Video-rate bioluminescence imaging of protein secretion from a living cell.
Video-rate bioluminescence imaging of protein secretion from a living cell.
复制标题
活细胞蛋白质分泌的视频生物发光成像。
DOI：
10.1007/978-1-62703-718-1_6
复制
发表时间：
2014
影响因子：
--
通讯作者：
S. Inouye
中科院分区：
文献类型：
--
作者：
Takahiro Suzuki;S. Inouye
我们已经建立了一种视频速率生物发光成像的方法，可视化从一个单一的活细胞使用高斯荧光素酶（GLase）作为报告蛋白的胞吐蛋白分泌。通过分泌的GLase（荧光素酶）和腔肠素（coelentrazin）的酶促反应产生的发光信号用电子倍增电荷耦合器件照相机检测。可以使用融合到GLase的蛋白质以30-500 ms的时间分辨率可视化蛋白质分泌的胞吐事件。生物发光视频图像的信号分析揭示了胞吐位点的数量、胞吐事件的频率和整个细胞上分泌的蛋白质的量。此外，该方法可以区分分泌的和细胞表面结合的蛋白质。我们的方法是研究活细胞整个表面蛋白质分泌和定位的直接方法。
We have established a method of video-rate bioluminescence imaging to visualize exocytotic protein secretion from a single living cell using Gaussia luciferase (GLase) as a reporter protein. The luminescence signals produced by the enzymatic reaction of secreted GLase (luciferase) and coelenterazine (luciferin) are detected with an electron-multiplying charge-coupled device camera. An exocytotic event of protein secretion can be visualized using the protein fused to GLase with a time resolution of 30-500 ms. Signal analyses of the bioluminescence video images reveal a number of exocytotic sites, the frequency of exocytotic events, and the amount of secreted protein on a whole cell. Furthermore, the method can distinguish between secreted and cell surface-bound proteins. Our method is a direct approach to investigate the secretion and localization of proteins on the whole surface of living cells.
影响因子：
12.4
作者：
Tannous, BA;Kim, DE;Breakefield, XO
通讯作者：
Breakefield, XO