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ToolsFatima, Anam, Ashworth, Eleanor K., Fernandes, Isabelle, Cammidge, Andrew N., Bressan, Giovanni, Meech, Steve R. and Bull, James N. (2025) Ultrafast photophysics of the cyan fluorescent protein chromophore in solution. Physical Chemistry Chemical Physics, 27 (18). pp. 9407-9416. ISSN 1463-9076
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Abstract
Incorporation of fluorescent proteins (FPs) into biological systems has revolutionised bioimaging and the understanding of cellular processes. Ongoing developments of FPs are driving efforts to characterise the fundamental photoactive unit (chromophore) embedded within the protein. Cyan FP has a blue emitting chromophore and is widely used in Förster resonance energy transfer studies. Here, we probe the ultrafast photophysics of the cyan FP chromophore in solution using time-resolved fluorescence up-conversion and transient absorption spectroscopies. The ultrafast dynamics are characterised by two lifetimes, sub-picosecond τ 1 (or τ F) associated with loss of the fluorescent Franck-Condon state, and lifetime τ 2 on the order of several picoseconds that is linked with cooling of a hot ground state. MRSF-TDDFT calculations show that the relaxed S 1 state equilibrium geometry is classified as a partial twisted intramolecular charge-transfer state, and lies close in energy to a conical intersection seam associated with torsion about the central double bond leading to facile internal conversion. The excited state dynamics exhibit only a weak viscosity dependence, consistent with a barrierless and near-volume-conserving non-radiative decay mechanism. Fluorescence lifetimes for the deprotonated anion are twice those for the neutral.
| Item Type: | Article |
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| Additional Information: | Funding information: This work was funded by an EPSRC New Investigator Award (EP/W018691 to JNB) and an EPSRC Grant (EP/X011410/1 to SRM). EKA thanks the University of East Anglia for a doctoral scholarship. GB is grateful to the Leverhulme Trust for funding him through an Early Career Fellowship (grant no. ECF-2023-195). |
| Faculty \ School: | Faculty of Science > School of Chemistry, Pharmacy and Pharmacology Faculty of Science |
| UEA Research Groups: | Faculty of Science > Research Groups > Centre for Photonics and Quantum Science Faculty of Science > Research Groups > Chemistry of Materials and Catalysis Faculty of Science > Research Groups > Chemistry of Light and Energy |
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| Depositing User: | LivePure Connector |
| Date Deposited: | 15 Apr 2025 09:30 |
| Last Modified: | 14 May 2025 03:30 |
| URI: | https://ueaeprints.uea.ac.uk/id/eprint/99036 |
| DOI: | 10.1039/D5CP00942A |
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