Epidermal growth factor receptor activation of calpain is required for fibroblast motility and occurs via an ERK/MAP kinase signaling pathway
Epidermal growth factor receptor activation of calpain is required for fibroblast motility and occurs via an ERK/MAP kinase signaling pathway
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DOI：
10.1074/jbc.275.4.2390
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发表时间：
2000-01-28
影响因子：
4.8
通讯作者：
Wells, A
中科院分区：
文献类型：
--
作者：
Glading, A;Chang, P;Wells, A
为了迁移，细胞必须重新组织它们与基质的连接，并且在运动过程中它们必须打破后部连接。这些生物物理过程背后的分子和生物化学机制尚不清楚。最近的研究表明，细胞外信号调节激酶/促分裂原活化蛋白（ERK/MAP）激酶和钙蛋白酶（EC 3.4.22.17）参与了这些过程，但尚不确定这是否是两种不同的途径，作用于不同的运动模式。我们报道了表皮生长因子（EGF）受体介导的成纤维细胞运动中涉及的细胞去粘附需要ERK/MAP激酶信号下游的M-钙蛋白酶的激活，表达全长轻度型表皮生长因子受体的NR 6成纤维细胞需要钙蛋白酶和ERK的激活，如药理学抑制剂所证明的（分别为钙蛋白酶和钙蛋白酶抑制剂I和PD 98059）用于EGF诱导的去粘附和运动。EGF诱导的钙蛋白酶的快速激活，这是可预防的分子抑制的Ras-Raf-MEK，但不是磷脂酶C γ信号通路，钙蛋白酶的刺激转染的组成型活性MEK。钙蛋白酶活性的增强并不反映钙蛋白酶蛋白水平的增加或其内源性抑制剂钙蛋白酶抑制素水平的降低。ERK/MAP激酶信号传导和细胞运动之间的联系需要钙蛋白酶的M-亚型（钙蛋白酶II），如通过特异性反义介导的下调所确定的。这些数据促进了先前未描述的ERK/MAP激酶激活钙蛋白酶的信号传导途径，以响应于EGF刺激而使细胞-基质粘附不稳定。
To become migratory, cells must reorganize their connections to the substratum, and during locomotion they must break rear attachments. The molecular and biochemical mechanisms underlying these biophysical processes are unknown. Recent studies have implicated both extracellular signal-regulated kinase/mitogen-activated protein (ERK/MAP) kinase and calpain (EC 3.4.22.17) in these processes, but it is uncertain whether these are two distinct pathways acting on different modes of motility. We report that cell deadhesion involved in epidermal growth factor (EGF) receptor-mediated fibroblast motility requires activation of M-calpain downstream of ERK/MAP kinase signaling, NR6 fibroblasts expressing full-length mild type epidermal growth factor receptor required both calpain and ERK activation, as demonstrated by pharmacological inhibitors (calpeptin and calpain inhibitor I and PD98059, respectively) for EGF-induced deadhesion and motility. EGF induced rapid activation of calpain that was preventable by molecular inhibition of the Ras-Raf-MEK but not phospholipase C gamma signaling pathway, and calpain was stimulated by transfection of constitutively active MEK. Enhanced calpain activity was not mirrored by increased calpain protein levels or decreased levels of its endogenous inhibitor calpastatin. The link between ERK/MAP kinase signaling and cell motility required the M-isoform of calpain (calpain II), as determined by specific antisense-mediated down-regulation, These data promote a previously undescribed signaling pathway of ERK/MAP kinases activating calpain to destabilize cell-substratum adhesions in response to EGF stimulation.