Enhancement of Photophysical and Photosensitizing properties of Flavin Adenine Dinucleotide by Mutagenesis of the C-terminal extension of a bacterial Flavodoxin Reductase

Autores
Valle, Lorena; Abatedaga, Inés; Moran Vieyra, Faustino Eduardo; Bortolotti, Ana; Cortez, Nestor Ricardo; Borsarelli, Claudio Darío
Año de publicación
2015
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
The role of the mobile C-terminal extension present in Rhodobacter capsulatus ferredoxin–NADP(H) reductase (RcFPR) was evaluated using steady-state and dynamic spectroscopies for both intrinsic Trp and FAD in a series of mutants in the absence of NADP(H). Deletion of the six C-terminal amino acids beyond Ala266 was combined with the replacement A266Y to emulate the structure of plastidic reductases. Our results show that these modifications of the wild-type RcFPR produce subtle global conformational changes, but strongly reduce the local rigidity of the FAD-binding pocket, exposing the isoalloxazine ring to the solvent. Thus, the ultrafast charge-transfer quenching of 1 FAD* by the conserved Tyr66 residue was absent in the mutant series, producing enhancement of the excited singletand triplet-state properties of FAD. This work highlights the delicate balance of the specific interactions between FAD and the surrounding amino acids, and how the functionality and/or photostability of redox flavoproteins can be modified.
Fil: Valle, Lorena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Santiago del Estero; Argentina
Fil: Abatedaga, Inés. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Santiago del Estero; Argentina
Fil: Moran Vieyra, Faustino Eduardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Santiago del Estero; Argentina
Fil: Bortolotti, Ana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Biología Molecular y Celular de Rosario; Argentina
Fil: Cortez, Nestor Ricardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Biología Molecular y Celular de Rosario; Argentina
Fil: Borsarelli, Claudio Darío. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Santiago del Estero; Argentina
Materia
Flavoprotein
Fluorescence
Oxidoreductase
Singlet Oxygen
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:11336/13439
Acceder
id CONICETDig_32e961651526af5707206df5347c52c4
oai_identifier_str oai:ri.conicet.gov.ar:11336/13439
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Enhancement of Photophysical and Photosensitizing properties of Flavin Adenine Dinucleotide by Mutagenesis of the C-terminal extension of a bacterial Flavodoxin ReductaseValle, LorenaAbatedaga, InésMoran Vieyra, Faustino EduardoBortolotti, AnaCortez, Nestor RicardoBorsarelli, Claudio DaríoFlavoproteinFluorescenceOxidoreductaseSinglet Oxygenhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1https://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1The role of the mobile C-terminal extension present in Rhodobacter capsulatus ferredoxin–NADP(H) reductase (RcFPR) was evaluated using steady-state and dynamic spectroscopies for both intrinsic Trp and FAD in a series of mutants in the absence of NADP(H). Deletion of the six C-terminal amino acids beyond Ala266 was combined with the replacement A266Y to emulate the structure of plastidic reductases. Our results show that these modifications of the wild-type RcFPR produce subtle global conformational changes, but strongly reduce the local rigidity of the FAD-binding pocket, exposing the isoalloxazine ring to the solvent. Thus, the ultrafast charge-transfer quenching of 1 FAD* by the conserved Tyr66 residue was absent in the mutant series, producing enhancement of the excited singletand triplet-state properties of FAD. This work highlights the delicate balance of the specific interactions between FAD and the surrounding amino acids, and how the functionality and/or photostability of redox flavoproteins can be modified.Fil: Valle, Lorena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Santiago del Estero; ArgentinaFil: Abatedaga, Inés. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Santiago del Estero; ArgentinaFil: Moran Vieyra, Faustino Eduardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Santiago del Estero; ArgentinaFil: Bortolotti, Ana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Biología Molecular y Celular de Rosario; ArgentinaFil: Cortez, Nestor Ricardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Biología Molecular y Celular de Rosario; ArgentinaFil: Borsarelli, Claudio Darío. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Santiago del Estero; ArgentinaWiley Vch Verlag2015-03info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/13439Valle, Lorena; Abatedaga, Inés; Moran Vieyra, Faustino Eduardo; Bortolotti, Ana; Cortez, Nestor Ricardo; et al.; Enhancement of Photophysical and Photosensitizing properties of Flavin Adenine Dinucleotide by Mutagenesis of the C-terminal extension of a bacterial Flavodoxin Reductase; Wiley Vch Verlag; Chemphyschem; 16; 4; 3-2015; 872–8831439-7641enginfo:eu-repo/semantics/altIdentifier/doi/10.1002/cphc.201402774info:eu-repo/semantics/altIdentifier/url/http://onlinelibrary.wiley.com/doi/10.1002/cphc.201402774/abstractinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-17T10:56:28Zoai:ri.conicet.gov.ar:11336/13439instacron: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:34982025-09-17 10:56:28.236CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Enhancement of Photophysical and Photosensitizing properties of Flavin Adenine Dinucleotide by Mutagenesis of the C-terminal extension of a bacterial Flavodoxin Reductase
title Enhancement of Photophysical and Photosensitizing properties of Flavin Adenine Dinucleotide by Mutagenesis of the C-terminal extension of a bacterial Flavodoxin Reductase
spellingShingle Enhancement of Photophysical and Photosensitizing properties of Flavin Adenine Dinucleotide by Mutagenesis of the C-terminal extension of a bacterial Flavodoxin Reductase
Valle, Lorena
Flavoprotein
Fluorescence
Oxidoreductase
Singlet Oxygen
title_short Enhancement of Photophysical and Photosensitizing properties of Flavin Adenine Dinucleotide by Mutagenesis of the C-terminal extension of a bacterial Flavodoxin Reductase
title_full Enhancement of Photophysical and Photosensitizing properties of Flavin Adenine Dinucleotide by Mutagenesis of the C-terminal extension of a bacterial Flavodoxin Reductase
title_fullStr Enhancement of Photophysical and Photosensitizing properties of Flavin Adenine Dinucleotide by Mutagenesis of the C-terminal extension of a bacterial Flavodoxin Reductase
title_full_unstemmed Enhancement of Photophysical and Photosensitizing properties of Flavin Adenine Dinucleotide by Mutagenesis of the C-terminal extension of a bacterial Flavodoxin Reductase
title_sort Enhancement of Photophysical and Photosensitizing properties of Flavin Adenine Dinucleotide by Mutagenesis of the C-terminal extension of a bacterial Flavodoxin Reductase
dc.creator.none.fl_str_mv Valle, Lorena
Abatedaga, Inés
Moran Vieyra, Faustino Eduardo
Bortolotti, Ana
Cortez, Nestor Ricardo
Borsarelli, Claudio Darío
author Valle, Lorena
author_facet Valle, Lorena
Abatedaga, Inés
Moran Vieyra, Faustino Eduardo
Bortolotti, Ana
Cortez, Nestor Ricardo
Borsarelli, Claudio Darío
author_role author
author2 Abatedaga, Inés
Moran Vieyra, Faustino Eduardo
Bortolotti, Ana
Cortez, Nestor Ricardo
Borsarelli, Claudio Darío
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Flavoprotein
Fluorescence
Oxidoreductase
Singlet Oxygen
topic Flavoprotein
Fluorescence
Oxidoreductase
Singlet Oxygen
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv The role of the mobile C-terminal extension present in Rhodobacter capsulatus ferredoxin–NADP(H) reductase (RcFPR) was evaluated using steady-state and dynamic spectroscopies for both intrinsic Trp and FAD in a series of mutants in the absence of NADP(H). Deletion of the six C-terminal amino acids beyond Ala266 was combined with the replacement A266Y to emulate the structure of plastidic reductases. Our results show that these modifications of the wild-type RcFPR produce subtle global conformational changes, but strongly reduce the local rigidity of the FAD-binding pocket, exposing the isoalloxazine ring to the solvent. Thus, the ultrafast charge-transfer quenching of 1 FAD* by the conserved Tyr66 residue was absent in the mutant series, producing enhancement of the excited singletand triplet-state properties of FAD. This work highlights the delicate balance of the specific interactions between FAD and the surrounding amino acids, and how the functionality and/or photostability of redox flavoproteins can be modified.
Fil: Valle, Lorena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Santiago del Estero; Argentina
Fil: Abatedaga, Inés. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Santiago del Estero; Argentina
Fil: Moran Vieyra, Faustino Eduardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Santiago del Estero; Argentina
Fil: Bortolotti, Ana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Biología Molecular y Celular de Rosario; Argentina
Fil: Cortez, Nestor Ricardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Biología Molecular y Celular de Rosario; Argentina
Fil: Borsarelli, Claudio Darío. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Santiago del Estero; Argentina
description The role of the mobile C-terminal extension present in Rhodobacter capsulatus ferredoxin–NADP(H) reductase (RcFPR) was evaluated using steady-state and dynamic spectroscopies for both intrinsic Trp and FAD in a series of mutants in the absence of NADP(H). Deletion of the six C-terminal amino acids beyond Ala266 was combined with the replacement A266Y to emulate the structure of plastidic reductases. Our results show that these modifications of the wild-type RcFPR produce subtle global conformational changes, but strongly reduce the local rigidity of the FAD-binding pocket, exposing the isoalloxazine ring to the solvent. Thus, the ultrafast charge-transfer quenching of 1 FAD* by the conserved Tyr66 residue was absent in the mutant series, producing enhancement of the excited singletand triplet-state properties of FAD. This work highlights the delicate balance of the specific interactions between FAD and the surrounding amino acids, and how the functionality and/or photostability of redox flavoproteins can be modified.
publishDate 2015
dc.date.none.fl_str_mv 2015-03
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/11336/13439
Valle, Lorena; Abatedaga, Inés; Moran Vieyra, Faustino Eduardo; Bortolotti, Ana; Cortez, Nestor Ricardo; et al.; Enhancement of Photophysical and Photosensitizing properties of Flavin Adenine Dinucleotide by Mutagenesis of the C-terminal extension of a bacterial Flavodoxin Reductase; Wiley Vch Verlag; Chemphyschem; 16; 4; 3-2015; 872–883
1439-7641
url http://hdl.handle.net/11336/13439
identifier_str_mv Valle, Lorena; Abatedaga, Inés; Moran Vieyra, Faustino Eduardo; Bortolotti, Ana; Cortez, Nestor Ricardo; et al.; Enhancement of Photophysical and Photosensitizing properties of Flavin Adenine Dinucleotide by Mutagenesis of the C-terminal extension of a bacterial Flavodoxin Reductase; Wiley Vch Verlag; Chemphyschem; 16; 4; 3-2015; 872–883
1439-7641
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1002/cphc.201402774
info:eu-repo/semantics/altIdentifier/url/http://onlinelibrary.wiley.com/doi/10.1002/cphc.201402774/abstract
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
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Wiley Vch Verlag
publisher.none.fl_str_mv Wiley Vch Verlag
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
_version_ 1843606227912753152
score 13.001348

Publicaciones similares