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

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Resumo: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.
Autores principais:Vilela-Alves, Guilherme
Outros Autores:Manuel, Rita R.; Viegas, Aldino; Carpentier, Philippe; Biaso, Frédéric; Guigliarelli, Bruno; Pereira, Inês A.C.; Romão, Maria João; Mota, Cristiano
Assunto:General Chemistry SDG 13 - Climate Action
Ano:2024
País:Portugal
Tipo de documento:artigo
Tipo de acesso:acesso aberto
Instituição associada:Universidade Nova de Lisboa
Idioma:inglês
Origem:Repositório Institucional da UNL
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author Vilela-Alves, Guilherme
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
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
contributor_name_str_mv Faculdade de Ciências e Tecnologia (FCT)
UCIBIO - Applied Molecular Biosciences Unit
Instituto de Tecnologia Química e Biológica António Xavier (ITQB)
ROYAL SOC CHEMISTRY
RUN
country_str PT
creators_json_txt [{\"Person.name\":\"Vilela-Alves, Guilherme\"},{\"Person.name\":\"Manuel, Rita R.\"},{\"Person.name\":\"Viegas, Aldino\"},{\"Person.name\":\"Carpentier, Philippe\"},{\"Person.name\":\"Biaso, Frédéric\"},{\"Person.name\":\"Guigliarelli, Bruno\"},{\"Person.name\":\"Pereira, Inês A.C.\"},{\"Person.name\":\"Romão, Maria João\"},{\"Person.name\":\"Mota, Cristiano\"}]
datacite.contributors.contributor.contributorName.fl_str_mv Faculdade de Ciências e Tecnologia (FCT)
UCIBIO - Applied Molecular Biosciences Unit
Instituto de Tecnologia Química e Biológica António Xavier (ITQB)
ROYAL SOC CHEMISTRY
RUN
datacite.creators.creator.creatorName.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
datacite.date.Accepted.fl_str_mv 2024-08-14T00:00:00Z
datacite.date.available.fl_str_mv 2025-05-07T21:20:03Z
datacite.date.embargoed.fl_str_mv 2025-05-07T21:20:03Z
datacite.rights.fl_str_mv http://purl.org/coar/access_right/c_abf2
datacite.subjects.subject.fl_str_mv General Chemistry
SDG 13 - Climate Action
datacite.titles.title.fl_str_mv Substrate-dependent oxidative inactivation of a W-dependent formate dehydrogenase involving selenocysteine displacement
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)
ROYAL SOC CHEMISTRY
RUN
dc.creator.none.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.date.Accepted.fl_str_mv 2024-08-14T00:00:00Z
dc.date.available.fl_str_mv 2025-05-07T21:20:03Z
dc.date.embargoed.fl_str_mv 2025-05-07T21:20:03Z
dc.format.none.fl_str_mv application/pdf
dc.identifier.none.fl_str_mv http://hdl.handle.net/10362/182755
dc.language.none.fl_str_mv eng
dc.rights.none.fl_str_mv http://purl.org/coar/access_right/c_abf2
dc.subject.none.fl_str_mv General Chemistry
SDG 13 - Climate Action
dc.title.fl_str_mv Substrate-dependent oxidative inactivation of a W-dependent formate dehydrogenase involving selenocysteine displacement
dc.type.none.fl_str_mv http://purl.org/coar/resource_type/c_6501
description 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.
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eu_rights_str_mv openAccess
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fulltext.url.fl_str_mv https://run.unl.pt/bitstreams/162d15d1-e6fc-48bf-a11c-211656acae0c/download
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inst_facet_str urn:organizationAcronym:unl{{{_:::_}}}Universidade Nova de Lisboa
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instname_str Universidade Nova de Lisboa
language eng
network_acronym_str run
network_name_str Repositório Institucional da UNL
oai_identifier_str oai:run.unl.pt:10362/182755
organization_str_mv urn:organizationAcronym:unl
person_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
publishDate 2024
repo_facet_str urn:repositoryAcronym:run{{{_:::_}}}Repositório Institucional da UNL
reponame_str Repositório Institucional da UNL
repository_id_str urn:repositoryAcronym:run
service_str_mv urn:repositoryAcronym:run
spelling engenMetal-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.application/pdfenSubstrate-dependent oxidative inactivation of a W-dependent formate dehydrogenase involving selenocysteine displacementVilela-Alves, GuilhermeManuel, Rita R.Viegas, AldinoCarpentier, PhilippeBiaso, FrédéricGuigliarelli, BrunoPereira, Inês A.C.Romão, Maria JoãoMota, CristianoFaculdade de Ciências e Tecnologia (FCT)UCIBIO - Applied Molecular Biosciences UnitInstituto de Tecnologia Química e Biológica António Xavier (ITQB)ROYAL SOC CHEMISTRYHostingInstitutionOrganizationalRUNe-mailmailto:run@unl.ptrun@unl.ptISSNIsPartOf2041-6520URNIsPartOfPURE: 99997349URNIsPartOfPURE UUID: 81d2b972-3c1c-4a34-8cff-ce3dee6606a1URNIsPartOfScopus: 85199534626URNIsPartOfORCID: /0000-0002-3004-0543/work/183499873URNIsPartOfORCID: /0000-0002-8999-0420/work/183500013DOIIsPartOf10.1039/d4sc02394c2025-05-07T21:20:03Z2024-08-142024-08-14T00:00:00ZHandlehttp://hdl.handle.net/10362/182755http://purl.org/coar/access_right/c_abf2open accessGeneral ChemistrySDG 13 - Climate Action20001335 bytesliteraturehttp://purl.org/coar/resource_type/c_6501journal articlehttp://purl.org/coar/access_right/c_abf2application/pdffulltexthttps://run.unl.pt/bitstreams/162d15d1-e6fc-48bf-a11c-211656acae0c/download
spellingShingle Substrate-dependent oxidative inactivation of a W-dependent formate dehydrogenase involving selenocysteine displacement
Vilela-Alves, Guilherme
General Chemistry
SDG 13 - Climate Action
status SINGLETON
subject.fl_str_mv General Chemistry
SDG 13 - Climate Action
title 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_short 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
topic General Chemistry
SDG 13 - Climate Action
topic_facet General Chemistry
SDG 13 - Climate Action
url http://hdl.handle.net/10362/182755
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