Shen Chenxi, Li Zaiquan, Yang Xiaoda, Wang Kui
State Key Laboratories for Natural and Biomimetic Drugs and Department of Chemical Biology, School of Pharmaceutical Science, Peking University, Beijing 100191, PR China.
Chem Biol Interact. 2008 Nov 25;176(2-3):196-203. doi: 10.1016/j.cbi.2008.07.014. Epub 2008 Aug 19.
The effects of La3+ on the unfolded protein response signaling pathways were investigated in human hepatoblastoma HepG2 cells. Our data showed that La3+ could induce unfolded protein response in HepG2 cells, including a significant increase of BiP/GRP78 level, which is an important ER residential chaperone and an ER stress hallmark, in a concentration and time-dependent manner, UPR transducer IRE1 phosphorylation and splicing activation IRE1 downstream substrate XBP1 mRNA. By using La3+-affinity chromatography, the possible cellular target of La3+ leading to UPR events was shown to be the ER residential chaperone BiP/GRP78. BiP/GRP78 was shown to be a La3+ binding protein and the interaction of La3+ with BiP/GRP78 resulted in dissociation of BiP-IRE1 complexes. La3+ induced dissociation of the BiP/GRP78-IRE1 complex was in a time and concentration manner. The apparent dissociation constant was estimated to be 4 nM. In addition, La3+ was observed to slightly stimulate the production of cellular ROS and cause alteration of intracellular Ca2+, indicating the possible involvement of ROS and Ca2+ alteration in La3+ induced UPR. The present work provides a new perspective for understanding the biological and toxicological effects of La3+.
在人肝癌细胞HepG2中研究了La3+对未折叠蛋白反应信号通路的影响。我们的数据表明,La3+可在HepG2细胞中诱导未折叠蛋白反应,包括以浓度和时间依赖性方式显著增加BiP/GRP78水平,BiP/GRP78是一种重要的内质网驻留伴侣蛋白和内质网应激标志,同时诱导未折叠蛋白反应转导因子IRE1磷酸化并激活IRE1下游底物XBP1 mRNA的剪接。通过使用La3+亲和色谱法,导致未折叠蛋白反应事件的La3+可能的细胞靶点被证明是内质网驻留伴侣蛋白BiP/GRP78。BiP/GRP78被证明是一种La3+结合蛋白,La3+与BiP/GRP78的相互作用导致BiP-IRE1复合物解离。La3+诱导的BiP/GRP78-IRE1复合物解离呈时间和浓度依赖性。表观解离常数估计为4 nM。此外,观察到La3+轻微刺激细胞活性氧的产生并导致细胞内Ca2+改变,表明活性氧和Ca2+改变可能参与La3+诱导的未折叠蛋白反应。本研究为理解La3+的生物学和毒理学效应提供了新的视角。