Targeted green-red photoconversion of EosFP, a fluorescent marker protein


Ivanchenko, S., Röcker, C., Oswald, F., Wiedenmann, J. and Nienhaus, G.U. (2005) Targeted green-red photoconversion of EosFP, a fluorescent marker protein. Journal of Biological Physics, 31, (3-4), 249-259. (doi:10.1007/s10867-005-0174-z).

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Description/Abstract

EosFP is a novel fluorescent protein from the stony coral Lobophyllia hemprichii. Its gene was cloned in Escherichia coli to express the tetrameric wild-type protein. The protein emits strong green fluorescence (516 nm) that shifts toward red (581 nm) upon near-ultraviolet irradiation at ∼390 nm due to a photo-induced modification that involves a break in the peptide backbone next to the chromophore. Using site-directed mutagenesis, dimeric (d1EosFP, d2EosFP) and monomeric (mEosFP) variants were produced with essentially unaltered spectroscopic properties. Here we present a spectroscopic characterization of EosFP and its variants, including room- and low-temperature spectra, fluorescence lifetime determinations, two-photon excitation and two-photon photoconversion. Furthermore, by transfection of a human cancer (HeLa) cell with a fusion construct of a mitochondrial targeting sequence and d2EosFP, we demonstrate how localized photoconversion of EosFP can be employed for resolving intracellular processes.

Item Type: Article
ISSNs: 0092-0606 (print)
Related URLs:
Keywords: fluorescent protein - fusion protein - two-photon excitation - fluorescence spectroscopy - photoconversion
Subjects: Q Science > QH Natural history > QH301 Biology
Divisions: University Structure - Pre August 2011 > School of Ocean & Earth Science (SOC/SOES)
ePrint ID: 51186
Date Deposited: 08 May 2008
Last Modified: 27 Mar 2014 18:34
URI: http://eprints.soton.ac.uk/id/eprint/51186

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