PKC modulation of calcium current and anesthetic action

PKC 调节钙电流和麻醉作用

• 批准号: 7266199
• 负责人: GANESAN Lenin KAMATCHI
• 金额: $ 20.45万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-08-01 至 2008-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7266199
• 关键词: PKC; modulation; calcium; current; anesthetic

DESCRIPTION (provided by applicant): Voltage-gated Ca2+-channels (Cav) and protein kinase C (PKC) can both be regulated by volatile anesthetics, but the nature of the effect has been controversial. We hypothesize that anesthetic inhibition of Cav channels and synaptic transmission are mediated by specific PKC isozymes. Although potential PKC phosphorylation sites are present in the pore-forming alpha1 and the auxiliary beta and alpha2/delta subunits of Cav channels; alpha1subunits seem to play the major role in PKC-induced regulation of these channels. In Aim I of this proposal, we will identify phosphorylation sites in the a1 subunits of Cav channels by direct (with phorbol 12-myristate 13-acetate; PMA) or receptor-stimulated (with muscarinic M1) activation of PKC in Xenopus oocytes coexpressing these receptors and Cav channels. Possible PKC phosphorylation sites will be mutated to Ala or Glu and whole-cell Ba2+ (a substitute for Ca2+)-current in response to two types of PKC activation will be investigated using voltage-clamp measurements. In Aim II we will identify PKC isozymes involved in the direct or receptor-induced modulation of PKC-sensitive Cav channels expressed in Xenopus oocytes. This will be examined using experiments involving down-regulation of PKC, activation-induced translocation, selective loss/inhibition of individual PKC isozymes and 'add-back' studies. In Aim III we will identify the PKC isozymes modulated by volatile anesthetics, halothane or isoflurane in Xenopus oocytes by employing the methods mentioned in Aim II. Based on the results of our studies, isozyme selective activators or inhibitors of PKC may be used along with the volatile general anesthetics to i) potentiate the action of anesthetics and ii) to reduce the side effects of these agents caused by their non-specific actions.

描述（由申请人提供）： 电压门控性钙通道（Cav）和蛋白激酶C（PKC）都可以被挥发性麻醉药调节，但其作用的性质一直存在争议。我们假设麻醉剂对Cav通道和突触传递的抑制是由特定的PKC同工酶介导的。虽然潜在的PKC磷酸化位点存在于Cav通道的成孔α 1和辅助β和α 2/δ亚基中，但α 1亚基似乎在PKC诱导的这些通道调节中起主要作用。在本提案的目的I中，我们将通过直接（用佛波醇12-肉豆蔻酸酯13-乙酸酯; PMA）或受体刺激（用毒蕈碱M1）激活共表达这些受体和Cav通道的爪蟾卵母细胞中的PKC来鉴定Cav通道a1亚基中的磷酸化位点。可能的PKC磷酸化位点将突变为Ala或Glu，并且将使用电压钳测量来研究响应两种类型PKC激活的全细胞Ba 2+（Ca 2+的替代品）电流。在目的II中，我们将确定参与直接或受体诱导的PKC敏感的Cav通道在非洲爪蟾卵母细胞中表达的调制PKC同工酶。这将通过涉及PKC下调、激活诱导的易位、单个PKC同工酶的选择性丢失/抑制和“加回”研究的实验进行检查。在目的III中，我们将确定PKC同工酶的挥发性麻醉剂，氟烷或异氟烷调节爪蟾卵母细胞采用的方法在目的II。基于我们的研究结果，PKC的同工酶选择性激活剂或抑制剂可以与挥发性全身麻醉剂一起沿着使用，以i）增强麻醉剂的作用和ii）减少这些药剂由其非特异性作用引起的副作用。

项目成果

Identification of sites responsible for potentiation of type 2.3 calcium currents by acetyl-beta-methylcholine.

鉴定负责乙酰-β-甲基胆碱增强 2.3 型钙电流的位点。

• DOI: 10.1074/jbc.m308606200
• 发表时间: 2004
• 期刊: The Journal of biological chemistry
• 作者: Kamatchi,GanesanL;Franke,Ruthie;Lynch3rd,Carl;Sando,JulianneJ
• 通讯作者: Sando,JulianneJ

Effects of isoflurane on the expressed Cav2.2 currents in Xenopus oocytes depend on the activation of protein kinase Cδ and its phosphorylation sites in the Cav2.2α1 subunits.

异氟烷对非洲爪蟾卵母细胞中表达的 Cav2.2 电流的影响取决于蛋白激酶 Cδ 的激活及其在 Cav2.2α1 亚基中的磷酸化位点。

• DOI: 10.1016/j.neuroscience.2011.02.041
• 发表时间: 2011
• 期刊: Neuroscience
• 影响因子: 3.3
• 作者: Rajagopal,S;Fang,H;Lynch3rd,C;Sando,JJ;Kamatchi,GL
• 通讯作者: Kamatchi,GL

Inhibitory role of Ser-425 of the alpha1 2.2 subunit in the enhancement of Cav 2.2 currents by phorbol-12-myristate, 13-acetate.

α1 2.2 亚基的 Ser-425 在佛波醇 12-肉豆蔻酸酯、13-乙酸酯增强 Cav 2.2 电流中的抑制作用。

• DOI: 10.1074/jbc.m601776200
• 发表时间: 2006
• 期刊: The Journal of biological chemistry
• 作者: Fang,Hongyu;Patanavanich,Saharat;Rajagopal,Senthilkumar;Yi,Xiaobin;Gill,MonicaS;Sando,JulianneJ;Kamatchi,GanesanL
• 通讯作者: Kamatchi,GanesanL

Protein kinase C isozyme-specific potentiation of expressed Ca v 2.3 currents by acetyl-beta-methylcholine and phorbol-12-myristate, 13-acetate.

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
• 发表时间: 2008
• 期刊: Brain research
• 影响因子: 2.9
• 作者: Rajagopal,Senthilkumar;Fang,Hongyu;Patanavanich,Saharat;Sando,JulianneJ;Kamatchi,GanesanL
• 通讯作者: Kamatchi,GanesanL

Site-specific regulation of CA(V)2.2 channels by protein kinase C isozymes betaII and epsilon.

蛋白激酶 C 同工酶 betaII 和 epsilon 对 CA(V)2.2 通道的位点特异性调节。

• DOI: 10.1016/j.neuroscience.2008.12.047
• 发表时间: 2009
• 期刊: Neuroscience
• 影响因子: 3.3
• 作者: Rajagopal,S;Fang,H;Oronce,CIA;Jhaveri,S;Taneja,S;Dehlin,EM;Snyder,SL;Sando,JJ;Kamatchi,GL
• 通讯作者: Kamatchi,GL