Protein dynamics promote hydride tunnelling in substrate oxidation by aryl-alcohol oxidase

The temperature dependence of hydride transfer from the substrate to the N5 of the FAD cofactor during the reductive half-reaction of Pleurotus eryngii aryl-alcohol oxidase (AAO) is assessed here. Kinetic isotope effects on both the pre-steady state reduction of the enzyme and its steady-state kinet...

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Detalhes bibliográficos
Autores: Carro, Juan, Martínez-Julve, Marta, Medina, Milagros, Martínez, Angel T., Ferreira, Patricia
Tipo de documento: artigo
Estado:Versión aceptada para publicación
Data de publicação:2020
País:España
Recursos:Universidad de Zaragoza
Repositório:Zaguán. Repositorio Digital de la Universidad de Zaragoza
OAI Identifier:oai:zaguan.unizar.es:75424
Acesso em linha:http://zaguan.unizar.es/record/75424
Access Level:Acceso aberto
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spelling Protein dynamics promote hydride tunnelling in substrate oxidation by aryl-alcohol oxidaseCarro, JuanMartínez-Julve, MartaMedina, MilagrosMartínez, Angel T.Ferreira, PatriciaThe temperature dependence of hydride transfer from the substrate to the N5 of the FAD cofactor during the reductive half-reaction of Pleurotus eryngii aryl-alcohol oxidase (AAO) is assessed here. Kinetic isotope effects on both the pre-steady state reduction of the enzyme and its steady-state kinetics, with differently deuterated substrates, suggest an environmentally-coupled quantum-mechanical tunnelling process. Moreover, those kinetic data, along with the crystallographic structure of the enzyme in complex with a substrate analogue, indicate that AAO shows a pre-organized active site that would only require the approaching of the hydride donor and acceptor for the tunnelled transfer to take place. Modification of the enzyme''s active-site architecture by replacement of Tyr92, a residue establishing hydrophobic interactions with the substrate analogue in the crystal structure, in the Y92F, Y92L and Y92W variants resulted in different temperature dependence patterns that indicated a role of this residue in modulating the transfer reaction.2020info:eu-repo/semantics/articleinfo:eu-repo/semantics/acceptedVersionapplication/pdfhttp://zaguan.unizar.es/record/75424reponame:Zaguán. Repositorio Digital de la Universidad de Zaragozainstname:Universidad de ZaragozaInglésinfo:eu-repo/grantAgreement/EC/FP7/613549info:eu-repo/grantAgreement/EC/H2020/720297This project has received funding from the European Union’s Horizon 2020 research and innovation program under grant agreement No H2020 720297-EnzOx2info:eu-repo/grantAgreement/ES/MINECO/BIO2016-75183-Pinfo:eu-repo/grantAgreement/ES/MINECO/NOESIS-BIO2014-56388-Rinfo:eu-repo/semantics/openAccessoai:zaguan.unizar.es:754242026-05-29T13:59:51Z
dc.title.none.fl_str_mv Protein dynamics promote hydride tunnelling in substrate oxidation by aryl-alcohol oxidase
title Protein dynamics promote hydride tunnelling in substrate oxidation by aryl-alcohol oxidase
spellingShingle Protein dynamics promote hydride tunnelling in substrate oxidation by aryl-alcohol oxidase
Carro, Juan
title_short Protein dynamics promote hydride tunnelling in substrate oxidation by aryl-alcohol oxidase
title_full Protein dynamics promote hydride tunnelling in substrate oxidation by aryl-alcohol oxidase
title_fullStr Protein dynamics promote hydride tunnelling in substrate oxidation by aryl-alcohol oxidase
title_full_unstemmed Protein dynamics promote hydride tunnelling in substrate oxidation by aryl-alcohol oxidase
title_sort Protein dynamics promote hydride tunnelling in substrate oxidation by aryl-alcohol oxidase
dc.creator.none.fl_str_mv Carro, Juan
Martínez-Julve, Marta
Medina, Milagros
Martínez, Angel T.
Ferreira, Patricia
author Carro, Juan
author_facet Carro, Juan
Martínez-Julve, Marta
Medina, Milagros
Martínez, Angel T.
Ferreira, Patricia
author_role author
author2 Martínez-Julve, Marta
Medina, Milagros
Martínez, Angel T.
Ferreira, Patricia
author2_role author
author
author
author
description The temperature dependence of hydride transfer from the substrate to the N5 of the FAD cofactor during the reductive half-reaction of Pleurotus eryngii aryl-alcohol oxidase (AAO) is assessed here. Kinetic isotope effects on both the pre-steady state reduction of the enzyme and its steady-state kinetics, with differently deuterated substrates, suggest an environmentally-coupled quantum-mechanical tunnelling process. Moreover, those kinetic data, along with the crystallographic structure of the enzyme in complex with a substrate analogue, indicate that AAO shows a pre-organized active site that would only require the approaching of the hydride donor and acceptor for the tunnelled transfer to take place. Modification of the enzyme''s active-site architecture by replacement of Tyr92, a residue establishing hydrophobic interactions with the substrate analogue in the crystal structure, in the Y92F, Y92L and Y92W variants resulted in different temperature dependence patterns that indicated a role of this residue in modulating the transfer reaction.
publishDate 2020
dc.date.none.fl_str_mv 2020
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/acceptedVersion
format article
status_str acceptedVersion
dc.identifier.none.fl_str_mv http://zaguan.unizar.es/record/75424
url http://zaguan.unizar.es/record/75424
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv info:eu-repo/grantAgreement/EC/FP7/613549
info:eu-repo/grantAgreement/EC/H2020/720297
This project has received funding from the European Union’s Horizon 2020 research and innovation program under grant agreement No H2020 720297-EnzOx2
info:eu-repo/grantAgreement/ES/MINECO/BIO2016-75183-P
info:eu-repo/grantAgreement/ES/MINECO/NOESIS-BIO2014-56388-R
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv
publisher.none.fl_str_mv
dc.source.none.fl_str_mv reponame:Zaguán. Repositorio Digital de la Universidad de Zaragoza
instname:Universidad de Zaragoza
instname_str Universidad de Zaragoza
reponame_str Zaguán. Repositorio Digital de la Universidad de Zaragoza
collection Zaguán. Repositorio Digital de la Universidad de Zaragoza
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15.300719

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