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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1 March 2006
Photoconvertible Fluorescent Protein EosFP: Biophysical Properties and Cell Biology Applications
G. Ulrich Nienhaus,
Karin Nienhaus,
Angela Hölzle,
Sergey Ivanchenko,
Fabiana Renzi,
Franz Oswald,
Michael Wolff,
Florian Schmitt,
Carlheinz Röcker,
Beatrice Vallone,
Wolfgang Weidemann,
Ralf Heilker,
Herbert Nar,
Jörg Wiedenmann
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Photochemistry and Photobiology
Vol. 82 • No. 2
March 2006
Vol. 82 • No. 2
March 2006