Nonspecific, reversible inhibition of voltage-gated calcium channels by CaMKII inhibitor CK59.
Nonspecific, reversible inhibition of voltage-gated calcium channels by CaMKII inhibitor CK59.
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DOI：
10.1007/s10571-013-9941-8
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发表时间：
2013-07
影响因子：
4
通讯作者：
Mynlieff, Michelle
中科院分区：
文献类型：
--
作者：
Karls, Andrew S.;Mynlieff, Michelle
关键词：
激酶相关过程的研究通常使用药理学抑制来揭示激酶参与的途径。然而，使用此类激酶抑制剂的一个问题是它们可能缺乏特异性。在这里，我们报告，钙-钙调蛋白依赖性激酶II（CaMKII）抑制剂CK 59抑制多种电压门控钙通道，包括L型通道在去极化的剂量依赖性方式。使用另一种CaMKII抑制剂，细胞渗透性autocamtide 2相关抑制肽II（Ant-AIP-II），未能类似地减少海马培养物中的钙电流或进入，如比率钙成像和全细胞膜片钳电生理学所示。值得注意的是，CK 59引起的抑制是可逆的;药物洗脱使钙水平在去极化后恢复至对照值。此外，CK 59的IC 50约为50 μM，仅为报告的CaMKII抑制IC 50值的5倍。其他CaMKII抑制剂KN 92和KN 62的类似非特异性作用先前已有报道。在所有三种激酶抑制剂的情况下，钙电流抑制的IC 50福尔斯接近CaMK II抑制的IC 50。我们的研究结果表明，CK 59减弱电压门控钙通道的活性，从而提供了更多的证据，以谨慎依赖于药理学抑制检查激酶依赖性现象时。
Investigation of kinase-related processes often uses pharmacological inhibition to reveal pathways in which kinases are involved. However, one concern about using such kinase inhibitors is their potential lack of specificity. Here, we report that the calcium-calmodulin dependent kinase II (CaMKII) inhibitor CK59 inhibited multiple voltage-gated calcium channels, including the L-type channel during depolarization in a dose-dependent manner. The use of another CaMKII inhibitor, cell permeable autocamtide 2-related inhibitory peptide II (Ant-AIP-II), failed to similarly decrease calcium current or entry in hippocampal cultures, as shown by ratiometric calcium imaging and whole cell patch clamp electrophysiology. Notably, inhibition due to CK59 was reversible; washout of the drug brought calcium levels back to control values upon depolarization. Furthermore, the IC50 for CK59 was approximately 50 μM, which is only five fold larger than the reported IC50 values for CaMKII inhibition. Similar nonspecific actions of other CaMKII inhibitors KN92 and KN62 have previously been reported. In the case of all three kinase inhibitors, the IC50 for calcium current inhibition falls near that of CaMKII inhibition. Our findings demonstrate that CK59 attenuates activity of voltage-gated calcium channels, and thus provide more evidence for caution when relying on pharmacological inhibition to examine kinase-dependent phenomena.
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影响因子：
3
作者：
Bray, Jennifer G.;Mynlieff, Michelle
通讯作者：
Mynlieff, Michelle
影响因子：
4.8
作者：
Konstantopoulos, Nicky;Marcuccio, Seb;Macaulay, S. Lance
通讯作者：
Macaulay, S. Lance
影响因子：
5.5
作者：
Park, Jin-Yong;Kang, Ho-Won;Lee, Jung-Ha
通讯作者：
Lee, Jung-Ha
影响因子：
4.7
作者：
SIHRA, TS;PEARSON, HA
通讯作者：
PEARSON, HA
影响因子：
3.3
作者：
Bray, J. G.;Mynlieff, M.
通讯作者：
Mynlieff, M.