A hydrogen bond network in the active site of Anabaena ferredoxin-NADP + reductase modulates its catalytic efficiency

Ferredoxin-nicotinamide-adenine dinucleotide phosphate (NADP+) reductase (FNR) catalyses the production of reduced nicotinamide-adenine dinucleotide phosphate (NADPH) in photosynthetic organisms, where its flavin adenine dinucleotide (FAD) cofactor takes two electrons from two reduced ferredoxin (Fd...

ver descrição completa

Detalhes bibliográficos
Autores: Sánchez-Azqueta, Ana, Herguedas, Beatriz, Hurtado-Guerrero, R., Hervás, Manuel, Navarro, José A., Martínez-Júlvez, Marta, Medina, Milagros
Tipo de documento: artigo
Data de publicação:2014
País:España
Recursos:Consejo Superior de Investigaciones Científicas (CSIC)
Repositório:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/98121
Acesso em linha:http://hdl.handle.net/10261/98121
Access Level:Acceso aberto
Palavra-chave:Kinetic isotope effect 27
Site-directed mutagenesis
Charge-transfer complex
Catalytically competent interaction
Hydride transfer
Isoalloxazine:nicotinamide interaction
Ferredoxin-NADP+ reductase
Flavoenzyme
id ES_b15bffc4fdfadce7e5aa009fe5ff8a55
oai_identifier_str oai:digital.csic.es:10261/98121
network_acronym_str ES
network_name_str España
repository_id_str
spelling A hydrogen bond network in the active site of Anabaena ferredoxin-NADP + reductase modulates its catalytic efficiencySánchez-Azqueta, AnaHerguedas, BeatrizHurtado-Guerrero, R.Hervás, ManuelNavarro, José A.Martínez-Júlvez, MartaMedina, MilagrosKinetic isotope effect 27Site-directed mutagenesisCharge-transfer complexCatalytically competent interactionHydride transferIsoalloxazine:nicotinamide interactionFerredoxin-NADP+ reductaseFlavoenzymeFerredoxin-nicotinamide-adenine dinucleotide phosphate (NADP+) reductase (FNR) catalyses the production of reduced nicotinamide-adenine dinucleotide phosphate (NADPH) in photosynthetic organisms, where its flavin adenine dinucleotide (FAD) cofactor takes two electrons from two reduced ferredoxin (Fd) molecules in two sequential steps, and transfers them to NADP+ in a single hydride transfer (HT) step. Despite the good knowledge of this catalytic machinery, additional roles can still be envisaged for already reported key residues, and new features are added to residues not previously identified as having a particular role in the mechanism. Here, we analyse for the first time the role of Ser59 in Anabaena FNR, a residue suggested by recent theoretical simulations as putatively involved in competent binding of the coenzyme in the active site by cooperating with Ser80. We show that Ser59 indirectly modulates the geometry of the active site, the interaction with substrates and the electronic properties of the isoalloxazine ring, and in consequence the electron transfer (ET) and HT processes. Additionally, we revise the role of Tyr79 and Ser80, previously investigated in homologous enzymes from plants. Our results probe that the active site of FNR is tuned by a H-bond network that involves the side-chains of these residues and that results to critical optimal substrate binding, exchange of electrons and, particularly, competent disposition of the C4n (hydride acceptor/donor) of the nicotinamide moiety of the coenzyme during the reversible HT event. © 2013 Elsevier B.V.Peer ReviewedElsevier20142014info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501http://hdl.handle.net/10261/98121reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglésinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/981212026-05-22T06:33:51Z
dc.title.none.fl_str_mv A hydrogen bond network in the active site of Anabaena ferredoxin-NADP + reductase modulates its catalytic efficiency
title A hydrogen bond network in the active site of Anabaena ferredoxin-NADP + reductase modulates its catalytic efficiency
spellingShingle A hydrogen bond network in the active site of Anabaena ferredoxin-NADP + reductase modulates its catalytic efficiency
Sánchez-Azqueta, Ana
Kinetic isotope effect 27
Site-directed mutagenesis
Charge-transfer complex
Catalytically competent interaction
Hydride transfer
Isoalloxazine:nicotinamide interaction
Ferredoxin-NADP+ reductase
Flavoenzyme
title_short A hydrogen bond network in the active site of Anabaena ferredoxin-NADP + reductase modulates its catalytic efficiency
title_full A hydrogen bond network in the active site of Anabaena ferredoxin-NADP + reductase modulates its catalytic efficiency
title_fullStr A hydrogen bond network in the active site of Anabaena ferredoxin-NADP + reductase modulates its catalytic efficiency
title_full_unstemmed A hydrogen bond network in the active site of Anabaena ferredoxin-NADP + reductase modulates its catalytic efficiency
title_sort A hydrogen bond network in the active site of Anabaena ferredoxin-NADP + reductase modulates its catalytic efficiency
dc.creator.none.fl_str_mv Sánchez-Azqueta, Ana
Herguedas, Beatriz
Hurtado-Guerrero, R.
Hervás, Manuel
Navarro, José A.
Martínez-Júlvez, Marta
Medina, Milagros
author Sánchez-Azqueta, Ana
author_facet Sánchez-Azqueta, Ana
Herguedas, Beatriz
Hurtado-Guerrero, R.
Hervás, Manuel
Navarro, José A.
Martínez-Júlvez, Marta
Medina, Milagros
author_role author
author2 Herguedas, Beatriz
Hurtado-Guerrero, R.
Hervás, Manuel
Navarro, José A.
Martínez-Júlvez, Marta
Medina, Milagros
author2_role author
author
author
author
author
author
dc.subject.none.fl_str_mv Kinetic isotope effect 27
Site-directed mutagenesis
Charge-transfer complex
Catalytically competent interaction
Hydride transfer
Isoalloxazine:nicotinamide interaction
Ferredoxin-NADP+ reductase
Flavoenzyme
topic Kinetic isotope effect 27
Site-directed mutagenesis
Charge-transfer complex
Catalytically competent interaction
Hydride transfer
Isoalloxazine:nicotinamide interaction
Ferredoxin-NADP+ reductase
Flavoenzyme
description Ferredoxin-nicotinamide-adenine dinucleotide phosphate (NADP+) reductase (FNR) catalyses the production of reduced nicotinamide-adenine dinucleotide phosphate (NADPH) in photosynthetic organisms, where its flavin adenine dinucleotide (FAD) cofactor takes two electrons from two reduced ferredoxin (Fd) molecules in two sequential steps, and transfers them to NADP+ in a single hydride transfer (HT) step. Despite the good knowledge of this catalytic machinery, additional roles can still be envisaged for already reported key residues, and new features are added to residues not previously identified as having a particular role in the mechanism. Here, we analyse for the first time the role of Ser59 in Anabaena FNR, a residue suggested by recent theoretical simulations as putatively involved in competent binding of the coenzyme in the active site by cooperating with Ser80. We show that Ser59 indirectly modulates the geometry of the active site, the interaction with substrates and the electronic properties of the isoalloxazine ring, and in consequence the electron transfer (ET) and HT processes. Additionally, we revise the role of Tyr79 and Ser80, previously investigated in homologous enzymes from plants. Our results probe that the active site of FNR is tuned by a H-bond network that involves the side-chains of these residues and that results to critical optimal substrate binding, exchange of electrons and, particularly, competent disposition of the C4n (hydride acceptor/donor) of the nicotinamide moiety of the coenzyme during the reversible HT event. © 2013 Elsevier B.V.
publishDate 2014
dc.date.none.fl_str_mv 2014
2014
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
format article
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/98121
url http://hdl.handle.net/10261/98121
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
repository.mail.fl_str_mv
_version_ 1869416924217081856
score 15,81155

Registros relacionados