor of tumor-necrosis factor alpha (TNF alpha) and Toll-like receptor 5 (TLR5) induced the phosphorylation of Ser1047 through activation of p38 mitogen-activated protein kinase (p38 MAPK) in cultured lung alveolar epithelial A549 cells. However, phosphorylation of EGFR at Ser1047 by stimulation of any G-protein coupled receptors (GPCRs) has not been reported in any cultured cells. In the present study, we first confirmed that A549 cells expressed bradykinin (BK) B2 receptor, and then, we examined whether BK treatment of A549 cells activated MAPKs and induced the phosphorylation of EGFR at Ser1047. Immunoblotting analysis and reporter gene assays indicated that BK activated the pathways of extracellular signal-regulated kinase (ERK) and p38 MAPK. Inhibitor studies suggested that G(q/11) was mainly involved in the activation of ERK and p38 MAPK. We found that stimulation of the BK B2 receptor, but not the BK B1 receptor, induced phosphorylation of EGFR at Ser1047. Pharmacological experiments indicated that both ERK and p38 MAPK were involved in the phosphorylation of EGFR. These results strongly suggested that BK regulates EGFR functions in lung alveolar epithelial cells. In addition, we found that BK treatment increased the mRNA level of dual specificity MAPK phosphatase 5 (DUSP5) in an ERIC-dependent manner, which suggested that a negative feedback mechanism of ERK existed in the cells. (C) 2017 Elsevier Ltd. All rights reserved.