Lu Jingze, He Zixuan, Fan Junmei, Xu Pingyong, Chen Liangyi
National Key Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Science, Datun Road 15#, Beijing 100101, PR China.
Biochem Biophys Res Commun. 2008 Jun 27;371(2):315-9. doi: 10.1016/j.bbrc.2008.04.077. Epub 2008 Apr 28.
Previously, we identified a clathrin-dependent slow endocytosis and a clathrin-independent fast endocytosis in pancreatic beta cells, both triggered by elevated cytoplasmic Ca(2+) concentration. In the current study, we attempted to explore the roles of different dynamin isoforms in these endocytotic processes. We first confirmed the existence of both neuron-specific dynamin 1 and ubiquitous dynamin 2 in INS-1 cells using both quantitative RT-PCR and Western blot experiments. By specifically knocking down the endogenous level of either dynamin isoform from INS-1 cells, we showed that dynamin 1 and dynamin 2 simultaneously participate in the clathrin-independent and -dependent membrane retrieval in pancreatic beta cells. Transferrin internalization was also inhibited in cells with knock down of both dynamin 1 and dynamin 2. Based on these results, we argue that different dynamin isoforms play overlapping roles in different types of endocytosis.
此前,我们在胰腺β细胞中鉴定出一种网格蛋白依赖性的慢内吞作用和一种网格蛋白非依赖性的快内吞作用,二者均由细胞质中Ca(2+)浓度升高触发。在当前研究中,我们试图探究不同动力蛋白亚型在这些内吞过程中的作用。我们首先通过定量RT-PCR和蛋白质印迹实验证实了INS-1细胞中存在神经元特异性动力蛋白1和普遍存在的动力蛋白2。通过特异性敲低INS-1细胞中任一动力蛋白亚型的内源性水平,我们发现动力蛋白1和动力蛋白2同时参与胰腺β细胞中网格蛋白非依赖性和依赖性的膜回收过程。在同时敲低动力蛋白1和动力蛋白2的细胞中,转铁蛋白的内化也受到抑制。基于这些结果,我们认为不同的动力蛋白亚型在不同类型的内吞作用中发挥重叠作用。