• DOI:10.1126/SCIENCE.281.5374.269
• Corpus ID: 17912389

Specific covalent labeling of recombinant protein molecules inside live cells.

@article{Griffin1998SpecificCL,
  title={Specific covalent labeling of recombinant protein molecules inside live cells.},
  author={B A Griffin and Stephen R. Adams and Roger Tsien},
  journal={Science},
  year={1998},
  volume={281 5374},
  pages={
          269-72
        }
}

• B. A. Griffin, S. Adams, R. Tsien
• Published 10 July 1998
• Biology, Chemistry
• Science

Recombinant proteins containing four cysteines at the i, i + 1, i + 4, and i + 5 positions of an alpha helix were fluorescently labeled in living cells by extracellular administration of 4',5'-bis(1,3, 2-dithioarsolan-2-yl)fluorescein. This designed small ligand is membrane-permeant and nonfluorescent until it binds with high affinity and specificity to the tetracysteine domain. Such in situ labeling adds much less mass than does green fluorescent protein and offers greater versatility in attachment sites as well as potential spectroscopic and chemical properties. This system provides a recipe for slightly modifying a target protein so that it can be singled out from the many other proteins inside live cells and fluorescently stained by small nonfluorescent dye molecules added from outside the cells.