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Science 13 September 2002:
Vol. 297. no. 5588, pp. 1873 - 1877
DOI: 10.1126/science.1074952
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Reports

A Photoactivatable GFP for Selective Photolabeling of Proteins and Cells

George H. Patterson, Jennifer Lippincott-Schwartz*

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.

Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA.
*   To whom correspondence should be addressed. E-mail: jlippin{at}helix.nih.gov

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