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Acta Crystallographica Section D Structural Biology
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Time-resolved serial femtosecond crystallography on fatty-acid photodecarboxylase: lessons learned

Authors: Hadjidemetriou, Kyprianos; Coquelle, Nicolas; Barends, Thomas; de Zitter, Elke; Schlichting, Ilme; Colletier, Jacques Philippe; Weik, Martin;

Time-resolved serial femtosecond crystallography on fatty-acid photodecarboxylase: lessons learned

Abstract

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, CO2 or bicarbonate, was not possible because of the limited data quality, which was assigned to specificities of the crystalline system. Further TR-SFX experiments on a different crystal form are required to identify the photoproducts and their movements during the catalytic cycle.

Country
France
Keywords

serial femtosecond crystallography, Light, [SDV.BBM.BS] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], structure-factor extrapolation, MESH: Spectrum Analysis, Crystallography, X-Ray, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, fatty-acid photodecarboxylase, photoenzymes, Crystallography, Molecular Biology/Structural Biology [q-bio.BM], MESH: Crystallography, Lasers, Spectrum Analysis, Fatty Acids, 500, 540, Research Papers, MESH: Light, MESH: Fatty Acids, time-resolved crystallography, X-Ray, MESH: Lasers

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citations
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
9
Top 10%
Average
Top 10%
Green
hybrid
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