title: Photoconvertible fluorescent protein EosFP: biophysical properties and cell biology applications.
abstract: 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 degrees 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 degrees 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.
authors.affiliation: Department of Biophysics, University of Ulm, Ulm, Germany. uli@uiuc.edu
authors.forename: G Ulrich
authors.initials: GU
authors.lastname: Nienhaus
authors.forename: Karin
authors.initials: K
authors.lastname: Nienhaus
authors.forename: Angela
authors.initials: A
authors.lastname: Hölzle
authors.forename: Sergey
authors.initials: S
authors.lastname: Ivanchenko
authors.forename: Fabiana
authors.initials: F
authors.lastname: Renzi
authors.forename: Franz
authors.initials: F
authors.lastname: Oswald
authors.forename: Michael
authors.initials: M
authors.lastname: Wolff
authors.forename: Florian
authors.initials: F
authors.lastname: Schmitt
authors.forename: Carlheinz
authors.initials: C
authors.lastname: Röcker
authors.forename: Beatrice
authors.initials: B
authors.lastname: Vallone
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Weidemann
authors.forename: Ralf
authors.initials: R
authors.lastname: Heilker
authors.forename: Herbert
authors.initials: H
authors.lastname: Nar
authors.forename: Jörg
authors.initials: J
authors.lastname: Wiedenmann
mesh_terms: D000596:Amino Acids / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000728:Androgens / Q000235:genetics / Q000378:metabolism; D000818:Animals; D037421:Anthozoa / Q000737:chemistry*; D001709:Biotechnology / Q000379:methods*; D002468:Cell Physiological Phenomena; D018360:Crystallography, X-Ray; D019281:Dimerization; D008164:Luminescent Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D010777:Photochemistry; D017466:Receptors, Endothelin / Q000235:genetics / Q000378:metabolism; D014466:Ultraviolet Rays
chemical_list: D000596:Amino Acids; D000728:Androgens; D008164:Luminescent Proteins; D017466:Receptors, Endothelin