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  Ultrafast dynamics in the excited state of green fluorescent protein (wt) studied by frequency-resolved femtosecond pump-probe spectroscopy

Winkler, K., Lindner, J., Subramaniam, V., Jovin, T. M., & Voehringer, P. (2002). Ultrafast dynamics in the excited state of green fluorescent protein (wt) studied by frequency-resolved femtosecond pump-probe spectroscopy. Physical Chemistry - Chemical Physics, 4, 1072-1081. Retrieved from http://pubs.rsc.org/en/Content/ArticlePDF/2002/CP/B108843B/2002-03-04.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0012-F4E7-2 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0029-C5C8-B
Genre: Journal Article

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 Creators:
Winkler, K.1, Author              
Lindner, J.1, Author              
Subramaniam, V.2, Author              
Jovin, T. M.2, Author              
Voehringer, P.1, Author              
Affiliations:
1Research Group of Biomolecular and Chemical Dynamics, MPI for biophysical chemistry, Max Planck Society, ou_578553              
2Department of Molecular Biology, MPI for biophysical chemistry, Max Planck Society, ou_578628              

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Free keywords: Photoactive yellow protein; Proton-transfer; Ectothiorhodospira-halophila; Picosecond fluorescence; Single molecules; Chromophore; Excitation; Microscopy; Lifetimes; Mutants; Cells
 Abstract: The time and frequency resolved optical response of wild-type green fluorescent protein (wt-GFP) has been measured at room temperature following 30fs, 400nm photo-excitation. In the wavelength range covering the stationary fluorescence spectrum of the protein, the stimulated emission rises on a time scale of roughly 20ps due to excited-state proton-transfer (ESPT). The rise can be described phenomenologically by a sum of two exponentials. A long-time isosbestic behavior on the blue edge of the stationary emission implies a barrier for ESPT which is significantly larger than thermal excitations. In addition, an instantaneous component to the stimulated emission appears within the time resolution of our experiment. This observation is indicative of nonvertical cross-well transitions that prepare the proton-transferred configuration of the excited state directly from the equilibrium geometry of the ground-state neutral species during photo-excitation. Finally, transient absorptions around 500nm and 650nm can be observed, which are attributed to transitions from different protonated forms of the excited-state of GFP to higher lying electronic configurations, Sn . The entire optical response of GFP is quantitatively simulated using a dynamic model that includes: (i) an energy-dependent rate coefficient for ESPT, (ii) intra- and intermolecular transfer of excess vibrational energy (IVR and VET), and (iii) an additional non-radiative decay pathway for the initially prepared FranckÅ|Condon state leading to internal conversion via motion along a torsional coordinate. In particular, the nonexponential nature of the ESPT originates from overlapping time scales of reactive and non-reactive elementary processes following optical excitation.

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Language(s): eng - English
 Dates: 2005-08-182002
 Publication Status: Published in print
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 Rev. Method: Peer
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Title: Physical Chemistry - Chemical Physics
Source Genre: Journal
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Pages: - Volume / Issue: 4 Sequence Number: - Start / End Page: 1072 - 1081 Identifier: -