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Serial femtosecond crystallography reveals that photoactivation in a fluorescent protein proceeds via the hula twist mechanism

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Title: Serial femtosecond crystallography reveals that photoactivation in a fluorescent protein proceeds via the hula twist mechanism
Authors: Fadini, A
Hutchison, CDM
Morozov, D
Chang, J
Maghlaoui, K
Perrett, S
Luo, F
Kho, JCX
Romei, MG
Morgan, RML
Orr, CM
Cordon-Preciado, V
Fujiwara, T
Nuemket, N
Tosha, T
Tanaka, R
Owada, S
Tono, K
Iwata, S
Boxer, SG
Groenhof, G
Nango, E
Van Thor, JJ
Item Type: Journal Article
Abstract: Chromophore cis/trans photoisomerization is a fundamental process in chemistry and in the activation of many photosensitive proteins. A major task is understanding the effect of the protein environment on the efficiency and direction of this reaction compared to what is observed in the gas and solution phases. In this study, we set out to visualize the hula twist (HT) mechanism in a fluorescent protein, which is hypothesized to be the preferred mechanism in a spatially constrained binding pocket. We use a chlorine substituent to break the twofold symmetry of the embedded phenolic group of the chromophore and unambiguously identify the HT primary photoproduct. Through serial femtosecond crystallography, we then track the photoreaction from femtoseconds to the microsecond regime. We observe signals for the photoisomerization of the chromophore as early as 300 fs, obtaining the first experimental structural evidence of the HT mechanism in a protein on its femtosecond-to-picosecond timescale. We are then able to follow how chromophore isomerization and twisting lead to secondary structure rearrangements of the protein β-barrel across the time window of our measurements.
Issue Date: 26-Jul-2023
Date of Acceptance: 1-Jul-2023
URI: http://hdl.handle.net/10044/1/105310
DOI: 10.1021/jacs.3c02313
ISSN: 0002-7863
Publisher: American Chemical Society
Start Page: 15796
End Page: 15808
Journal / Book Title: Journal of the American Chemical Society
Volume: 145
Issue: 29
Copyright Statement: © 2023 The Authors. Published by American Chemical Society. This work is published under a CC BY licence.
Publication Status: Published
Online Publication Date: 2023-07-07
Appears in Collections:Molecular Biosciences
Faculty of Natural Sciences