Mitochondrial import of PKCepsilon is mediated by HSP90: a role in cardioprotection from ischaemia and reperfusion injury.
Mitochondrial import of PKCepsilon is mediated by HSP90: a role in cardioprotection from ischaemia and reperfusion injury.
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DOI：
10.1093/cvr/cvq154
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发表时间：
2010-10-01
影响因子：
10.8
通讯作者：
Mochly-Rosen D
中科院分区：
文献类型：
--
作者：
Budas GR;Churchill EN;Disatnik MH;Sun L;Mochly-Rosen D
蛋白激酶Cε (PKCε)对心肌缺血再灌注（IR）损伤的保护至关重要。介导细胞保护的PKCε底物位于线粒体中。然而，使线粒体易位和PKCε的输入使这些底物磷酸化的机制尚不清楚。热休克蛋白90 （HSP90）是一种细胞保护蛋白伴侣，参与线粒体许多蛋白质的输入。在这里，我们研究了HSP90在PKCε线粒体输入中的作用。通过体外灌注IR大鼠心脏模型，我们发现pkce ε在IR后从细胞质转移到线粒体部分。免疫金电镜和线粒体分离表明，IR后，线粒体PKCε定位在线粒体内，在线粒体内膜上。HSP90的药理抑制阻止了ir诱导的PKCε与外膜转位酶（Tom20）之间的相互作用，减少了PKCε的线粒体输入，并使坏死细胞死亡增加了约70%。利用合理的方法，我们设计了一个7个氨基酸的PKCε肽激活剂，该激活剂来源于PKCε C2结构域的HSP90同源序列（称为εHSP90）。在离体和体内心肌梗死模型中，该肽（与细胞渗透TAT蛋白衍生肽TAT47-57结合）增加PKCε - hsp90蛋白-蛋白相互作用，增强PKCε的线粒体易位，增加线粒体内PKCε底物醛脱氢酶2的磷酸化和活性，减少心脏损伤。我们的研究结果表明，hsp90介导的PKCε的线粒体输入在心肌免受IR损伤的保护中起重要作用。
Protein kinase C epsilon (PKCε) is critical for cardiac protection from ischaemia and reperfusion (IR) injury. PKCε substrates that mediate cytoprotection reside in the mitochondria. However, the mechanism enabling mitochondrial translocation and import of PKCε to enable phosphorylation of these substrates is not known. Heat shock protein 90 (HSP90) is a cytoprotective protein chaperone that participates in mitochondrial import of a number of proteins. Here, we investigated the role of HSP90 in mitochondrial import of PKCε. Using an ex vivo perfused rat heart model of IR, we found that PKCε translocates from the cytosol to the mitochondrial fraction following IR. Immunogold electron microscopy and mitochondrial fractionation demonstrated that following IR, mitochondrial PKCε is localized within the mitochondria, on the inner mitochondrial membrane. Pharmacological inhibition of HSP90 prevented IR-induced interaction between PKCε and the translocase of the outer membrane (Tom20), reduced mitochondrial import of PKCε, and increased necrotic cell death by ∼70%. Using a rational approach, we designed a 7-amino acid peptide activator of PKCε, derived from an HSP90 homologous sequence located in the C2 domain of PKCε (termed ψεHSP90). Treatment with this peptide (conjugated to the cell permeating TAT protein-derived peptide, TAT47–57) increased PKCε–HSP90 protein–protein interaction, enhanced mitochondrial translocation of PKCε, increased phosphorylation and activity of an intra-mitochondrial PKCε substrate, aldehyde dehydrogenase 2, and reduced cardiac injury in ex vivo and in vivo models of myocardial infarction. Our results suggest that HSP90-mediated mitochondrial import of PKCε plays an important role in the protection of the myocardium from IR injury.
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影响因子：
4.8
作者：
Kheifets, Viktoria;Bright, Rachel;Mochly-Rosen, Daria
通讯作者：
Mochly-Rosen, Daria
DOI：
10.1073/pnas.191369098
发表时间：
2001-09-25
影响因子：
11.1
作者：
Chen, L;Hahn, H;Mochly-Rosen, D
通讯作者：
Mochly-Rosen, D
影响因子：
37.8
作者：
Inagaki, K;Begley, R;Mochly-Rosen, D
通讯作者：
Mochly-Rosen, D
影响因子：
3.3
作者：
Bhangoo, Melanie K.;Tzankov, Stefan;Young, Jason C.
通讯作者：
Young, Jason C.
影响因子：
5
作者：
Churchill, Eric N.;Disatnik, Marie-Helene;Mochly-Rosen, Daria
通讯作者：
Mochly-Rosen, Daria