DOI: 10.1073/pnas.0403668101. EosFP, a fluorescent marker protein with UV-inducible green-to-red fluorescence conversion

EosFP, a fluorescent marker protein with UV-inducible green-to-red fluorescence conversion

10.1073/pnas.0403668101

Crossref journal-article

Proceedings of the National Academy of Sciences

Proceedings of the National Academy of Sciences (341)

Abstract

A gene encoding a fluorescent protein from the stony coral Lobophyllia hemprichii has been cloned in Escherichia coli and characterized by biochemical and biophysical methods. The protein, which we named EosFP, emits strong green fluorescence (516 nm) that changes to red (581 nm) upon near-UV irradiation at ≈390 nm because of a photo-induced modification involving a break in the peptide backbone next to the chromophore. Single-molecule fluorescence spectroscopy shows that the wild type of EosFP is tetrameric, with strong Förster resonance coupling among the individual fluorophores. We succeeded in breaking up the tetramer into AB and AC subunit dimers by introducing the single point mutations V123T and T158H, respectively, and the combination of both mutations yielded functional monomers. Fusion constructs with a variety of proteins were prepared and expressed in human cells, showing that normal biological functions were retained. The possibility to locally change the emission wavelength by focused UV light makes EosFP a superb marker for experiments aimed at tracking the movements of biomolecules within the living cell.

Bibliography

Wiedenmann, J., Ivanchenko, S., Oswald, F., Schmitt, F., RÃ¶cker, C., Salih, A., Spindler, K.-D., & Nienhaus, G. U. (2004). EosFP, a fluorescent marker protein with UV-inducible green-to-red fluorescence conversion. Proceedings of the National Academy of Sciences, 101(45), 15905â15910.

Authors 8

Jörg Wiedenmann (first)
Sergey Ivanchenko (additional)
Franz Oswald (additional)
Florian Schmitt (additional)
Carlheinz Röcker (additional)
Anya Salih (additional)
Klaus-Dieter Spindler (additional)
G. Ulrich Nienhaus (additional)

References 32 Referenced 598

10.1016/0378-1119(92)90691-H
10.1126/science.8303295
10.1146/annurev.biochem.67.1.509
Wiedenmann, J., Röcker, C. & Funke, W. (1999) in Verhandlungen der Gesellschaft für Ökologie, ed. Pfadenhauer, J. (Springer, Heidelberg), Vol. 29, pp. 497–503. / Verhandlungen der Gesellschaft für Ökologie (1999)
Wiedenmann J. (1997) in Offenlegungsschrift DE 197 18 640 A1 (German Patent and Trade Mark Office Munich) pp. 1–18.
10.1038/13657
10.1016/S0014-5793(00)01895-0
10.1073/pnas.97.26.14091
10.1073/pnas.182157199
Wiedenmann, J., Ivanchenko, S., Oswald, F. & Nienhaus, G. U. (2004) Mar. Biotechnol. 6, 270–277.15136917 / Mar. Biotechnol. (2004)
10.1093/molbev/msh079
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10.1093/emboj/cdf549

Dates

Type	When
Created	20 years, 10 months ago (Oct. 25, 2004, 8:16 p.m.)
Deposited	3 years, 4 months ago (April 12, 2022, 9:25 a.m.)
Indexed	1 month, 1 week ago (July 22, 2025, 6:51 a.m.)
Issued	20 years, 10 months ago (Oct. 25, 2004)
Published	20 years, 10 months ago (Oct. 25, 2004)
Published Online	20 years, 10 months ago (Oct. 25, 2004)
Published Print	20 years, 9 months ago (Nov. 9, 2004)

Funders 0

None

BibTeX

@article{Wiedenmann_2004, title={EosFP, a fluorescent marker protein with UV-inducible green-to-red fluorescence conversion}, volume={101}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.0403668101}, DOI={10.1073/pnas.0403668101}, number={45}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Wiedenmann, Jörg and Ivanchenko, Sergey and Oswald, Franz and Schmitt, Florian and Röcker, Carlheinz and Salih, Anya and Spindler, Klaus-Dieter and Nienhaus, G. Ulrich}, year={2004}, month=oct, pages={15905–15910} }

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