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
- Institución
- Universidad Nacional de Buenos Aires. Facultad de Ciencias Exactas y Naturales
- OAI Identificador
- paperaa:paper_19326203_v7_n9_p_Roman
Ver los metadatos del registro completo
id |
BDUBAFCEN_14ccdd642ff673a184ad220b4a9d5e9b |
---|---|
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 |
_version_ |
1844618734103166976 |
score |
13.070432 |