A Photoactivatable GFP for Selective Photolabeling of Proteins and Cells

作者：George H. Patterson, Jennifer Lippincott‐Schwartz · 发表于：Science · 年份：2002 · DOI：10.1126/science.1074952 · 被引用次数：1616 · 研究领域：Advanced Fluorescence Microscopy Techniques、Photosynthetic Processes and Mechanisms、bioluminescence and chemiluminescence research

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.