Real-time bioluminescence imaging of a protein secretory pathway in living mammalian cells using Gaussia luciferase
Real-time bioluminescence imaging of a protein secretory pathway in living mammalian cells using Gaussia luciferase
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DOI：
10.1016/j.febslet.2007.08.036
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发表时间：
2007-10-02
期刊：
影响因子：
3.5
通讯作者：
Inouye, Satoshi
中科院分区：
文献类型：
--
作者：
Suzuki, Takahiro;Usuda, Shinsuke;Inouye, Satoshi
使用光子计数和电荷耦合器件（CCD）相机，我们已经应用了实时生物发光成像的方法来研究哺乳动物细胞中的蛋白质运输。在中国仓鼠卵巢和PC12D细胞的活细胞中，使用分泌的Gaussia荧光素酶（GLase）作为报告蛋白，在一分钟内观察蛋白质的胞吐分泌和靶向细胞表面的蛋白质。在将细胞与蓝蛋白（腔肠素）孵育10分钟后，蓝蛋白被导入细胞中，并且细胞中的囊泡运输网络可以通过GLase活性的发光图像显示。此外，我们证明了具有异源信号肽序列的GLase靶向于神经元分化的PC12D细胞中的细胞表面，并且可以在几秒钟内检测到发光信号。(c)2007年欧洲生物化学学会联合会。Elsevier B.V.出版，保留所有权利。
Using photon counting and charge-coupled device (CCD) cameras, we have applied the method of real-time bioluminescence imaging to investigate protein trafficking in mammalian cells. In the living cells of Chinese hamster ovary and PC12D cells, exocytotic secretion of protein and protein targeting on the cell surface were visualized using the secreted Gaussia luciferase (GLase) as a reporter protein in a minute. After incubation of the cells with luciferin (coelenterazine) for 10 min, luciferin was imported into the cells and the vesicle transport network in the cells could be shown by luminescence images of GLase activity. Further, we demonstrate that GLase with a heterologous signal peptide sequence is targeted to the cell surface in neuronally differentiated PC12D cells and luminescence signals could be detected in a few seconds. (c) 2007 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.