Moon Jae Yun, Choi Su Jin, Heo Cheol Ho, Kim Hwan Myung, Kim Hye Sun
Department of Biological Science, College of Natural Sciences, Ajou University, Suwon, Korea.
Departments of Chemistry and Energy Systems Research, Ajou University, Suwon, Korea.
FEBS J. 2017 Jul;284(13):2052-2065. doi: 10.1111/febs.14103. Epub 2017 Jun 4.
α-Syntrophin is a component of the dystrophin-glycoprotein complex that interacts with various intracellular signaling proteins in muscle cells. The α-syntrophin knock-down C2 cell line (SNKD), established by infecting lentivirus particles with α-syntrophin shRNA, is characterized by a defect in terminal differentiation and increase in cell death. Since myoblast differentiation is accompanied by intensive mitochondrial biogenesis, the generation of intracellular reactive oxygen species (ROS) is also increased during myogenesis. Two-photon microscopy imaging showed that excessive intracellular ROS accumulated during the differentiation of SNKD cells as compared with control cells. The formation of 4-hydroxynonenal adduct, a byproduct of lipid peroxidation during oxidative stress, significantly increased in differentiated SNKD myotubes and was dramatically reduced by epigallocatechin-3-gallate, a well-known ROS scavenger. Among antioxidant enzymes, catalase was significantly decreased during differentiation of SNKD cells without changes at the mRNA level. Of interest was the finding that the degradation of catalase was rescued by MG132, a proteasome inhibitor, in the SNKD cells. This study demonstrates a novel function of α-syntrophin. This protein plays an important role in the regulation of oxidative stress from endogenously generated ROS during myoblast differentiation by modulating the protein stability of catalase.
α-肌养蛋白是肌营养不良蛋白-糖蛋白复合物的一个组成部分,它在肌肉细胞中与多种细胞内信号蛋白相互作用。通过用α-肌养蛋白短发夹RNA感染慢病毒颗粒建立的α-肌养蛋白敲低C2细胞系(SNKD),其特征在于终末分化缺陷和细胞死亡增加。由于成肌细胞分化伴随着强烈的线粒体生物发生,在肌生成过程中细胞内活性氧(ROS)的产生也会增加。双光子显微镜成像显示,与对照细胞相比,SNKD细胞分化过程中细胞内ROS过度积累。4-羟基壬烯醛加合物是氧化应激期间脂质过氧化的副产物,在分化的SNKD肌管中显著增加,并被著名的ROS清除剂表没食子儿茶素-3-没食子酸酯显著降低。在抗氧化酶中,过氧化氢酶在SNKD细胞分化过程中显著降低,而mRNA水平没有变化。有趣的是,蛋白酶体抑制剂MG132在SNKD细胞中挽救了过氧化氢酶的降解。这项研究证明了α-肌养蛋白的一种新功能。这种蛋白质在成肌细胞分化过程中,通过调节过氧化氢酶的蛋白质稳定性,对内源性产生的ROS引起的氧化应激调节中起重要作用。