Role of Tyrosine Phosphorylation in Potassium Channel Activation
Role of Tyrosine Phosphorylation in Potassium Channel Activation
复制标题
酪氨酸磷酸化在钾通道激活中的作用
DOI：
10.1074/jbc.270.41.24292
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发表时间：
1995
期刊：
影响因子：
--
通讯作者：
B. Dufy
中科院分区：
文献类型：
--
作者：
N. Prevarskaya;R. Skryma;P. Vacher;N. Daniel;J. Djiane;B. Dufy
用长形式的催乳素 (PRL) 受体 (PRL-R) cDNA 稳定转染的中国仓鼠卵巢 (CHO) 细胞用于 PRL-R 信号转导研究。采用全细胞和无细胞配置的膜片钳技术。将转染的 CHO 细胞暴露于 5 nM PRL 会导致 Ca2+- 和电压依赖性 K+ 通道 (KCa) 活性增加。这种效果是直接的，因为在切除贴片实验中也观察到了这一点。研究了一系列酪氨酸激酶抑制剂，以研究蛋白酪氨酸激酶可能参与 KCa 功能及其受 PRL 的刺激。金雀异黄素、薰衣草素 A 和除草霉素 A 以浓度和时间依赖性方式降低全细胞中 KCa 电流的幅度以及无细胞实验中 KCa 通道的开放概率。随后应用PRL无效。蛋白酪氨酸磷酸酶抑制剂原钒酸盐 (1 mM) 刺激切除斑块中的 KCa 通道活性，表明蛋白酪氨酸激酶和蛋白酪氨酸磷酸酶可以沿相反方向调节通道。此外，在全细胞实验以及切除的斑片记录中，抗 JAK2 酪氨酸激酶抗体降低了 KCa 电导和 KCa 通道的开放概率。随后应用 PRL 不再能够刺激 KCa 电导。使用相同的抗 JAK2 抗体的免疫印迹研究揭示了 JAK2 激酶与 PRL-R 的组成型关联。抗 JAK2 抗体与 JAK2 免疫肽预孵育消除了单独使用抗 JAK2 抗体在电生理学和免疫印迹研究中观察到的效果。我们从这些发现中得出结论，这些 KCa 通道是通过酪氨酸磷酸化/去磷酸化进行调节的； JAK2 酪氨酸激酶与 PRL-R 组成型相关，参与 KCa 通道的 PRL 刺激。
Chinese hamster ovary (CHO) cells, stably transfected with the long form of the prolactin (PRL) receptor (PRL-R) cDNA, were used for PRL-R signal transduction studies. Patch-clamp technique in whole cell and cell-free configurations were employed. Exposure of transfected CHO cells to 5 nM PRL led to the increase of Ca2+- and voltage-dependent K+ channel (KCa) activity. The effect was direct as it was observed also in excised patch experiments. A series of tyrosine kinase inhibitors was studied to investigate the possible involvement of protein tyrosine kinases in KCa functioning and its stimulation by PRL. Genistein, lavendustin A, and herbimycin A decreased in a concentration and time-dependent manner the amplitude of the KCa current in whole cell and the open probability of KCa channels in cell-free experiments. The subsequent application of PRL was ineffective. The protein tyrosine phosphatase inhibitor orthovanadate (1 mM) stimulated KCa channel activity in excised patches, indicating that channels can be modulated in opposite directions by protein tyrosine kinase and protein tyrosine phosphatase. Moreover, in whole cell experiments as well as in excised patch recordings, anti-JAK2 tyrosine kinase antibody decreased the KCa conductance and the open probability of the KCa channels. Subsequent application of PRL was no longer able to stimulate KCa conductance. Immunoblotting studies using the same anti-JAK2 antibody, revealed the constitutive association of JAK2 kinase with PRL-R. Preincubation of anti-JAK2 antibody with the JAK2 Immunizing Peptide abolished the effects observed using anti-JAK2 antibody alone in both electrophysiological and immunoblotting studies. We conclude from these findings that these KCa channels are regulated through tyrosine phosphorylation/dephosphorylation; JAK2 tyrosine kinase, constitutively associated with PRL-R, is implicated in PRL stimulation of KCa channels.
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DOI：
10.1073/pnas.85.19.7182
发表时间：
1988-10-01
影响因子：
11.1
作者：
CHARBONNEAU, H;TONKS, NK;FISCHER, EH
通讯作者：
FISCHER, EH
生长激素受体相关酪氨酸激酶参与生长激素作用的证据。
DOI：
--
发表时间：
1993
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Campbell,GS;Christian,LJ;Carter-Su,C
通讯作者：
Carter-Su,C
催乳素激活受体相关酪氨酸激酶 JAK2。
DOI：
--
发表时间：
1994
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Rui,H;Kirken,RA;Farrar,WL
通讯作者：
Farrar,WL
钒酸盐抑制膜磷酸酪氨酰蛋白磷酸酶活性。
DOI：
10.1016/0006-291x(82)90635-0
发表时间：
1982
影响因子：
3.1
作者：
Swarup,G;Cohen,S;Garbers,DL
通讯作者：
Garbers,DL