Photoconvertible fluorescent protein EosFP: biophysical properties and cell biology applications.
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.
Nienhaus
G Ulrich
GU
Department of Biophysics, University of Ulm, Ulm, Germany. uli@uiuc.edu
Nienhaus
Karin
K
Hölzle
Angela
A
Ivanchenko
Sergey
S
Renzi
Fabiana
F
Oswald
Franz
F
Wolff
Michael
M
Schmitt
Florian
F
Röcker
Carlheinz
C
Vallone
Beatrice
B
Weidemann
Wolfgang
W
Heilker
Ralf
R
Nar
Herbert
H
Wiedenmann
Jörg
J
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
D000596:Amino Acids; D000728:Androgens; D008164:Luminescent Proteins; D017466:Receptors, Endothelin