Phosphatidylinositol 3-kinases regulate ERK and p38 MAP kinases in canine colonic smooth muscle.
Phosphatidylinositol 3-kinases regulate ERK and p38 MAP kinases in canine colonic smooth muscle.
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DOI：
10.1152/ajpcell.2000.279.2.c352
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发表时间：
2000-08
期刊：
影响因子：
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通讯作者：
I. Yamboliev;Kevin M. Wiesmann;C. Singer;J. Hedges;W. Gerthoffer
中科院分区：
文献类型：
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作者：
I. Yamboliev;Kevin M. Wiesmann;C. Singer;J. Hedges;W. Gerthoffer
在犬结肠中，M2/M3毒蕈碱受体与细胞外信号调节激酶（ERK）和p38丝裂原活化蛋白（MAP）激酶偶联。我们测试的假设，这种耦合介导的磷脂酰肌醇（PI）3-激酶家族的酶。RT-PCR和Western blotting证实了PI 3-激酶-α和PI 3-激酶-γ两种亚型的表达。完整肌条的毒蕈碱刺激（10 μ M ACh）激活PI 3-激酶-γ，ERK和p38 MAP激酶，以及MAP激酶激活的蛋白激酶-2，而PI 3-激酶-α激活未检测到。Wortmannin（25 μ M）消除PI 3-激酶-γ、ERK和p38 MAP激酶的激活。MAP激酶抑制是PI 3-激酶-γ-特异性效应，因为渥曼青霉素不抑制重组活化的鼠ERK 2 MAP激酶、蛋白激酶C、Raf-1或MAP激酶激酶。在培养的肌肉细胞中，新生小牛血清（3%）激活PI 3-激酶-α和PI 3-激酶-γ亚型，ERK和p38 MAP激酶，并刺激趋化细胞迁移。使用渥曼青霉素和LY-294002抑制PI 3-激酶活性，PD-098059和SB-203580抑制ERK和p38 MAP激酶，我们确定了这些酶在调节结肠肌细胞趋化性迁移中具有重要的功能。
In canine colon, M2/M3 muscarinic receptors are coupled to extracellular signal-regulated kinase (ERK) and p38 mitogen-activated protein (MAP) kinases. We tested the hypothesis that this coupling is mediated by enzymes of the phosphatidylinositol (PI) 3-kinase family. RT-PCR and Western blotting demonstrated expression of two isoforms, PI 3-kinase-alpha and PI 3-kinase-gamma. Muscarinic stimulation of intact muscle strips (10 microM ACh) activated PI 3-kinase-gamma, ERK and p38 MAP kinases, and MAP kinase-activated protein kinase-2, whereas PI 3-kinase-alpha activation was not detected. Wortmannin (25 microM) abolished the activation of PI 3-kinase-gamma, ERK, and p38 MAP kinases. MAP kinase inhibition was a PI 3-kinase-gamma-specific effect, since wortmannin did not inhibit recombinant activated murine ERK2 MAP kinase, protein kinase C, Raf-1, or MAP kinase kinase. In cultured muscle cells, newborn calf serum (3%) activated PI 3-kinase-alpha and PI 3-kinase-gamma isoforms, ERK and p38 MAP kinases, and stimulated chemotactic cell migration. Using wortmannin and LY-294002 to inhibit PI 3-kinase activity and PD-098059 and SB-203580 to inhibit ERK and p38 MAP kinases, we established that these enzymes are functionally important for regulation of chemotactic migration of colonic myocytes.