Engineered metal binding sites on green fluorescence protein

Biochem Biophys Res Commun. 2000 Feb 16;268(2):462-5. doi: 10.1006/bbrc.1999.1244.

Abstract

The ability to assay a variety of metals by noninvasive methods has applications in both biomedical and environmental research. Green fluorescent protein (GFP) is a protein isolated from coelenterates that exhibits spontaneous fluorescence. GFP does not require any exogenous cofactors for fluorescence, and can be easily appended to other proteins at the DNA level, producing a fluorescence-labeled target protein in vivo. Metals in close proximity to chromophores are known to quench fluorescence in a distance-dependent fashion. Potential metal binding sites on the surface of GFP have been identified and mutant proteins have been designed, created, and characterized. These metal-binding mutants of GFP exhibit fluorescence quenching at lower transition metal ion concentrations than those of the wild-type protein. These GFP mutants represent a new class of protein-based metal sensors.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Binding Sites
  • Green Fluorescent Proteins
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism*
  • Metals / metabolism*
  • Models, Molecular
  • Mutagenesis, Site-Directed
  • Protein Engineering / methods

Substances

  • Luminescent Proteins
  • Metals
  • Green Fluorescent Proteins