Time-resolved serial femtosecond crystallography on fatty-acid photodecarboxylase: lessons learned.
fatty-acid photodecarboxylase
photoenzymes
serial femtosecond crystallography
structure-factor extrapolation
time-resolved crystallography
Journal
Acta crystallographica. Section D, Structural biology
ISSN: 2059-7983
Titre abrégé: Acta Crystallogr D Struct Biol
Pays: United States
ID NLM: 101676043
Informations de publication
Date de publication:
01 Sep 2022
01 Sep 2022
Historique:
received:
26
02
2022
accepted:
22
07
2022
entrez:
1
9
2022
pubmed:
2
9
2022
medline:
8
9
2022
Statut:
ppublish
Résumé
Upon absorption of a blue-light photon, fatty-acid photodecarboxylase catalyzes the decarboxylation of free fatty acids to form hydrocarbons (for example alkanes or alkenes). The major components of the catalytic mechanism have recently been elucidated by combining static and time-resolved serial femtosecond crystallography (TR-SFX), time-resolved vibrational and electronic spectroscopies, quantum-chemical calculations and site-directed mutagenesis [Sorigué et al. (2021), Science, 372, eabd5687]. The TR-SFX experiments, which were carried out at four different picosecond to microsecond pump-probe delays, yielded input for the calculation of Fourier difference maps that demonstrated light-induced decarboxylation. Here, some of the difficulties encountered during the experiment as well as during data processing are highlighted, in particular regarding space-group assignment, a pump-laser power titration is described and data analysis is extended by structure-factor extrapolation of the TR-SFX data. Structure refinement against extrapolated structure factors reveals a reorientation of the generated hydrocarbon and the formation of a photoproduct close to Cys432 and Arg451. Identification of its chemical nature, CO
Identifiants
pubmed: 36048153
pii: S2059798322007525
doi: 10.1107/S2059798322007525
pmc: PMC9435596
doi:
Substances chimiques
Fatty Acids
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
1131-1142Subventions
Organisme : European Research Council, H2020 European Research Council
ID : 724362
Informations de copyright
open access.
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