Mechanistic Insights into Ferredoxin–NADP(H) Reductase Catalysis Involving the Conserved Glutamate in the Active Site

Autores
Dumit, Veronica Ines; Essigke, Timm; Cortez, Nestor Ricardo; Ullmann, G. Matthias
Año de publicación
2010
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Plant-type ferredoxin?NADP(H) reductases (FNRs) are flavoenzymes harboring one molecule of noncovalently bound flavin adenine dinucleotide that catalyze reversible reactions between obligatory one-electron carriers and obligatory two-electron carriers. A glutamate next to the Cterminus is strictly conserved in FNR and has been proposed to function as proton donor/acceptor during catalysis. However, experimental studies of this proposed function led to contradicting conclusions about the role of thisglutamate in the catalytic mechanism. In the present work, we study the titration behavior of the glutamate in the active site of FNR using theoretical methods. Protonation probabilities for maize FNR were computed for the reaction intermediates of the catalytic cycle by Poisson?Boltzmann electrostatic calculations and Metropolis Monte Carlo titration. The titration behavior of the highly conserved glutamate was found to vary depending on the bound substrates NADP(H) and ferredoxin and also on the redox states of these substrates and the flavin adenine dinucleotide. Our resultssupport the involvement of the glutamate in the FNR catalytic mechanism not only as a proton donor but also as a key residue for stabilizing and destabilizing reaction intermediates. On the basis of our findings, we propose a model rationalizing the function of the glutamate in the reaction cycle, which allows reinterpretation of previous experimental results.
Fil: Dumit, Veronica Ines. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; Argentina
Fil: Essigke, Timm. University of Bayreuth; Alemania
Fil: Cortez, Nestor Ricardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; Argentina
Fil: Ullmann, G. Matthias. University of Bayreuth; Alemania
Materia
PHOTOSYNTHESIS
FLAVOPROTEIN
NADPH
FERREDOXIN
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/275337
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/275337
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Mechanistic Insights into Ferredoxin–NADP(H) Reductase Catalysis Involving the Conserved Glutamate in the Active SiteDumit, Veronica InesEssigke, TimmCortez, Nestor RicardoUllmann, G. MatthiasPHOTOSYNTHESISFLAVOPROTEINNADPHFERREDOXINhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Plant-type ferredoxin?NADP(H) reductases (FNRs) are flavoenzymes harboring one molecule of noncovalently bound flavin adenine dinucleotide that catalyze reversible reactions between obligatory one-electron carriers and obligatory two-electron carriers. A glutamate next to the Cterminus is strictly conserved in FNR and has been proposed to function as proton donor/acceptor during catalysis. However, experimental studies of this proposed function led to contradicting conclusions about the role of thisglutamate in the catalytic mechanism. In the present work, we study the titration behavior of the glutamate in the active site of FNR using theoretical methods. Protonation probabilities for maize FNR were computed for the reaction intermediates of the catalytic cycle by Poisson?Boltzmann electrostatic calculations and Metropolis Monte Carlo titration. The titration behavior of the highly conserved glutamate was found to vary depending on the bound substrates NADP(H) and ferredoxin and also on the redox states of these substrates and the flavin adenine dinucleotide. Our resultssupport the involvement of the glutamate in the FNR catalytic mechanism not only as a proton donor but also as a key residue for stabilizing and destabilizing reaction intermediates. On the basis of our findings, we propose a model rationalizing the function of the glutamate in the reaction cycle, which allows reinterpretation of previous experimental results.Fil: Dumit, Veronica Ines. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; ArgentinaFil: Essigke, Timm. University of Bayreuth; AlemaniaFil: Cortez, Nestor Ricardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; ArgentinaFil: Ullmann, G. Matthias. University of Bayreuth; AlemaniaAcademic Press Ltd - Elsevier Science Ltd2010-04info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/275337Dumit, Veronica Ines; Essigke, Timm; Cortez, Nestor Ricardo; Ullmann, G. Matthias; Mechanistic Insights into Ferredoxin–NADP(H) Reductase Catalysis Involving the Conserved Glutamate in the Active Site; Academic Press Ltd - Elsevier Science Ltd; Journal of Molecular Biology; 397; 3; 4-2010; 814-8250022-2836CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/abs/pii/S002228361000118Xinfo:eu-repo/semantics/altIdentifier/doi/10.1016/j.jmb.2010.01.063info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-12-03T09:39:28Zoai:ri.conicet.gov.ar:11336/275337instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-12-03 09:39:28.765CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Mechanistic Insights into Ferredoxin–NADP(H) Reductase Catalysis Involving the Conserved Glutamate in the Active Site
title Mechanistic Insights into Ferredoxin–NADP(H) Reductase Catalysis Involving the Conserved Glutamate in the Active Site
spellingShingle Mechanistic Insights into Ferredoxin–NADP(H) Reductase Catalysis Involving the Conserved Glutamate in the Active Site
Dumit, Veronica Ines
PHOTOSYNTHESIS
FLAVOPROTEIN
NADPH
FERREDOXIN
title_short Mechanistic Insights into Ferredoxin–NADP(H) Reductase Catalysis Involving the Conserved Glutamate in the Active Site
title_full Mechanistic Insights into Ferredoxin–NADP(H) Reductase Catalysis Involving the Conserved Glutamate in the Active Site
title_fullStr Mechanistic Insights into Ferredoxin–NADP(H) Reductase Catalysis Involving the Conserved Glutamate in the Active Site
title_full_unstemmed Mechanistic Insights into Ferredoxin–NADP(H) Reductase Catalysis Involving the Conserved Glutamate in the Active Site
title_sort Mechanistic Insights into Ferredoxin–NADP(H) Reductase Catalysis Involving the Conserved Glutamate in the Active Site
dc.creator.none.fl_str_mv Dumit, Veronica Ines
Essigke, Timm
Cortez, Nestor Ricardo
Ullmann, G. Matthias
author Dumit, Veronica Ines
author_facet Dumit, Veronica Ines
Essigke, Timm
Cortez, Nestor Ricardo
Ullmann, G. Matthias
author_role author
author2 Essigke, Timm
Cortez, Nestor Ricardo
Ullmann, G. Matthias
author2_role author
author
author
dc.subject.none.fl_str_mv PHOTOSYNTHESIS
FLAVOPROTEIN
NADPH
FERREDOXIN
topic PHOTOSYNTHESIS
FLAVOPROTEIN
NADPH
FERREDOXIN
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Plant-type ferredoxin?NADP(H) reductases (FNRs) are flavoenzymes harboring one molecule of noncovalently bound flavin adenine dinucleotide that catalyze reversible reactions between obligatory one-electron carriers and obligatory two-electron carriers. A glutamate next to the Cterminus is strictly conserved in FNR and has been proposed to function as proton donor/acceptor during catalysis. However, experimental studies of this proposed function led to contradicting conclusions about the role of thisglutamate in the catalytic mechanism. In the present work, we study the titration behavior of the glutamate in the active site of FNR using theoretical methods. Protonation probabilities for maize FNR were computed for the reaction intermediates of the catalytic cycle by Poisson?Boltzmann electrostatic calculations and Metropolis Monte Carlo titration. The titration behavior of the highly conserved glutamate was found to vary depending on the bound substrates NADP(H) and ferredoxin and also on the redox states of these substrates and the flavin adenine dinucleotide. Our resultssupport the involvement of the glutamate in the FNR catalytic mechanism not only as a proton donor but also as a key residue for stabilizing and destabilizing reaction intermediates. On the basis of our findings, we propose a model rationalizing the function of the glutamate in the reaction cycle, which allows reinterpretation of previous experimental results.
Fil: Dumit, Veronica Ines. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; Argentina
Fil: Essigke, Timm. University of Bayreuth; Alemania
Fil: Cortez, Nestor Ricardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; Argentina
Fil: Ullmann, G. Matthias. University of Bayreuth; Alemania
description Plant-type ferredoxin?NADP(H) reductases (FNRs) are flavoenzymes harboring one molecule of noncovalently bound flavin adenine dinucleotide that catalyze reversible reactions between obligatory one-electron carriers and obligatory two-electron carriers. A glutamate next to the Cterminus is strictly conserved in FNR and has been proposed to function as proton donor/acceptor during catalysis. However, experimental studies of this proposed function led to contradicting conclusions about the role of thisglutamate in the catalytic mechanism. In the present work, we study the titration behavior of the glutamate in the active site of FNR using theoretical methods. Protonation probabilities for maize FNR were computed for the reaction intermediates of the catalytic cycle by Poisson?Boltzmann electrostatic calculations and Metropolis Monte Carlo titration. The titration behavior of the highly conserved glutamate was found to vary depending on the bound substrates NADP(H) and ferredoxin and also on the redox states of these substrates and the flavin adenine dinucleotide. Our resultssupport the involvement of the glutamate in the FNR catalytic mechanism not only as a proton donor but also as a key residue for stabilizing and destabilizing reaction intermediates. On the basis of our findings, we propose a model rationalizing the function of the glutamate in the reaction cycle, which allows reinterpretation of previous experimental results.
publishDate 2010
dc.date.none.fl_str_mv 2010-04
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/275337
Dumit, Veronica Ines; Essigke, Timm; Cortez, Nestor Ricardo; Ullmann, G. Matthias; Mechanistic Insights into Ferredoxin–NADP(H) Reductase Catalysis Involving the Conserved Glutamate in the Active Site; Academic Press Ltd - Elsevier Science Ltd; Journal of Molecular Biology; 397; 3; 4-2010; 814-825
0022-2836
CONICET Digital
CONICET
url http://hdl.handle.net/11336/275337
identifier_str_mv Dumit, Veronica Ines; Essigke, Timm; Cortez, Nestor Ricardo; Ullmann, G. Matthias; Mechanistic Insights into Ferredoxin–NADP(H) Reductase Catalysis Involving the Conserved Glutamate in the Active Site; Academic Press Ltd - Elsevier Science Ltd; Journal of Molecular Biology; 397; 3; 4-2010; 814-825
0022-2836
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/abs/pii/S002228361000118X
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.jmb.2010.01.063
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Academic Press Ltd - Elsevier Science Ltd
publisher.none.fl_str_mv Academic Press Ltd - Elsevier Science Ltd
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.214268

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