Specific Covalent Labeling of Recombinant Protein Molecules Inside Live Cells

doi:10.1126/science.281.5374.269

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Science 10 July 1998:
Vol. 281. no. 5374, pp. 269 - 272
DOI: 10.1126/science.281.5374.269

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Reports

Specific Covalent Labeling of Recombinant Protein Molecules Inside Live Cells

B. Albert Griffin, ^* Stephen R. Adams, Roger Y. Tsien

Recombinant proteins containing four cysteines at the i, i + 1, i + 4, and i + 5 positions of an $alpha$ helix were fluorescentlylabeled 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 nonfluorescentuntil it binds with high affinity and specificity to the tetracysteinedomain. Such in situ labeling adds much less mass than does greenfluorescent protein and offers greater versatility in attachmentsites as well as potential spectroscopic and chemical properties.This system provides a recipe for slightly modifying a targetprotein so that it can be singled out from the many other proteinsinside live cells and fluorescently stained by small nonfluorescentdye molecules added from outside the cells.

B. A. Griffin, Department of Chemistry and Biochemistry, University of California San Diego, La Jolla, CA 92093-0647, USA. S. R. Adams, Department of Pharmacology, University of California San Diego, La Jolla, CA 92093-0647, USA. R. Y. Tsien, Department of Pharmacology, Department of Chemistry and Biochemistry, and Howard Hughes Medical Institute, University of California San Diego, La Jolla, CA 92093-0647, USA.
^* Present address: Aurora Biosciences, 11010 Torreyana Road, San Diego, CA 92121, USA.

To whom correspondence should be addressed.