Molecular-Level Insight into the Spectral Tuning Mechanism of the DsRed Chromophore.
We present a detailed study of the protein environmental effects on the one- and two-photon absorption (1PA and 2PA, respectively) properties of the S0-S1 transition in the DsRed protein using the polarizable embedding density functional theory formalism. We find that steric factors and chromophore-protein interactions act in concert to enhance the 2PA activity inside the protein while adversely blue-shifting the 1PA maximum. A two-state model reveals that the 2PA intensity gain is primarily governed by the increased change in the permanent dipole moment between the ground and the excited states acquired inside the protein. Our results indicate that this mainly is attributable to counter-directional contributions stemming from Lys163 and the conserved Arg95 with the former additionally identified as a key residue in the color tuning mechanism. The results provide new insight into the tuning mechanism of DsRed and suggest a possible strategy for simultaneous improvement of its 1PA and 2PA properties.
†Department of Physics, Chemistry and Pharmacy, University of Southern Denmark, DK-5230 Odense M, Denmark.
Nanna H
NH
List
†Department of Physics, Chemistry and Pharmacy, University of Southern Denmark, DK-5230 Odense M, Denmark.
Jógvan Magnus H
JM
Olsen
†Department of Physics, Chemistry and Pharmacy, University of Southern Denmark, DK-5230 Odense M, Denmark.
Hans Jørgen Aa
HJ
Jensen
‡Centre of Theoretical and Computational Chemistry, Department of Chemistry, N-9037 Tromsø, Norway.
Arnfinn H
AH
Steindal
†Department of Physics, Chemistry and Pharmacy, University of Southern Denmark, DK-5230 Odense M, Denmark.
Jacob
J
Kongsted
TDDFT; fluorescent proteins; local electric field; polarizable embedding; polarization effects; spectral tuning; two-photon absorption