A light-triggered protein secretion system.
A light-triggered protein secretion system.
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DOI：
10.1083/jcb.201210119
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发表时间：
2013-05-13
期刊：
影响因子：
--
通讯作者：
Kennedy MJ
中科院分区：
文献类型：
--
作者：
Chen D;Gibson ES;Kennedy MJ
与植物光感受器蛋白UVR 8融合的分泌蛋白在ER中被有条件地隔离，直到光脉冲触发通过分泌途径的运输，允许精确控制正向分泌运输。蛋白质相互作用的光学控制已经成为操纵和研究各种细胞过程的强大实验范例。现在已经有工具可以控制许多细胞功能，但一些基本的过程，如蛋白质分泌，很难使用当前的光学工具进行工程设计。在这里，我们使用UVR8，一种形成光不稳定同源二聚体的植物感光蛋白，来设计第一个光触发的蛋白质分泌系统。UVR8融合蛋白被有条件地隔离在内质网中，并且短暂的光脉冲触发了通过分泌途径到质膜的稳健的向前运输。UVR 8对用于成像青色、绿色或红色荧光蛋白变体的激发光没有响应，从而允许当其穿过细胞分泌途径时细胞标记物和分泌的蛋白质货物的可视化。我们在神经元中实施了这种新工具，以证明树突分支点附近的分泌性货物的受限的局部运输。
Secreted proteins fused to the plant photoreceptor protein UVR8 are conditionally sequestered in the ER until a pulse of light triggers trafficking through the secretory pathway, allowing precise control of forward secretory trafficking. Optical control of protein interactions has emerged as a powerful experimental paradigm for manipulating and studying various cellular processes. Tools are now available for controlling a number of cellular functions, but some fundamental processes, such as protein secretion, have been difficult to engineer using current optical tools. Here we use UVR8, a plant photoreceptor protein that forms photolabile homodimers, to engineer the first light-triggered protein secretion system. UVR8 fusion proteins were conditionally sequestered in the endoplasmic reticulum, and a brief pulse of light triggered robust forward trafficking through the secretory pathway to the plasma membrane. UVR8 was not responsive to excitation light used to image cyan, green, or red fluorescent protein variants, allowing multicolor visualization of cellular markers and secreted protein cargo as it traverses the cellular secretory pathway. We implemented this novel tool in neurons to demonstrate restricted, local trafficking of secretory cargo near dendritic branch points.
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影响因子：
48
作者：
Kennedy, Matthew J.;Hughes, Robert M.;Peteya, Leslie A.;Schwartz, Joel W.;Ehlers, Michael D.;Tucker, Chandra L.
通讯作者：
Tucker, Chandra L.
影响因子：
56.9
作者：
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通讯作者：
Clackson, T
影响因子：
56.9
作者：
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通讯作者：
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影响因子：
16.2
作者：
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通讯作者：
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影响因子：
64.8
作者：
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通讯作者：
Quail, PH