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Ashworth, Eleanor K., Dezalay, Jordan, Ryan, Christopher, Ieritano, Christian, Hopkins, W. Scott, Chambrier, Isabelle, Cammidge, Andrew N. 
ORCID: https://orcid.org/0000-0001-7912-4310, Stockett, Mark H., Noble, Jennifer A. and Bull, James N. ORCID: https://orcid.org/0000-0003-0953-1716
(2023)
Protomers of the green and cyan fluorescent protein chromophores investigated using action spectroscopy.
Physical Chemistry Chemical Physics, 25 (30).
pp. 20405-20413.
ISSN 1463-9076
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Abstract
The photophysics of biochromophore ions often depends on the isomeric or protomeric distribution, yet this distribution, and the individual isomer contributions to an action spectrum, can be difficult to quantify. Here, we use two separate photodissociation action spectroscopy instruments to record electronic spectra for protonated forms of the green (pHBDI+) and cyan (Cyan+) fluorescent protein chromophores. One instrument allows for cryogenic (T = 40±10 K) cooling of the ions, while the other offers the ability to perform protomer-selective photodissociation spectroscopy. We show that both chromophores are generated as two protomers when using electrospray ionisation, and that the protomers have partially overlapping absorption profiles associated with the S1 ← S0 transition. The action spectra for both species span the 340–460 nm range, although the spectral onset for the pHBDI+ protomer with the proton residing on the carbonyl oxygen is red-shifted by ≈40 nm relative to the lower-energy imine protomer. Similarly, the imine and carbonyl protomers are the lowest energy forms of Cyan+, with the main band for the carbonyl protomer red-shifted by ≈60 nm relative to the lower-energy imine protomer. The present strategy for investigating protomers can be applied to a wide range of other biochromophore ions.
| Item Type: | Article |
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| Additional Information: | Acknowledgements: JNB acknowledges funding from an EPSRC New Investigator Award (EP/W018691). JAN acknowledges funding from the Agence Nationale de la Recherche (ANR, HYDRAE project ANR21-CE30-0004-01) and additional support from the French Programme National “Physique et Chimie du Milieu Interstellaire" (PCMI) of the CNRS/INSU with the INC/INP, co-funded by the CEA and the CNES. WSH acknowledges financial support provided by the Natural Sciences and Engineering Research Council (NSERC) of Canada in the form of Discovery and Alliance grants, and Mitacs in the form of an Accelerate grant. EKA thanks the University of East Anglia for a doctoral studentship. Travel funding was provided by the Swedish Foundation for International Cooperation in Research and Higher Education (STINT) Grant for Internationalisation programme (PT2017-7328 to MHS and JNB). Electronic structure calculations were carried out on the High Performance Computing Cluster supported by the Research and Specialist Computing Support service at the University of East Anglia |
| Uncontrolled Keywords: | physics and astronomy(all),physical and theoretical chemistry ,/dk/atira/pure/subjectarea/asjc/3100 |
| Faculty \ School: | Faculty of Science > School of Chemistry |
| 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 |
| Related URLs: | |
| Depositing User: | LivePure Connector |
| Date Deposited: | 14 Jul 2023 08:30 |
| Last Modified: | 29 Aug 2023 09:31 |
| URI: | https://ueaeprints.uea.ac.uk/id/eprint/92612 |
| DOI: | 10.1039/D3CP02661B |
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