Serial Femtosecond Crystallography Reveals that Photoactivation in a Fluorescent Protein Proceeds via the Hula Twist Mechanism
Publiceringsår
2023
Upphovspersoner
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.
Visa merAbstrakt
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.
Visa merOrganisationer och upphovspersoner
Publikationstyp
Publikationsform
Artikel
Moderpublikationens typ
Tidning
Artikelstyp
En originalartikel
Målgrupp
VetenskapligKollegialt utvärderad
Kollegialt utvärderadUKM:s publikationstyp
A1 Originalartikel i en vetenskaplig tidskriftPublikationskanalens uppgifter
Förläggare
Volym
145
Nummer
29
Sidor
15796-15808
ISSN
Publikationsforum
Publikationsforumsnivå
3
Öppen tillgång
Öppen tillgänglighet i förläggarens tjänst
Ja
Öppen tillgång till publikationskanalen
Delvis öppen publikationskanal
Parallellsparad
Ja
Övriga uppgifter
Vetenskapsområden
Kemi
Nyckelord
[object Object],[object Object],[object Object]
Publiceringsland
Förenta staterna (USA)
Förlagets internationalitet
Internationell
Språk
engelska
Internationell sampublikation
Ja
Sampublikation med ett företag
Nej
DOI
10.1021/jacs.3c02313
Publikationen ingår i undervisnings- och kulturministeriets datainsamling
Ja