A photoactivatable GFP for selective photolabeling of proteins and cells
A photoactivatable GFP for selective photolabeling of proteins and cells
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DOI：
10.1126/science.1074952
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发表时间：
2002-09-13
期刊：
影响因子：
56.9
通讯作者：
Lippincott-Schwartz, J
中科院分区：
文献类型：
--
作者：
Patterson, GH;Lippincott-Schwartz, J
我们报道了Aequorea Victoria绿色荧光蛋白(GFP)的一种光激活变体，它在413纳米光的强烈照射下，在488纳米光激发下荧光增强100倍，并在有氧条件下保持数天稳定。这些特性为通过跟踪细胞中唯一可见的GFP的光活化分子来探索细胞内蛋白质动力学提供了新的工具。在这里，我们使用可光激活的GFP作为游离蛋白质来测量蛋白质在核膜上的扩散，并作为与溶酶体膜蛋白的嵌合体来展示快速的溶酶体膜交换。
We report a photoactivatable variant of the Aequorea victoria green fluorescent protein (GFP) that, after intense irradiation with 413-nanometer light, increases fluorescence 100 times when excited by 488-nanometer light and remains stable for days under aerobic conditions. These characteristics offer a new tool for exploring intracellular protein dynamics by tracking photoactivated molecules that are the only visible GFPs in the cell. Here, we use the photoactivatable GFP both as a free protein to measure protein diffusion across the nuclear envelope and as a chimera with a lysosomal membrane protein to demonstrate rapid interlysosomal membrane exchange.