Bichromophoric dyes for wavelength shifting of dye-protein fluoromodules

Dye-protein fluoromodules consist of fluorogenic dyes and single chain antibody fragments that form brightly fluorescent noncovalent complexes. This report describes two new bichromophoric dyes that extend the range of wavelengths of excitation or emission of existing fluoromodules. In one case, a f...

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Bibliographic Details
Published in:	Organic & biomolecular chemistry Vol. 13; no. 12; p. 3699
Main Authors:	Pham, Ha H, Szent-Gyorgyi, Christopher, Brotherton, Wendy L, Schmidt, Brigitte F, Zanotti, Kimberly J, Waggoner, Alan S, Armitage, Bruce A
Format:	Journal Article
Language:	English
Published:	England 28-03-2015
Subjects:	Benzothiazoles - chemistry Coumarins - chemistry Fluorescence Resonance Energy Transfer Fluorescent Dyes - chemical synthesis Fluorescent Dyes - chemistry Light Microscopy, Confocal Proteins - metabolism Quinolines - chemistry Saccharomyces cerevisiae - cytology Spectrophotometry, Ultraviolet
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Summary:	Dye-protein fluoromodules consist of fluorogenic dyes and single chain antibody fragments that form brightly fluorescent noncovalent complexes. This report describes two new bichromophoric dyes that extend the range of wavelengths of excitation or emission of existing fluoromodules. In one case, a fluorogenic thiazole orange (TO) was attached to an energy acceptor dye, Cy5. Upon binding to a protein that recognizes TO, red emission due to efficient energy transfer from TO to Cy5 replaces the green emission observed for monochromophoric TO bound to the same protein. Separately, TO was attached to a coumarin that serves as an energy donor. The same green emission is observed for coumarin-TO and TO bound to a protein, but efficient energy transfer allows violet excitation of coumarin-TO, versus longer wavelength, blue excitation of monochromophoric TO. Both bichromophores exhibit low nanomolar KD values for their respective proteins, >95% energy transfer efficiency and high fluorescence quantum yields.
ISSN:	1477-0539
DOI:	10.1039/c4ob02522a