title: Engineered metal binding sites on green fluorescence protein.
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.
authors.affiliation: Joint Science Department, Claremont McKenna, Pitzer, and Scripps Colleges, 925 North Mills Avenue, Claremont, California 91711, USA. richmond@jsd.claremont.edu
authors.forename: T A
authors.initials: TA
authors.lastname: Richmond
authors.forename: T T
authors.initials: TT
authors.lastname: Takahashi
authors.forename: R
authors.initials: R
authors.lastname: Shimkhada
authors.forename: J
authors.initials: J
authors.lastname: Bernsdorf
mesh_terms: D001665:Binding Sites; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism*; D008670:Metals / Q000378:metabolism*; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D015202:Protein Engineering / Q000379:methods
chemical_list: D008164:Luminescent Proteins; D008670:Metals; D049452:Green Fluorescent Proteins