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Serial Femtosecond Crystallography Reveals that Photoactivation in a Fluorescent Protein Proceeds via the Hula Twist Mechanism

Year of publication

2023

Authors

Fadini, Alisia; Hutchison, Christopher D. M.; Morozov, Dmitry; Chang, Jeffrey; Maghlaoui, Karim; Perrett, Samuel; Luo, Fangjia; Kho, Jeslyn C. X.; Romei, Matthew G.; Morgan, R. Marc L.; Orr, Christian M.; Cordon-Preciado, Violeta; Fujiwara, Takaaki; Nuemket, Nipawan; Tosha, Takehiko; Tanaka, Rie; Owada, Shigeki; Tono, Kensuke; Iwata, So; Boxer, Steven G.; Groenhof, Gerrit; Nango, Eriko; van Thor, Jasper J.
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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.
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Organizations and authors

University of Jyväskylä

Morozov Dmitry Orcid -palvelun logo

Groenhof Gerrit Orcid -palvelun logo

Publication type

Publication format

Article

Parent publication type

Journal

Article type

Original article

Audience

Scientific

Peer-reviewed

Peer-Reviewed

MINEDU's publication type classification code

A1 Journal article (refereed), original research

Publication channel information

Volume

145

Issue

29

Pages

15796-15808

​Publication forum

61812

​Publication forum level

3

Open access

Open access in the publisher’s service

Yes

Open access of publication channel

Partially open publication channel

Self-archived

Yes

Other information

Fields of science

Chemical sciences

Keywords

[object Object],[object Object],[object Object]

Publication country

United States

Internationality of the publisher

International

Language

English

International co-publication

Yes

Co-publication with a company

No

DOI

10.1021/jacs.3c02313

The publication is included in the Ministry of Education and Culture’s Publication data collection

Yes