Effects of fe2+/fe3+ binding to human frataxin and its d122y variant, as revealed by site-directed spin labeling (Sdsl) epr complemented by fluorescence and circular dichroism spec...

Autores
Doni, Davide; Passerini, Leonardo; Audran, Gérard; Marque, Sylvain R. A.; Schulz, Marvin; Santos, Javier; Costantini, Paola; Bortolus, Marco; Carbonera, Donatella
Año de publicación
2020
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Frataxin is a highly conserved protein whose deficiency results in the neurodegenerative disease Friederich’s ataxia. Frataxin’s actual physiological function has been debated for a long time without reaching a general agreement; however, it is commonly accepted that the protein is involved in the biosynthetic iron-sulphur cluster (ISC) machinery, and several authors have pointed out that it also participates in iron homeostasis. In this work, we use site-directed spin labeling coupled to electron paramagnetic resonance (SDSL EPR) to add new information on the effects of ferric and ferrous iron binding on the properties of human frataxin in vitro. Using SDSL EPR and relating the results to fluorescence experiments commonly performed to study iron binding to FXN, we produced evidence that ferric iron causes reversible aggregation without preferred interfaces in a concentration-dependent fashion, starting at relatively low concentrations (micromolar range), whereas ferrous iron binds without inducing aggregation. Moreover, our experiments show that the ferrous binding does not lead to changes of protein conformation. The data reported in this study reveal that the currently reported binding stoichiometries should be taken with caution. The use of a spin label resistant to reduction, as well as the comparison of the binding effect of Fe2+ in wild type and in the pathological D122Y variant of frataxin, allowed us to characterize the Fe2+ binding properties of different protein sites and highlight the effect of the D122Y substitution on the surrounding residues. We suggest that both Fe2+ and Fe3+ might play a relevant role in the context of the proposed FXN physiological functions.
Fil: Doni, Davide. Università di Padova; Italia
Fil: Passerini, Leonardo. Università di Padova; Italia
Fil: Audran, Gérard. Centre national de la recherche scientifique. Institut de Chimie Radicalaire; Francia
Fil: Marque, Sylvain R. A.. Centre national de la recherche scientifique. Institut de Chimie Radicalaire; Francia
Fil: Schulz, Marvin. Centre national de la recherche scientifique. Institut de Chimie Radicalaire; Francia
Fil: Santos, Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biociencias, Biotecnología y Biología Traslacional; Argentina
Fil: Costantini, Paola. Università di Padova; Italia
Fil: Bortolus, Marco. Università di Padova; Italia
Fil: Carbonera, Donatella. Università di Padova; Italia
Materia
CD
EPR
FE-S CLUSTER ASSEMBLY MACHINERY
FLUORESCENCE
FRATAXIN
IRON
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/174644
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/174644
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Effects of fe2+/fe3+ binding to human frataxin and its d122y variant, as revealed by site-directed spin labeling (Sdsl) epr complemented by fluorescence and circular dichroism spectroscopiesDoni, DavidePasserini, LeonardoAudran, GérardMarque, Sylvain R. A.Schulz, MarvinSantos, JavierCostantini, PaolaBortolus, MarcoCarbonera, DonatellaCDEPRFE-S CLUSTER ASSEMBLY MACHINERYFLUORESCENCEFRATAXINIRONhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Frataxin is a highly conserved protein whose deficiency results in the neurodegenerative disease Friederich’s ataxia. Frataxin’s actual physiological function has been debated for a long time without reaching a general agreement; however, it is commonly accepted that the protein is involved in the biosynthetic iron-sulphur cluster (ISC) machinery, and several authors have pointed out that it also participates in iron homeostasis. In this work, we use site-directed spin labeling coupled to electron paramagnetic resonance (SDSL EPR) to add new information on the effects of ferric and ferrous iron binding on the properties of human frataxin in vitro. Using SDSL EPR and relating the results to fluorescence experiments commonly performed to study iron binding to FXN, we produced evidence that ferric iron causes reversible aggregation without preferred interfaces in a concentration-dependent fashion, starting at relatively low concentrations (micromolar range), whereas ferrous iron binds without inducing aggregation. Moreover, our experiments show that the ferrous binding does not lead to changes of protein conformation. The data reported in this study reveal that the currently reported binding stoichiometries should be taken with caution. The use of a spin label resistant to reduction, as well as the comparison of the binding effect of Fe2+ in wild type and in the pathological D122Y variant of frataxin, allowed us to characterize the Fe2+ binding properties of different protein sites and highlight the effect of the D122Y substitution on the surrounding residues. We suggest that both Fe2+ and Fe3+ might play a relevant role in the context of the proposed FXN physiological functions.Fil: Doni, Davide. Università di Padova; ItaliaFil: Passerini, Leonardo. Università di Padova; ItaliaFil: Audran, Gérard. Centre national de la recherche scientifique. Institut de Chimie Radicalaire; FranciaFil: Marque, Sylvain R. A.. Centre national de la recherche scientifique. Institut de Chimie Radicalaire; FranciaFil: Schulz, Marvin. Centre national de la recherche scientifique. Institut de Chimie Radicalaire; FranciaFil: Santos, Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biociencias, Biotecnología y Biología Traslacional; ArgentinaFil: Costantini, Paola. Università di Padova; ItaliaFil: Bortolus, Marco. Università di Padova; ItaliaFil: Carbonera, Donatella. Università di Padova; ItaliaMultidisciplinary Digital Publishing Institute2020-12info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/174644Doni, Davide; Passerini, Leonardo; Audran, Gérard; Marque, Sylvain R. A.; Schulz, Marvin; et al.; Effects of fe2+/fe3+ binding to human frataxin and its d122y variant, as revealed by site-directed spin labeling (Sdsl) epr complemented by fluorescence and circular dichroism spectroscopies; Multidisciplinary Digital Publishing Institute; International Journal of Molecular Sciences; 21; 24; 12-2020; 1-201661-65961422-0067CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.mdpi.com/1422-0067/21/24/9619info:eu-repo/semantics/altIdentifier/doi/10.3390/ijms21249619info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-15T14:23:34Zoai:ri.conicet.gov.ar:11336/174644instacron: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:34982025-10-15 14:23:35.139CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Effects of fe2+/fe3+ binding to human frataxin and its d122y variant, as revealed by site-directed spin labeling (Sdsl) epr complemented by fluorescence and circular dichroism spectroscopies
title Effects of fe2+/fe3+ binding to human frataxin and its d122y variant, as revealed by site-directed spin labeling (Sdsl) epr complemented by fluorescence and circular dichroism spectroscopies
spellingShingle Effects of fe2+/fe3+ binding to human frataxin and its d122y variant, as revealed by site-directed spin labeling (Sdsl) epr complemented by fluorescence and circular dichroism spectroscopies
Doni, Davide
CD
EPR
FE-S CLUSTER ASSEMBLY MACHINERY
FLUORESCENCE
FRATAXIN
IRON
title_short Effects of fe2+/fe3+ binding to human frataxin and its d122y variant, as revealed by site-directed spin labeling (Sdsl) epr complemented by fluorescence and circular dichroism spectroscopies
title_full Effects of fe2+/fe3+ binding to human frataxin and its d122y variant, as revealed by site-directed spin labeling (Sdsl) epr complemented by fluorescence and circular dichroism spectroscopies
title_fullStr Effects of fe2+/fe3+ binding to human frataxin and its d122y variant, as revealed by site-directed spin labeling (Sdsl) epr complemented by fluorescence and circular dichroism spectroscopies
title_full_unstemmed Effects of fe2+/fe3+ binding to human frataxin and its d122y variant, as revealed by site-directed spin labeling (Sdsl) epr complemented by fluorescence and circular dichroism spectroscopies
title_sort Effects of fe2+/fe3+ binding to human frataxin and its d122y variant, as revealed by site-directed spin labeling (Sdsl) epr complemented by fluorescence and circular dichroism spectroscopies
dc.creator.none.fl_str_mv Doni, Davide
Passerini, Leonardo
Audran, Gérard
Marque, Sylvain R. A.
Schulz, Marvin
Santos, Javier
Costantini, Paola
Bortolus, Marco
Carbonera, Donatella
author Doni, Davide
author_facet Doni, Davide
Passerini, Leonardo
Audran, Gérard
Marque, Sylvain R. A.
Schulz, Marvin
Santos, Javier
Costantini, Paola
Bortolus, Marco
Carbonera, Donatella
author_role author
author2 Passerini, Leonardo
Audran, Gérard
Marque, Sylvain R. A.
Schulz, Marvin
Santos, Javier
Costantini, Paola
Bortolus, Marco
Carbonera, Donatella
author2_role author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv CD
EPR
FE-S CLUSTER ASSEMBLY MACHINERY
FLUORESCENCE
FRATAXIN
IRON
topic CD
EPR
FE-S CLUSTER ASSEMBLY MACHINERY
FLUORESCENCE
FRATAXIN
IRON
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Frataxin is a highly conserved protein whose deficiency results in the neurodegenerative disease Friederich’s ataxia. Frataxin’s actual physiological function has been debated for a long time without reaching a general agreement; however, it is commonly accepted that the protein is involved in the biosynthetic iron-sulphur cluster (ISC) machinery, and several authors have pointed out that it also participates in iron homeostasis. In this work, we use site-directed spin labeling coupled to electron paramagnetic resonance (SDSL EPR) to add new information on the effects of ferric and ferrous iron binding on the properties of human frataxin in vitro. Using SDSL EPR and relating the results to fluorescence experiments commonly performed to study iron binding to FXN, we produced evidence that ferric iron causes reversible aggregation without preferred interfaces in a concentration-dependent fashion, starting at relatively low concentrations (micromolar range), whereas ferrous iron binds without inducing aggregation. Moreover, our experiments show that the ferrous binding does not lead to changes of protein conformation. The data reported in this study reveal that the currently reported binding stoichiometries should be taken with caution. The use of a spin label resistant to reduction, as well as the comparison of the binding effect of Fe2+ in wild type and in the pathological D122Y variant of frataxin, allowed us to characterize the Fe2+ binding properties of different protein sites and highlight the effect of the D122Y substitution on the surrounding residues. We suggest that both Fe2+ and Fe3+ might play a relevant role in the context of the proposed FXN physiological functions.
Fil: Doni, Davide. Università di Padova; Italia
Fil: Passerini, Leonardo. Università di Padova; Italia
Fil: Audran, Gérard. Centre national de la recherche scientifique. Institut de Chimie Radicalaire; Francia
Fil: Marque, Sylvain R. A.. Centre national de la recherche scientifique. Institut de Chimie Radicalaire; Francia
Fil: Schulz, Marvin. Centre national de la recherche scientifique. Institut de Chimie Radicalaire; Francia
Fil: Santos, Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biociencias, Biotecnología y Biología Traslacional; Argentina
Fil: Costantini, Paola. Università di Padova; Italia
Fil: Bortolus, Marco. Università di Padova; Italia
Fil: Carbonera, Donatella. Università di Padova; Italia
description Frataxin is a highly conserved protein whose deficiency results in the neurodegenerative disease Friederich’s ataxia. Frataxin’s actual physiological function has been debated for a long time without reaching a general agreement; however, it is commonly accepted that the protein is involved in the biosynthetic iron-sulphur cluster (ISC) machinery, and several authors have pointed out that it also participates in iron homeostasis. In this work, we use site-directed spin labeling coupled to electron paramagnetic resonance (SDSL EPR) to add new information on the effects of ferric and ferrous iron binding on the properties of human frataxin in vitro. Using SDSL EPR and relating the results to fluorescence experiments commonly performed to study iron binding to FXN, we produced evidence that ferric iron causes reversible aggregation without preferred interfaces in a concentration-dependent fashion, starting at relatively low concentrations (micromolar range), whereas ferrous iron binds without inducing aggregation. Moreover, our experiments show that the ferrous binding does not lead to changes of protein conformation. The data reported in this study reveal that the currently reported binding stoichiometries should be taken with caution. The use of a spin label resistant to reduction, as well as the comparison of the binding effect of Fe2+ in wild type and in the pathological D122Y variant of frataxin, allowed us to characterize the Fe2+ binding properties of different protein sites and highlight the effect of the D122Y substitution on the surrounding residues. We suggest that both Fe2+ and Fe3+ might play a relevant role in the context of the proposed FXN physiological functions.
publishDate 2020
dc.date.none.fl_str_mv 2020-12
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/11336/174644
Doni, Davide; Passerini, Leonardo; Audran, Gérard; Marque, Sylvain R. A.; Schulz, Marvin; et al.; Effects of fe2+/fe3+ binding to human frataxin and its d122y variant, as revealed by site-directed spin labeling (Sdsl) epr complemented by fluorescence and circular dichroism spectroscopies; Multidisciplinary Digital Publishing Institute; International Journal of Molecular Sciences; 21; 24; 12-2020; 1-20
1661-6596
1422-0067
CONICET Digital
CONICET
url http://hdl.handle.net/11336/174644
identifier_str_mv Doni, Davide; Passerini, Leonardo; Audran, Gérard; Marque, Sylvain R. A.; Schulz, Marvin; et al.; Effects of fe2+/fe3+ binding to human frataxin and its d122y variant, as revealed by site-directed spin labeling (Sdsl) epr complemented by fluorescence and circular dichroism spectroscopies; Multidisciplinary Digital Publishing Institute; International Journal of Molecular Sciences; 21; 24; 12-2020; 1-20
1661-6596
1422-0067
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.mdpi.com/1422-0067/21/24/9619
info:eu-repo/semantics/altIdentifier/doi/10.3390/ijms21249619
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Multidisciplinary Digital Publishing Institute
publisher.none.fl_str_mv Multidisciplinary Digital Publishing Institute
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 13.22299

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