Protein Stability and Dynamics Modulation: The Case of Human Frataxin

Autores
Roman, E.A.; Faraj, S.E.; Gallo, M.; Salvay, A.G.; Ferreiro, D.U.; Santos, J.
Año de publicación
2012
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Frataxin (FXN) is an α/β protein that plays an essential role in iron homeostasis. Apparently, the function of human FXN (hFXN) depends on the cooperative formation of crucial interactions between helix α1, helix α2, and the C-terminal region (CTR) of the protein. In this work we quantitatively explore these relationships using a purified recombinant fragment hFXN90-195. This variant shows the hydrodynamic behavior expected for a monomeric globular domain. Circular dichroism, fluorescence, and NMR spectroscopies show that hFXN90-195 presents native-like secondary and tertiary structure. However, chemical and temperature induced denaturation show that CTR truncation significantly destabilizes the overall hFXN fold. Accordingly, limited proteolysis experiments suggest that the native-state dynamics of hFXN90-195 and hFXN90-210 are indeed different, being the former form much more sensitive to the protease at specific sites. The overall folding dynamics of hFXN fold was further explored with structure-based protein folding simulations. These suggest that the native ensemble of hFXN can be decomposed in at least two substates, one with consolidation of the CTR and the other without consolidation of the CTR. Explicit-solvent all atom simulations identify some of the proteolytic target sites as flexible regions of the protein. We propose that the local unfolding of CTR may be a critical step for the global unfolding of hFXN, and that modulation of the CTR interactions may strongly affect hFXN physiological function. © 2012 Roman et al.
Fil:Roman, E.A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.
Fil:Gallo, M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.
Fil:Ferreiro, D.U. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.
Fuente
PLoS ONE 2012;7(9)
Materia
frataxin
alpha helix
article
carboxy terminal sequence
circular dichroism
fluorescence analysis
hydrodynamics
nuclear magnetic resonance spectroscopy
protein folding
protein function
protein secondary structure
protein stability
protein tertiary structure
protein unfolding
quantitative analysis
structure analysis
Circular Dichroism
Homeostasis
Humans
Hydrodynamics
Iron
Iron-Binding Proteins
Magnetic Resonance Spectroscopy
Microscopy, Fluorescence
Models, Molecular
Molecular Conformation
Molecular Dynamics Simulation
Point Mutation
Protein Denaturation
Protein Folding
Protein Structure, Tertiary
Recombinant Proteins
Solvents
Temperature
Time Factors
Nivel de accesibilidad
acceso abierto
Condiciones de uso
http://creativecommons.org/licenses/by/2.5/ar
Repositorio
Biblioteca Digital (UBA-FCEN)
Institución
Universidad Nacional de Buenos Aires. Facultad de Ciencias Exactas y Naturales
OAI Identificador
paperaa:paper_19326203_v7_n9_p_Roman

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oai_identifier_str paperaa:paper_19326203_v7_n9_p_Roman
network_acronym_str BDUBAFCEN
repository_id_str 1896
network_name_str Biblioteca Digital (UBA-FCEN)
spelling Protein Stability and Dynamics Modulation: The Case of Human FrataxinRoman, E.A.Faraj, S.E.Gallo, M.Salvay, A.G.Ferreiro, D.U.Santos, J.frataxinalpha helixarticlecarboxy terminal sequencecircular dichroismfluorescence analysishydrodynamicsnuclear magnetic resonance spectroscopyprotein foldingprotein functionprotein secondary structureprotein stabilityprotein tertiary structureprotein unfoldingquantitative analysisstructure analysisCircular DichroismHomeostasisHumansHydrodynamicsIronIron-Binding ProteinsMagnetic Resonance SpectroscopyMicroscopy, FluorescenceModels, MolecularMolecular ConformationMolecular Dynamics SimulationPoint MutationProtein DenaturationProtein FoldingProtein Structure, TertiaryRecombinant ProteinsSolventsTemperatureTime FactorsFrataxin (FXN) is an α/β protein that plays an essential role in iron homeostasis. Apparently, the function of human FXN (hFXN) depends on the cooperative formation of crucial interactions between helix α1, helix α2, and the C-terminal region (CTR) of the protein. In this work we quantitatively explore these relationships using a purified recombinant fragment hFXN90-195. This variant shows the hydrodynamic behavior expected for a monomeric globular domain. Circular dichroism, fluorescence, and NMR spectroscopies show that hFXN90-195 presents native-like secondary and tertiary structure. However, chemical and temperature induced denaturation show that CTR truncation significantly destabilizes the overall hFXN fold. Accordingly, limited proteolysis experiments suggest that the native-state dynamics of hFXN90-195 and hFXN90-210 are indeed different, being the former form much more sensitive to the protease at specific sites. The overall folding dynamics of hFXN fold was further explored with structure-based protein folding simulations. These suggest that the native ensemble of hFXN can be decomposed in at least two substates, one with consolidation of the CTR and the other without consolidation of the CTR. Explicit-solvent all atom simulations identify some of the proteolytic target sites as flexible regions of the protein. We propose that the local unfolding of CTR may be a critical step for the global unfolding of hFXN, and that modulation of the CTR interactions may strongly affect hFXN physiological function. © 2012 Roman et al.Fil:Roman, E.A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.Fil:Gallo, M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.Fil:Ferreiro, D.U. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.2012info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttp://hdl.handle.net/20.500.12110/paper_19326203_v7_n9_p_RomanPLoS ONE 2012;7(9)reponame:Biblioteca Digital (UBA-FCEN)instname:Universidad Nacional de Buenos Aires. Facultad de Ciencias Exactas y Naturalesinstacron:UBA-FCENenginfo:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by/2.5/ar2025-09-29T13:42:52Zpaperaa:paper_19326203_v7_n9_p_RomanInstitucionalhttps://digital.bl.fcen.uba.ar/Universidad públicaNo correspondehttps://digital.bl.fcen.uba.ar/cgi-bin/oaiserver.cgiana@bl.fcen.uba.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:18962025-09-29 13:42:53.396Biblioteca Digital (UBA-FCEN) - Universidad Nacional de Buenos Aires. Facultad de Ciencias Exactas y Naturalesfalse
dc.title.none.fl_str_mv Protein Stability and Dynamics Modulation: The Case of Human Frataxin
title Protein Stability and Dynamics Modulation: The Case of Human Frataxin
spellingShingle Protein Stability and Dynamics Modulation: The Case of Human Frataxin
Roman, E.A.
frataxin
alpha helix
article
carboxy terminal sequence
circular dichroism
fluorescence analysis
hydrodynamics
nuclear magnetic resonance spectroscopy
protein folding
protein function
protein secondary structure
protein stability
protein tertiary structure
protein unfolding
quantitative analysis
structure analysis
Circular Dichroism
Homeostasis
Humans
Hydrodynamics
Iron
Iron-Binding Proteins
Magnetic Resonance Spectroscopy
Microscopy, Fluorescence
Models, Molecular
Molecular Conformation
Molecular Dynamics Simulation
Point Mutation
Protein Denaturation
Protein Folding
Protein Structure, Tertiary
Recombinant Proteins
Solvents
Temperature
Time Factors
title_short Protein Stability and Dynamics Modulation: The Case of Human Frataxin
title_full Protein Stability and Dynamics Modulation: The Case of Human Frataxin
title_fullStr Protein Stability and Dynamics Modulation: The Case of Human Frataxin
title_full_unstemmed Protein Stability and Dynamics Modulation: The Case of Human Frataxin
title_sort Protein Stability and Dynamics Modulation: The Case of Human Frataxin
dc.creator.none.fl_str_mv Roman, E.A.
Faraj, S.E.
Gallo, M.
Salvay, A.G.
Ferreiro, D.U.
Santos, J.
author Roman, E.A.
author_facet Roman, E.A.
Faraj, S.E.
Gallo, M.
Salvay, A.G.
Ferreiro, D.U.
Santos, J.
author_role author
author2 Faraj, S.E.
Gallo, M.
Salvay, A.G.
Ferreiro, D.U.
Santos, J.
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv frataxin
alpha helix
article
carboxy terminal sequence
circular dichroism
fluorescence analysis
hydrodynamics
nuclear magnetic resonance spectroscopy
protein folding
protein function
protein secondary structure
protein stability
protein tertiary structure
protein unfolding
quantitative analysis
structure analysis
Circular Dichroism
Homeostasis
Humans
Hydrodynamics
Iron
Iron-Binding Proteins
Magnetic Resonance Spectroscopy
Microscopy, Fluorescence
Models, Molecular
Molecular Conformation
Molecular Dynamics Simulation
Point Mutation
Protein Denaturation
Protein Folding
Protein Structure, Tertiary
Recombinant Proteins
Solvents
Temperature
Time Factors
topic frataxin
alpha helix
article
carboxy terminal sequence
circular dichroism
fluorescence analysis
hydrodynamics
nuclear magnetic resonance spectroscopy
protein folding
protein function
protein secondary structure
protein stability
protein tertiary structure
protein unfolding
quantitative analysis
structure analysis
Circular Dichroism
Homeostasis
Humans
Hydrodynamics
Iron
Iron-Binding Proteins
Magnetic Resonance Spectroscopy
Microscopy, Fluorescence
Models, Molecular
Molecular Conformation
Molecular Dynamics Simulation
Point Mutation
Protein Denaturation
Protein Folding
Protein Structure, Tertiary
Recombinant Proteins
Solvents
Temperature
Time Factors
dc.description.none.fl_txt_mv Frataxin (FXN) is an α/β protein that plays an essential role in iron homeostasis. Apparently, the function of human FXN (hFXN) depends on the cooperative formation of crucial interactions between helix α1, helix α2, and the C-terminal region (CTR) of the protein. In this work we quantitatively explore these relationships using a purified recombinant fragment hFXN90-195. This variant shows the hydrodynamic behavior expected for a monomeric globular domain. Circular dichroism, fluorescence, and NMR spectroscopies show that hFXN90-195 presents native-like secondary and tertiary structure. However, chemical and temperature induced denaturation show that CTR truncation significantly destabilizes the overall hFXN fold. Accordingly, limited proteolysis experiments suggest that the native-state dynamics of hFXN90-195 and hFXN90-210 are indeed different, being the former form much more sensitive to the protease at specific sites. The overall folding dynamics of hFXN fold was further explored with structure-based protein folding simulations. These suggest that the native ensemble of hFXN can be decomposed in at least two substates, one with consolidation of the CTR and the other without consolidation of the CTR. Explicit-solvent all atom simulations identify some of the proteolytic target sites as flexible regions of the protein. We propose that the local unfolding of CTR may be a critical step for the global unfolding of hFXN, and that modulation of the CTR interactions may strongly affect hFXN physiological function. © 2012 Roman et al.
Fil:Roman, E.A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.
Fil:Gallo, M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.
Fil:Ferreiro, D.U. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.
description Frataxin (FXN) is an α/β protein that plays an essential role in iron homeostasis. Apparently, the function of human FXN (hFXN) depends on the cooperative formation of crucial interactions between helix α1, helix α2, and the C-terminal region (CTR) of the protein. In this work we quantitatively explore these relationships using a purified recombinant fragment hFXN90-195. This variant shows the hydrodynamic behavior expected for a monomeric globular domain. Circular dichroism, fluorescence, and NMR spectroscopies show that hFXN90-195 presents native-like secondary and tertiary structure. However, chemical and temperature induced denaturation show that CTR truncation significantly destabilizes the overall hFXN fold. Accordingly, limited proteolysis experiments suggest that the native-state dynamics of hFXN90-195 and hFXN90-210 are indeed different, being the former form much more sensitive to the protease at specific sites. The overall folding dynamics of hFXN fold was further explored with structure-based protein folding simulations. These suggest that the native ensemble of hFXN can be decomposed in at least two substates, one with consolidation of the CTR and the other without consolidation of the CTR. Explicit-solvent all atom simulations identify some of the proteolytic target sites as flexible regions of the protein. We propose that the local unfolding of CTR may be a critical step for the global unfolding of hFXN, and that modulation of the CTR interactions may strongly affect hFXN physiological function. © 2012 Roman et al.
publishDate 2012
dc.date.none.fl_str_mv 2012
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/20.500.12110/paper_19326203_v7_n9_p_Roman
url http://hdl.handle.net/20.500.12110/paper_19326203_v7_n9_p_Roman
dc.language.none.fl_str_mv eng
language eng
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by/2.5/ar
eu_rights_str_mv openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by/2.5/ar
dc.format.none.fl_str_mv application/pdf
dc.source.none.fl_str_mv PLoS ONE 2012;7(9)
reponame:Biblioteca Digital (UBA-FCEN)
instname:Universidad Nacional de Buenos Aires. Facultad de Ciencias Exactas y Naturales
instacron:UBA-FCEN
reponame_str Biblioteca Digital (UBA-FCEN)
collection Biblioteca Digital (UBA-FCEN)
instname_str Universidad Nacional de Buenos Aires. Facultad de Ciencias Exactas y Naturales
instacron_str UBA-FCEN
institution UBA-FCEN
repository.name.fl_str_mv Biblioteca Digital (UBA-FCEN) - Universidad Nacional de Buenos Aires. Facultad de Ciencias Exactas y Naturales
repository.mail.fl_str_mv ana@bl.fcen.uba.ar
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