Fluorescent tandem phycobiliprotein conjugates. Emission wavelength shifting by energy transfer.
A fluorescent tandem phycobiliprotein conjugate with a large Stokes shift was prepared by the covalent attachment of phycoerythrin to allophycocyanin. The efficiency of energy transfer from phycoerythrin to allophycocyanin in this disulfide-linked conjugate was 90%. A distinctive feature of this phycocyanin conjugate is the wide separation between the intense absorption maximum of phycoerythrin (epsilon = 2.4 x 10(6) cm-1 M-1 at 545 nm) and the fluorescence emission maximum of allophycocyanin (660 nm). Energy transfer from a donor to a covalently attached acceptor can be used to adjust the magnitude of the Stokes shift. Tandem phycobiliprotein conjugates can be used to advantage in fluorescence-activated cell sorting, fluorescence microscopy, and fluorescence immunoassay analyses.
D000458:Cyanobacteria / Q000378:metabolism; D004735:Energy Transfer; D045342:Light-Harvesting Protein Complexes; D010798:Phycocyanin / Q000378:metabolism; D010799:Phycoerythrin / Q000378:metabolism; D010860:Pigments, Biological / Q000378:metabolism*; D010940:Plant Proteins / Q000378:metabolism*; D013053:Spectrophotometry
D045342:Light-Harvesting Protein Complexes; D010860:Pigments, Biological; D010940:Plant Proteins; C002497:allophycocyanin; D010798:Phycocyanin; D010799:Phycoerythrin