التفاصيل البيبلوغرافية
| العنوان: |
Mechanism and dynamics of fatty acid photodecarboxylase. |
| المؤلفون: |
Sorigué, D., Hadjidemetriou, K., Blangy, S., Gotthard, G., Bonvalet, A., Coquelle, N., Samire, P., Aleksandrov, A., Antonucci, L., Benachir, A., Boutet, S., Byrdin, M., Cammarata, M., Carbajo, S., Cuiné, S., Doak, R. B., Foucar, L., Gorel, A., Grünbein, M., Hartmann, E., Hienerwadel, R., Hilpert, M., Kloos, M., Lane, T. J., Légeret, B., Legrand, P., Li-Beisson, Y., Moulin, S. L. Y., Nurizzo, D., Peltier, G., Schirò, G., Shoeman, R. L., Sliwa, Michel, Solinas, X., Zhuang, B., Barends, T. R. M., Colletier, J. P., Joffre, M., Royant, A., Berthomieu, C., Weik, M., Domratcheva, T., Brettel, K., Vos, M. H., Schlichting, I., Arnoux, P., Müller, P., Beisson, F. |
| المساهمون: |
Université de Lille, CNRS, Institut de Biosciences et Biotechnologies d'Aix-Marseille (ex-IBEB) BIAM, Institut de biologie structurale IBS - UMR 5075, European Synchrotron Radiation Facility Grenoble ESRF, Laboratoire d'Optique et Biosciences LOB, Institut Polytechnique de Paris IP Paris, Institut de Biologie Intégrative de la Cellule I2BC, Institut de Physique de Rennes IPR, Laboratoire Avancé de Spectroscopie pour les Intéractions la Réactivité et l'Environnement (LASIRE) - UMR 8516 |
| سنة النشر: |
2024 |
| المجموعة: |
LillOA (Lille Open Archive - Université de Lille) |
| الوصف: |
INTRODUCTION Photoenzymes are rare biocatalysts driven by absorption of a photon at each catalytic cycle; they inspire development of artificial photoenzymes with valuable activities. Fatty acid photodecarboxylase (FAP) is a natural photoenzyme that has potential applications in the bio-based production of hydrocarbons, yet its mechanism is far from fully understood. RATIONALE To elucidate the mechanism of FAP, we studied the wild-type (WT) enzyme from Chlorella variabilis (CvFAP) and variants with altered active-site residues using a wealth of techniques, including static and time-resolved crystallography and spectroscopy, as well as biochemical and computational approaches. RESULTS A 1.8-Å-resolution CvFAP x-ray crystal structure revealed a dense hydrogen-bonding network positioning the fatty acid carboxyl group in the vicinity of the flavin adenine dinucleotide (FAD) cofactor. Structures solved from free electron laser and low-dose synchrotron x-ray crystal data further highlighted an unusual bent shape of the oxidized flavin chromophore, and showed that the bending angle (14°) did not change upon photon absorption (step 1) or throughout the photocycle. Calculations showed that bending substantially affected the energy levels of the flavin. Structural and spectroscopic analysis of WT and mutant proteins targeting two conserved active-site residues, R451 and C432, demonstrated that both residues were crucial for proper positioning of the substrate and water molecules and for oxidation of the fatty acid carboxylate by 1FAD* (~300 ps in WT FAP) to form FAD●– (step 2). Time-resolved infrared spectroscopy demonstrated that decarboxylation occured quasi-instantaneously upon this forward electron transfer, consistent with barrierless bond cleavage predicted by quantum chemistry calculations and with snapshots obtained by time-resolved crystallography. Transient absorption spectroscopy in H2O and D2O buffers indicated that back electron transfer from FAD●– was coupled to and limited by transfer of an exchangeable ... |
| نوع الوثيقة: |
article in journal/newspaper |
| وصف الملف: |
application/pdf |
| اللغة: |
English |
| Relation: |
Science; http://hdl.handle.net/20.500.12210/109533 |
| الاتاحة: |
https://hdl.handle.net/20.500.12210/109533 |
| Rights: |
info:eu-repo/semantics/openAccess |
| رقم الانضمام: |
edsbas.6DB3A57F |
| قاعدة البيانات: |
BASE |