Substrate-dependent oxidative inactivation of a W-dependent formate dehydrogenase involving selenocysteine displacement

Bibliographic Details
Main Author: Vilela-Alves, Guilherme
Publication Date: 2024
Other Authors: Manuel, Rita R., Viegas, Aldino, Carpentier, Philippe, Biaso, Frédéric, Guigliarelli, Bruno, Pereira, Inês A.C., Romão, Maria João, Mota, Cristiano
Format: Article
Language: eng
Source: Repositórios Científicos de Acesso Aberto de Portugal (RCAAP)
Download full: http://hdl.handle.net/10362/182755
Summary: Funding Information: This work was financially supported by Funda\u00E7\u00E3o para a Ci\u00EAncia e Tecnologia (FCT, Portugal) through fellowship no. 2023.00286.BD (G. V.-A.) and DFA/BD/7897/2020 (R. R. M.), grant no. PTDC/BII-BBF/2050/2020 (http://doi.org/10.54499/PTDC/BII-BBF/2050/2020) (I. A. C. P. and M. J. R.), research contract 2020.00043.CEECIND (A. V.) and R&D units MOSTMICRO-ITQB (grant no. UIDB/04612/2020 and UIDP/04612/2020) (I. A. C. P.) and UCIBIO (grant no. UIDP/04378/2020 and UIDB/04378/2020) (M. J. R.), and Associated Laboratories LS4FUTURE (grant no. LA/P/0087/2020) (I. A. C. P.) and i4HB (grant no. LA/P/0140/2020) (M. J. R.). The NMR spectrometers at FCT-NOVA are part of Rede Nacional de RMN (PTNMR), supported by FCT-MCTES (ROTEIRO/0031/2013-PINFRA/22161/2016) (cofinanced by FEDER through COMPETE 2020, POCI and PORL and FCT through PIDDAC). This work was also funded by the French national research agency (ANR \u2013 MOLYERE project, grant no. 16-CE-29- 0010-01) (B. G.). We thank Prof Carlos Rom\u00E3o for insightful discussions pertaining the coordination chemistry of the SeCys unbound active site and the excellent technical assistance of Jo\u00E3o Carita from ITQB NOVA on microbial cell growth. We thank the reviewers for their constructive comments that significantly enhanced the manuscript. We are also grateful to the EPR-MRS facilities of the Aix-Marseille University EPR centre and acknowledge the support of the European research infrastructure MOSBRI (grant no. 101004806) (B. G.) and the French research infrastructure INFRANALYTICS (MOLYERE, 16-CE-29-0010-01) (B. G.). This work was supported by the computing facilities of the CRCMM, \u201CCentre R\u00E9gional de Comp\u00E9tences en Mod\u00E9lisation Mol\u00E9culaire de Marseille\u201D. We also acknowledge the ESRF Synchrotron for provision of synchrotron radiation facilities, and we thank the staff of the ESRF and EMBL Grenoble for assistance and support in using the HPMX lab and beamlines ID23-1, ID30A-3, ID30B. Publisher Copyright: © 2024 The Royal Society of Chemistry.
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spelling Substrate-dependent oxidative inactivation of a W-dependent formate dehydrogenase involving selenocysteine displacementChemistry(all)SDG 13 - Climate ActionFunding Information: This work was financially supported by Funda\u00E7\u00E3o para a Ci\u00EAncia e Tecnologia (FCT, Portugal) through fellowship no. 2023.00286.BD (G. V.-A.) and DFA/BD/7897/2020 (R. R. M.), grant no. PTDC/BII-BBF/2050/2020 (http://doi.org/10.54499/PTDC/BII-BBF/2050/2020) (I. A. C. P. and M. J. R.), research contract 2020.00043.CEECIND (A. V.) and R&D units MOSTMICRO-ITQB (grant no. UIDB/04612/2020 and UIDP/04612/2020) (I. A. C. P.) and UCIBIO (grant no. UIDP/04378/2020 and UIDB/04378/2020) (M. J. R.), and Associated Laboratories LS4FUTURE (grant no. LA/P/0087/2020) (I. A. C. P.) and i4HB (grant no. LA/P/0140/2020) (M. J. R.). The NMR spectrometers at FCT-NOVA are part of Rede Nacional de RMN (PTNMR), supported by FCT-MCTES (ROTEIRO/0031/2013-PINFRA/22161/2016) (cofinanced by FEDER through COMPETE 2020, POCI and PORL and FCT through PIDDAC). This work was also funded by the French national research agency (ANR \u2013 MOLYERE project, grant no. 16-CE-29- 0010-01) (B. G.). We thank Prof Carlos Rom\u00E3o for insightful discussions pertaining the coordination chemistry of the SeCys unbound active site and the excellent technical assistance of Jo\u00E3o Carita from ITQB NOVA on microbial cell growth. We thank the reviewers for their constructive comments that significantly enhanced the manuscript. We are also grateful to the EPR-MRS facilities of the Aix-Marseille University EPR centre and acknowledge the support of the European research infrastructure MOSBRI (grant no. 101004806) (B. G.) and the French research infrastructure INFRANALYTICS (MOLYERE, 16-CE-29-0010-01) (B. G.). This work was supported by the computing facilities of the CRCMM, \u201CCentre R\u00E9gional de Comp\u00E9tences en Mod\u00E9lisation Mol\u00E9culaire de Marseille\u201D. We also acknowledge the ESRF Synchrotron for provision of synchrotron radiation facilities, and we thank the staff of the ESRF and EMBL Grenoble for assistance and support in using the HPMX lab and beamlines ID23-1, ID30A-3, ID30B. Publisher Copyright: © 2024 The Royal Society of Chemistry.Metal-dependent formate dehydrogenases are very promising targets for enzyme optimization and design of bio-inspired catalysts for CO2 reduction, towards innovative strategies for climate change mitigation. For effective application of these enzymes, the catalytic mechanism must be better understood, and the molecular determinants clarified. Despite numerous studies, several doubts persist, namely regarding the role played by the possible dissociation of the SeCys ligand from the Mo/W active site. Additionally, the oxygen sensitivity of these enzymes must also be understood as it poses an important obstacle for biotechnological applications. This work presents a combined biochemical, spectroscopic, and structural characterization of Desulfovibrio vulgaris FdhAB (DvFdhAB) when exposed to oxygen in the presence of a substrate (formate or CO2). This study reveals that O2 inactivation is promoted by the presence of either substrate and involves forming a different species in the active site, captured in the crystal structures, where the SeCys ligand is displaced from tungsten coordination and replaced by a dioxygen or peroxide molecule. This form was reproducibly obtained and supports the conclusion that, although W-DvFdhAB can catalyse the oxidation of formate in the presence of oxygen for some minutes, it gets irreversibly inactivated after prolonged O2 exposure in the presence of either substrate.Faculdade de Ciências e Tecnologia (FCT)UCIBIO - Applied Molecular Biosciences UnitInstituto de Tecnologia Química e Biológica António Xavier (ITQB)RUNVilela-Alves, GuilhermeManuel, Rita R.Viegas, AldinoCarpentier, PhilippeBiaso, FrédéricGuigliarelli, BrunoPereira, Inês A.C.Romão, Maria JoãoMota, Cristiano2025-05-07T21:20:03Z2024-08-142024-08-14T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article12application/pdfhttp://hdl.handle.net/10362/182755eng2041-6520PURE: 99997349https://doi.org/10.1039/d4sc02394cinfo:eu-repo/semantics/openAccessreponame:Repositórios Científicos de Acesso Aberto de Portugal (RCAAP)instname:FCCN, serviços digitais da FCT – Fundação para a Ciência e a Tecnologiainstacron:RCAAP2025-05-19T01:39:22Zoai:run.unl.pt:10362/182755Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireinfo@rcaap.ptopendoar:https://opendoar.ac.uk/repository/71602025-05-29T07:13:47.226630Repositórios Científicos de Acesso Aberto de Portugal (RCAAP) - FCCN, serviços digitais da FCT – Fundação para a Ciência e a Tecnologiafalse
dc.title.none.fl_str_mv Substrate-dependent oxidative inactivation of a W-dependent formate dehydrogenase involving selenocysteine displacement
title Substrate-dependent oxidative inactivation of a W-dependent formate dehydrogenase involving selenocysteine displacement
spellingShingle Substrate-dependent oxidative inactivation of a W-dependent formate dehydrogenase involving selenocysteine displacement
Vilela-Alves, Guilherme
Chemistry(all)
SDG 13 - Climate Action
title_short Substrate-dependent oxidative inactivation of a W-dependent formate dehydrogenase involving selenocysteine displacement
title_full Substrate-dependent oxidative inactivation of a W-dependent formate dehydrogenase involving selenocysteine displacement
title_fullStr Substrate-dependent oxidative inactivation of a W-dependent formate dehydrogenase involving selenocysteine displacement
title_full_unstemmed Substrate-dependent oxidative inactivation of a W-dependent formate dehydrogenase involving selenocysteine displacement
title_sort Substrate-dependent oxidative inactivation of a W-dependent formate dehydrogenase involving selenocysteine displacement
author Vilela-Alves, Guilherme
author_facet Vilela-Alves, Guilherme
Manuel, Rita R.
Viegas, Aldino
Carpentier, Philippe
Biaso, Frédéric
Guigliarelli, Bruno
Pereira, Inês A.C.
Romão, Maria João
Mota, Cristiano
author_role author
author2 Manuel, Rita R.
Viegas, Aldino
Carpentier, Philippe
Biaso, Frédéric
Guigliarelli, Bruno
Pereira, Inês A.C.
Romão, Maria João
Mota, Cristiano
author2_role author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Faculdade de Ciências e Tecnologia (FCT)
UCIBIO - Applied Molecular Biosciences Unit
Instituto de Tecnologia Química e Biológica António Xavier (ITQB)
RUN
dc.contributor.author.fl_str_mv Vilela-Alves, Guilherme
Manuel, Rita R.
Viegas, Aldino
Carpentier, Philippe
Biaso, Frédéric
Guigliarelli, Bruno
Pereira, Inês A.C.
Romão, Maria João
Mota, Cristiano
dc.subject.por.fl_str_mv Chemistry(all)
SDG 13 - Climate Action
topic Chemistry(all)
SDG 13 - Climate Action
description Funding Information: This work was financially supported by Funda\u00E7\u00E3o para a Ci\u00EAncia e Tecnologia (FCT, Portugal) through fellowship no. 2023.00286.BD (G. V.-A.) and DFA/BD/7897/2020 (R. R. M.), grant no. PTDC/BII-BBF/2050/2020 (http://doi.org/10.54499/PTDC/BII-BBF/2050/2020) (I. A. C. P. and M. J. R.), research contract 2020.00043.CEECIND (A. V.) and R&D units MOSTMICRO-ITQB (grant no. UIDB/04612/2020 and UIDP/04612/2020) (I. A. C. P.) and UCIBIO (grant no. UIDP/04378/2020 and UIDB/04378/2020) (M. J. R.), and Associated Laboratories LS4FUTURE (grant no. LA/P/0087/2020) (I. A. C. P.) and i4HB (grant no. LA/P/0140/2020) (M. J. R.). The NMR spectrometers at FCT-NOVA are part of Rede Nacional de RMN (PTNMR), supported by FCT-MCTES (ROTEIRO/0031/2013-PINFRA/22161/2016) (cofinanced by FEDER through COMPETE 2020, POCI and PORL and FCT through PIDDAC). This work was also funded by the French national research agency (ANR \u2013 MOLYERE project, grant no. 16-CE-29- 0010-01) (B. G.). We thank Prof Carlos Rom\u00E3o for insightful discussions pertaining the coordination chemistry of the SeCys unbound active site and the excellent technical assistance of Jo\u00E3o Carita from ITQB NOVA on microbial cell growth. We thank the reviewers for their constructive comments that significantly enhanced the manuscript. We are also grateful to the EPR-MRS facilities of the Aix-Marseille University EPR centre and acknowledge the support of the European research infrastructure MOSBRI (grant no. 101004806) (B. G.) and the French research infrastructure INFRANALYTICS (MOLYERE, 16-CE-29-0010-01) (B. G.). This work was supported by the computing facilities of the CRCMM, \u201CCentre R\u00E9gional de Comp\u00E9tences en Mod\u00E9lisation Mol\u00E9culaire de Marseille\u201D. We also acknowledge the ESRF Synchrotron for provision of synchrotron radiation facilities, and we thank the staff of the ESRF and EMBL Grenoble for assistance and support in using the HPMX lab and beamlines ID23-1, ID30A-3, ID30B. Publisher Copyright: © 2024 The Royal Society of Chemistry.
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2025-05-07T21:20:03Z
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