cJun N-terminal kinase (JNK) phosphorylation of serine 36 is critical for p66Shc activation.
cJun N-terminal kinase (JNK) phosphorylation of serine 36 is critical for p66Shc activation.
复制标题
DOI：
10.1038/srep20930
复制
发表时间：
2016-02-12
影响因子：
4.6
通讯作者：
Troppmair J
中科院分区：
文献类型：
--
作者：
Khalid S;Drasche A;Thurner M;Hermann M;Ashraf MI;Fresser F;Baier G;Kremser L;Lindner H;Troppmair J
p66 Shc依赖性ROS的产生导致许多病理学，包括实体器官移植期间的缺血/再灌注损伤（IRI）。抑制p66 Shc活化可能提供一种新的治疗方法来预防损伤，这是体内抗氧化剂管理不善。以前的工作表明，促氧化和促凋亡功能的p66 Shc需要线粒体输入，这取决于丝氨酸36磷酸化。PKC β被认为是S36激酶，但cJun N-末端激酶（JNK）也可以磷酸化该残基。为了模拟缺血/再灌注（IR）的早期阶段，我们使用H2 O2处理或缺氧/复氧（HR）。在体内再灌注期间，我们观察到小鼠胚胎成纤维细胞（MEFs）和HL-1心肌细胞中JNK和p38活性增加沿着p66 ShcS 36磷酸化、ROS产生和细胞损伤显著增加。特异性抑制剂的应用仅在JNK 1/2的情况下引起p66 ShcS 36磷酸化的显著降低。此外，重组p66 Shc的S36磷酸化由JNK 1，而不是PKC β被证明。我们进一步证实了JNK 1/2依赖性调节p66 ShcS 36磷酸化，ROS的产生和细胞死亡使用JNK 1/2缺陷MEFs。最后，JNK 1/2敲除MEFs的低ROS表型被磷酸化模拟物p66 ShcS 36 E突变体逆转。因此，抑制JNK 1/2调节的p66 Shc活化可能提供一种预防氧化损伤的治疗方法。
p66Shc-dependent ROS production contributes to many pathologies including ischemia/reperfusion injury (IRI) during solid organ transplantation. Inhibiting p66Shc activation may provide a novel therapeutic approach to prevent damage, which is poorly managed by antioxidants in vivo. Previous work suggested that pro-oxidant and a pro-apoptotic function of p66Shc required mitochondrial import, which depended on serine 36 phosphorylation. PKCß has been proposed as S36 kinase but cJun N-terminal kinases (JNKs) may also phosphorylate this residue. To simulate the early stages of ischemia/reperfusion (IR) we either used H2O2 treatment or hypoxia/reoxygenation (HR). As during reperfusion in vivo, we observed increased JNK and p38 activity in mouse embryonic fibroblasts (MEFs) and HL-1 cardiomyocytes along with significantly increased p66ShcS36 phosphorylation, ROS production and cell damage. Application of specific inhibitors caused a pronounced decrease in p66ShcS36 phosphorylation only in the case of JNK1/2. Moreover, S36 phosphorylation of recombinant p66Shc by JNK1 but not PKCß was demonstrated. We further confirmed JNK1/2-dependent regulation of p66ShcS36 phosphorylation, ROS production and cell death using JNK1/2 deficient MEFs. Finally, the low ROS phenotype of JNK1/2 knockout MEFs was reversed by the phosphomimetic p66ShcS36E mutant. Inhibiting JNK1/2-regulated p66Shc activation may thus provide a therapeutic approach for the prevention of oxidative damage.