Effect of inhibition of tyrosine phosphatases on voltage‐operated calcium channel currents in rabbit isolated ear artery cells

Effect of inhibition of tyrosine phosphatases on voltage‐operated calcium channel currents in rabbit isolated ear artery cells
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酪氨酸磷酸酶抑制对兔离体耳动脉细胞电压驱动钙通道电流的影响

DOI:
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发表时间:
1998
影响因子:
7.3
通讯作者:
A. Hughes
A. Hughes
中科院分区:
医学2区
文献类型:
--
作者:
S. Wijetunge;J. Lymn;A. Hughes

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通过抑制内源性酪氨酸磷酸酶增加细胞酪氨酸磷酸化,研究了血管平滑肌细胞电压操作钙通道电流的影响。在兔单耳动脉平滑肌细胞中,全细胞电压钳技术研究发现,酪氨酸磷酸酶抑制剂、正钒酸钠(100 μM)和过钒酸钠(100 μM +1 mM H2O2)在细胞内分别使电压操作的钙通道电流增加56%和83%。另外两种膜渗透酪氨酸磷酸酶抑制剂,苯larsin oxide (100 μM)和dephostatin (50 μM)也分别使电压操作的钙通道电流增加48%和52%。选择性酪氨酸激酶抑制剂tyrphostin‐23 (100 μM)可使钙通道电流降低41%。用tyrphostin - 23预孵育可消除过氧钒酸盐、氧化苯larsine和去光抑素对钙通道的影响。兔耳动脉细胞裂解物的Western blot分析显示,经过过氧钒酸盐处理后,几种内源性蛋白的酪氨酸磷酸化增加。这些结果表明,一些结构不同的酪氨酸磷酸酶抑制剂增加了动脉平滑肌细胞中电压操作的钙通道电流,可能是由于酪氨酸磷酸化增加。
The effect of increasing cellular tyrosine phosphorylation by inhibiting endogenous tyrosine phosphatases was examined on voltage‐operated calcium channel currents in vascular smooth muscle cells. In single ear artery smooth muscle cells of the rabbit, studied by the whole cell voltage clamp technique, intracellular application of the tyrosine phosphatase inhibitors, sodium orthovanadate (100 μM) and peroxyvanadate (100 μM orthovanadate+1 mM H2O2) increased voltage‐operated calcium channel currents by 56% and 83%, respectively. Bath application of two other membrane permeant tyrosine phosphatase inhibitors, phenylarsine oxide (100 μM) and dephostatin (50 μM) also increased voltage‐operated calcium channel currents by 48% and 52%, respectively. The selective tyrosine kinase inhibitor, tyrphostin‐23 (100 μM) reduced calcium channel currents by 41%. Pre‐incubation with tyrphostin‐23 abolished the effects of peroxyvanadate, phenylarsine oxide and dephostatin on calcium channels. Western blot analysis of rabbit ear artery cell lysates showed increased tyrosine phosphorylation of several endogenous proteins following treatment with peroxyvanadate. These results indicate that a number of structurally dissimilar inhibitors of tyrosine phosphatases increase voltage‐operated calcium channel currents in arterial smooth muscle cells presumably due to increased tyrosine phosphorylation.
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