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Uriarte, Lucas, Vitalle, Rafaelle, Nizinski, Stanizlaw, Hadjidemetriou, Kyprianos, Zala, Ninon, Lukacs, Andras, Greetham, Gregory M., Sazanovich, Igor, Weik, Martin, Ruckebusch, Cyril, Meech, Steve 
ORCID: https://orcid.org/0000-0001-5561-2782 and Sliwa, Michel
(2022)
Structural information about the trans-to-cis isomerization mechanism of the photoswitchable fluorescent protein rsEGFP2 revealed by multiscale infrared transient absorption.
The Journal of Physical Chemistry Letters, 13 (5).
1194–1202.
ISSN 1948-7185
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Abstract
RsEGFP2 is a reversibly photoswitchable fluorescent protein used in super-resolved optical microscopies, which can be toggled between a fluorescent On state and a nonfluorescent Off state. Previous time-resolved ultraviolet–visible spectroscopic studies have shown that the Off-to-On photoactivation extends over the femto- to millisecond time scale and involves two picosecond lifetime excited states and four ground state intermediates, reflecting a trans-to-cis excited state isomerization, a millisecond deprotonation, and protein structural reorganizations. Femto- to millisecond time-resolved multiple-probe infrared spectroscopy (TRMPS-IR) can reveal structural aspects of intermediate species. Here we apply TRMPS-IR to rsEGFP2 and implement a Savitzky–Golay derivative analysis to correct for baseline drift. The results reveal that a subpicosecond twisted excited state precursor controls the trans-to-cis isomerization and the chromophore reaches its final position in the protein pocket within 100 ps. A new step with a time constant of 42 ns is reported and assigned to structural relaxation of the protein that occurs prior to the deprotonation of the chromophore on the millisecond time scale.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | materials science(all),physical and theoretical chemistry ,/dk/atira/pure/subjectarea/asjc/2500 |
| Faculty \ School: | Faculty of Science > School of Chemistry |
| UEA Research Groups: | Faculty of Science > Research Groups > Chemistry of Light and Energy Faculty of Science > Research Groups > Centre for Photonics and Quantum Science |
| Related URLs: | |
| Depositing User: | LivePure Connector |
| Date Deposited: | 22 Apr 2022 09:30 |
| Last Modified: | 09 Feb 2023 13:50 |
| URI: | https://ueaeprints.uea.ac.uk/id/eprint/84733 |
| DOI: | 10.1021/acs.jpclett.1c02920 |
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