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