Nienhaus, G. Ulrich, Nienhaus, Karin, Hölzle, Angela, Ivanchenko, Sergey, Renzi, Fabiana, Oswald, Franz, Wolff, Michael, Schmitt, Florian, Röcker, Carlheinz, Vallone, Beatrice, Weidemann, Wolfgang, Heilker, Ralf, Nar, Herbert and Wiedenmann, Jörg
Photoconvertible fluorescent protein EosFP: biophysical properties and cell biology applications
Photochemistry and Photobiology, 82, (2), . (doi:10.1562/2005-05-19-RA-533).
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EosFP is a fluorescent protein from the coral Lobophyllia hemprichii that changes its fluorescence emission from green to red upon irradiation with near-UV light. Here we present the spectroscopic properties of wild-type EosFP and a variety of monomeric and dimeric mutants and provide a structural interpretation of its oligomerization and photoconversion, which is based on X-ray structure analysis of the green and red species that we reported recently. Because functional expression of the monomeric EosFP variant is limited to temperatures of 30°C, we have developed a tandem dimer. This construct, in which two EosFP subunits are connected by a flexible 12 amino acid linker, expresses well after fusion with the androgen and endothelin A receptors at 37°C. A variety of applications in cellular imaging, developmental biology and automated high-content screening applications are presented, which demonstrate that EosFP is a powerful tool for in vivo monitoring of cellular processes.
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