Subcellular Protein Localization by Using a Genetically Encoded Fluorescent Amino Acid
2011

New Method for Labeling Proteins with Fluorescent Tags

publication Evidence: moderate

Author Information

Author(s): Charbon Godefroid, Brustad Eric, Scott Kevin A, Wang Jiangyun, Løbner-Olesen Anders, Schultz Peter G, Jacobs-Wagner Christine, Chapman Eli

Primary Institution: Yale University

Hypothesis

Can we label proteins in vivo without affecting their function using a genetically encoded fluorescent amino acid?

Conclusion

The study successfully demonstrates that a single unnatural fluorescent amino acid can be incorporated into a protein sequence without impairing its functionality.

Supporting Evidence

  • The incorporation of the fluorescent amino acid did not affect the protein's ability to function in vivo.
  • FtsZ10CouAA was visualized in E. coli, demonstrating the method's effectiveness.
  • The study showed that FtsZ10CouAA could substitute for wild-type FtsZ in E. coli.

Takeaway

Scientists found a way to add a tiny fluorescent tag to proteins so they can see where the proteins are in living cells without messing them up.

Methodology

The study used an orthogonal tRNA/aminoacyl-tRNA synthetase pair to incorporate a fluorescent amino acid into a protein sequence in E. coli.

Limitations

The efficiency of unnatural amino acid incorporation and the photophysical properties of the fluorophore may limit the technique's application.

Digital Object Identifier (DOI)

10.1002/cbic.201100282

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