Shi Xiaoyu, Sun Yue, Wang Ping, Gu Lingling, Wang Lu, Yang Huan, Wei Qun, Li Zhimei, Luo Jing
Gene Engineering and Biotechnology, Beijing Key Laboratory, College of Life Sciences, Beijing Normal University, No.19, Xinjiekouwaidajie, Beijing 100875, China; College of Life Sciences, Langfang Normal University, Hebei, 065000, China.
Gene Engineering and Biotechnology, Beijing Key Laboratory, College of Life Sciences, Beijing Normal University, No.19, Xinjiekouwaidajie, Beijing 100875, China.
Biochem Biophys Res Commun. 2018 Feb 19;496(4):1109-1114. doi: 10.1016/j.bbrc.2018.01.148. Epub 2018 Feb 7.
Calcineurin (CN) is a protein phosphatase and widely distributed in eukaryotes, with an extremely high level of expression in mammalian brain. Alpha-synuclein (α-syn) is a small soluble protein expressed primarily at presynaptic terminals in the central nervous system. In our present study, we explored the interactions between CN and α-syn in vitro. Based on the data from microscale thermophoresis, GST pull-down assays, and co-immunoprecipitation, we found that CN binds α-syn. Furthermore, this interaction is mediated by calcium/calmodulin (Ca/CaM) signaling. Additionally, thapsigargin (TG) triggered an increase in CN activity and α-syn aggregation in HEK293 cells stably transfected with α-syn. Our previous study in vivo suggest that overexpression of α-syn in transgenic mice significantly promoted CN activity and subsequent nuclear translocation of nuclear factor of activated T-cells (NFAT) in the midbrain dopaminergic (mDA) neurons. These in vivo and in vitro studies have been complementary with each other, representing the changes in the CN-dependent pathway affected by overexpression of α-syn.
钙调神经磷酸酶（CN）是一种蛋白磷酸酶，广泛分布于真核生物中，在哺乳动物大脑中表达水平极高。α-突触核蛋白（α-syn）是一种主要在中枢神经系统突触前终末表达的小的可溶性蛋白。在我们目前的研究中，我们在体外探索了CN与α-syn之间的相互作用。基于微量热泳动、GST下拉实验和免疫共沉淀的数据，我们发现CN与α-syn结合。此外，这种相互作用是由钙/钙调蛋白（Ca/CaM）信号介导的。另外，毒胡萝卜素（TG）在稳定转染α-syn的HEK293细胞中引发了CN活性增加和α-syn聚集。我们之前的体内研究表明，转基因小鼠中α-syn的过表达显著促进了中脑多巴胺能（mDA）神经元中CN活性以及随后活化T细胞核因子（NFAT）的核转位。这些体内和体外研究相互补充，代表了受α-syn过表达影响的CN依赖性途径的变化。
