Protein kinase C isozyme-specific potentiation of expressed Ca v 2.3 currents by acetyl-beta-methylcholine and phorbol-12-myristate, 13-acetate.
Protein kinase C isozyme-specific potentiation of expressed Ca v 2.3 currents by acetyl-beta-methylcholine and phorbol-12-myristate, 13-acetate.
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乙酰-β-甲基胆碱和佛波醇-12-肉豆蔻酸酯、13-乙酸酯对表达的 Ca v 2.3 电流的蛋白激酶 C 同工酶特异性增强。
DOI：
10.1016/j.brainres.2008.03.017
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发表时间：
2008
期刊：
影响因子：
2.9
通讯作者：
Kamatchi,GanesanL
中科院分区：
文献类型：
--
作者：
Rajagopal,Senthilkumar;Fang,Hongyu;Patanavanich,Saharat;Sando,JulianneJ;Kamatchi,GanesanL
蛋白激酶 C (PKC) 与乙酰-β-甲基胆碱 (MCh)（一种毒蕈碱 M1 受体激动剂或佛波醇 12-肉豆蔻酸酯、13-乙酸酯 (PMA)）对 Cav2.3 电流的增强有关。使用易位作为激活的量度并使用同工酶选择性拮抗剂和 siRNA 研究了负责 MCh 和 PMA 作用的 PKC 同工酶。爪蟾卵母细胞中的 α12.3、β1b 和 α2δ 亚基和毒蕈碱 M1 受体表达腔通道，并使用 Ba2+ 作为电荷载体研究表达电流 (IBa)。通过蛋白质印迹研究 PKC 同工酶向膜的易位表明，所有 11 种已知的 PKC 同工酶都存在于非洲爪蟾卵母细胞中。卵母细胞暴露于 MCh 导致 PKC α 易位，而 PMA 激活 PKC βII 和 ɛ 同工酶。 MCh 的作用受到 Go 6976（cPKC 同工酶抑制剂或 PKC α siRNA）的抑制。 PMA 诱导的 Cav2.3 电流增强被 CG533 53（一种 PKC βII 拮抗剂）、βIIV5.3（一种 PKC βII 的肽易位抑制剂）或 PKC βII siRNA 抑制。同样，PKC ɛ 的肽易位抑制剂 ɛV1.2 或 PKC ɛ siRNA 抑制 PMA 作用。 PKC抑制剂轻微增加基础IB。某些 PKC 同工酶可能对 IBa 具有阴性控制作用。我们的结果表明 PKC α 涉及 MCh 对 Cav2.3 电流的增强，而 PKC βII 和 ɛ 涉及 PMA 对 Cav2.3 电流的增强。
Protein kinase C (PKC) is implicated in the potentiation of Cav2.3 currents by acetyl-β-methylcholine (MCh), a muscarinic M1receptor agonist or phorbol-12-myristate, 13-acetate (PMA). The PKC isozymes responsible for the action of MCh and PMA were investigated using translocation as a measure of activation and with isozyme-selective antagonists and siRNA. Cavchannels were expressed with α12.3, β1b and α2δ subunits and muscarinic M1receptors in the Xenopus oocytes and the expressed currents (IBa) were studied using Ba2+as the charge carrier. Translocation of PKC isozymes to the membrane studied by Western blot revealed that all eleven known PKC isozymes are present in the Xenopus oocytes. Exposure of the oocytes to MCh led to the translocation of PKC α whereas PMA activated PKC βII and ɛ isozymes. The action of MCh was inhibited by Go 6976, an inhibitor of cPKC isozymes or PKC α siRNA. PMA-induced potentiation of Cav2.3 currents was inhibited by CG533 53, a PKC βII antagonist, βIIV5.3, a peptide translocation inhibitor of PKC βII or PKC βII siRNA. Similarly, ɛV1.2, a peptide translocation inhibitor of PKC ɛ or PKC ɛ siRNA inhibited PMA action. The inhibitors of PKC increased the basal IBaslightly. It is possible that some PKC isozymes have negative control over the IBa. Our results implicate PKC α in the potentiation of Cav2.3 currents by MCh and PKC βII and ɛ in the potentiation of Cav2.3 currents by PMA.