Time-Resolved NMR Analysis of Proteolytic α-Synuclein Processing in vitro and in cellulo

Autores
Limatola, Antonio; Eichmann, Cédric; Jacob, Reeba Susan; Ben-Nissan, Gili; Sharon, Michal; Binolfi, Andrés; Selenko, Philipp
Año de publicación
2018
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Targeted proteolysis of the disordered Parkinson's disease protein alpha-synuclein (αSyn) constitutes an important event under physiological and pathological cell conditions. In this work, site-specific αSyn cleavage by different endopeptidases in vitro and by endogenous proteases in extracts of challenged and unchallenged cells was studied by time-resolved NMR spectroscopy. Specifically, proteolytic processing was monitored under neutral and low pH conditions and in response to Rotenone-induced oxidative stress. Further, time-dependent degradation of electroporation-delivered αSyn in intact SH-SY5Y and A2780 cells was analyzed. Results presented here delineate a general framework for NMR-based proteolysis studies in vitro and in cellulo, and confirm earlier reports pertaining to the exceptional proteolytic stability of αSyn under physiological cell conditions. However, experimental findings also reveal altered protease susceptibilities in selected mammalian cell lines and upon induced cell stress.
Fil: Limatola, Antonio. Leibniz Institute Of Molecular Pharmacology; Alemania
Fil: Eichmann, Cédric. Leibniz Institute Of Molecular Pharmacology; Alemania
Fil: Jacob, Reeba Susan. Leibniz Institute Of Molecular Pharmacology; Alemania
Fil: Ben-Nissan, Gili. No especifíca;
Fil: Sharon, Michal. No especifíca;
Fil: Binolfi, Andrés. 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: Selenko, Philipp. No especifíca;
Materia
ALPHA-SYNUCLEIN
IN-CELL NMR
PARKINSON'S DISEASE
POST-TRANSLATIONAL MODIFICATIONS
PROTEOLYTIC PROCESSING
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:11336/185916
Acceder
id CONICETDig_2d3e1a5fd29816d2a5847b9427041a88
oai_identifier_str oai:ri.conicet.gov.ar:11336/185916
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Time-Resolved NMR Analysis of Proteolytic α-Synuclein Processing in vitro and in celluloLimatola, AntonioEichmann, CédricJacob, Reeba SusanBen-Nissan, GiliSharon, MichalBinolfi, AndrésSelenko, PhilippALPHA-SYNUCLEININ-CELL NMRPARKINSON'S DISEASEPOST-TRANSLATIONAL MODIFICATIONSPROTEOLYTIC PROCESSINGhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Targeted proteolysis of the disordered Parkinson's disease protein alpha-synuclein (αSyn) constitutes an important event under physiological and pathological cell conditions. In this work, site-specific αSyn cleavage by different endopeptidases in vitro and by endogenous proteases in extracts of challenged and unchallenged cells was studied by time-resolved NMR spectroscopy. Specifically, proteolytic processing was monitored under neutral and low pH conditions and in response to Rotenone-induced oxidative stress. Further, time-dependent degradation of electroporation-delivered αSyn in intact SH-SY5Y and A2780 cells was analyzed. Results presented here delineate a general framework for NMR-based proteolysis studies in vitro and in cellulo, and confirm earlier reports pertaining to the exceptional proteolytic stability of αSyn under physiological cell conditions. However, experimental findings also reveal altered protease susceptibilities in selected mammalian cell lines and upon induced cell stress.Fil: Limatola, Antonio. Leibniz Institute Of Molecular Pharmacology; AlemaniaFil: Eichmann, Cédric. Leibniz Institute Of Molecular Pharmacology; AlemaniaFil: Jacob, Reeba Susan. Leibniz Institute Of Molecular Pharmacology; AlemaniaFil: Ben-Nissan, Gili. No especifíca;Fil: Sharon, Michal. No especifíca;Fil: Binolfi, Andrés. 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: Selenko, Philipp. No especifíca;Wiley VCH Verlag2018-11info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/185916Limatola, Antonio; Eichmann, Cédric; Jacob, Reeba Susan; Ben-Nissan, Gili; Sharon, Michal; et al.; Time-Resolved NMR Analysis of Proteolytic α-Synuclein Processing in vitro and in cellulo; Wiley VCH Verlag; Proteomics (weinheim. Print); 18; 21-22; 11-2018; 1-131615-9853CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/http://doi.wiley.com/10.1002/pmic.201800056info:eu-repo/semantics/altIdentifier/doi/10.1002/pmic.201800056info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T09:44:52Zoai:ri.conicet.gov.ar:11336/185916instacron: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:34982025-09-03 09:44:52.917CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Time-Resolved NMR Analysis of Proteolytic α-Synuclein Processing in vitro and in cellulo
title Time-Resolved NMR Analysis of Proteolytic α-Synuclein Processing in vitro and in cellulo
spellingShingle Time-Resolved NMR Analysis of Proteolytic α-Synuclein Processing in vitro and in cellulo
Limatola, Antonio
ALPHA-SYNUCLEIN
IN-CELL NMR
PARKINSON'S DISEASE
POST-TRANSLATIONAL MODIFICATIONS
PROTEOLYTIC PROCESSING
title_short Time-Resolved NMR Analysis of Proteolytic α-Synuclein Processing in vitro and in cellulo
title_full Time-Resolved NMR Analysis of Proteolytic α-Synuclein Processing in vitro and in cellulo
title_fullStr Time-Resolved NMR Analysis of Proteolytic α-Synuclein Processing in vitro and in cellulo
title_full_unstemmed Time-Resolved NMR Analysis of Proteolytic α-Synuclein Processing in vitro and in cellulo
title_sort Time-Resolved NMR Analysis of Proteolytic α-Synuclein Processing in vitro and in cellulo
dc.creator.none.fl_str_mv Limatola, Antonio
Eichmann, Cédric
Jacob, Reeba Susan
Ben-Nissan, Gili
Sharon, Michal
Binolfi, Andrés
Selenko, Philipp
author Limatola, Antonio
author_facet Limatola, Antonio
Eichmann, Cédric
Jacob, Reeba Susan
Ben-Nissan, Gili
Sharon, Michal
Binolfi, Andrés
Selenko, Philipp
author_role author
author2 Eichmann, Cédric
Jacob, Reeba Susan
Ben-Nissan, Gili
Sharon, Michal
Binolfi, Andrés
Selenko, Philipp
author2_role author
author
author
author
author
author
dc.subject.none.fl_str_mv ALPHA-SYNUCLEIN
IN-CELL NMR
PARKINSON'S DISEASE
POST-TRANSLATIONAL MODIFICATIONS
PROTEOLYTIC PROCESSING
topic ALPHA-SYNUCLEIN
IN-CELL NMR
PARKINSON'S DISEASE
POST-TRANSLATIONAL MODIFICATIONS
PROTEOLYTIC PROCESSING
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Targeted proteolysis of the disordered Parkinson's disease protein alpha-synuclein (αSyn) constitutes an important event under physiological and pathological cell conditions. In this work, site-specific αSyn cleavage by different endopeptidases in vitro and by endogenous proteases in extracts of challenged and unchallenged cells was studied by time-resolved NMR spectroscopy. Specifically, proteolytic processing was monitored under neutral and low pH conditions and in response to Rotenone-induced oxidative stress. Further, time-dependent degradation of electroporation-delivered αSyn in intact SH-SY5Y and A2780 cells was analyzed. Results presented here delineate a general framework for NMR-based proteolysis studies in vitro and in cellulo, and confirm earlier reports pertaining to the exceptional proteolytic stability of αSyn under physiological cell conditions. However, experimental findings also reveal altered protease susceptibilities in selected mammalian cell lines and upon induced cell stress.
Fil: Limatola, Antonio. Leibniz Institute Of Molecular Pharmacology; Alemania
Fil: Eichmann, Cédric. Leibniz Institute Of Molecular Pharmacology; Alemania
Fil: Jacob, Reeba Susan. Leibniz Institute Of Molecular Pharmacology; Alemania
Fil: Ben-Nissan, Gili. No especifíca;
Fil: Sharon, Michal. No especifíca;
Fil: Binolfi, Andrés. 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: Selenko, Philipp. No especifíca;
description Targeted proteolysis of the disordered Parkinson's disease protein alpha-synuclein (αSyn) constitutes an important event under physiological and pathological cell conditions. In this work, site-specific αSyn cleavage by different endopeptidases in vitro and by endogenous proteases in extracts of challenged and unchallenged cells was studied by time-resolved NMR spectroscopy. Specifically, proteolytic processing was monitored under neutral and low pH conditions and in response to Rotenone-induced oxidative stress. Further, time-dependent degradation of electroporation-delivered αSyn in intact SH-SY5Y and A2780 cells was analyzed. Results presented here delineate a general framework for NMR-based proteolysis studies in vitro and in cellulo, and confirm earlier reports pertaining to the exceptional proteolytic stability of αSyn under physiological cell conditions. However, experimental findings also reveal altered protease susceptibilities in selected mammalian cell lines and upon induced cell stress.
publishDate 2018
dc.date.none.fl_str_mv 2018-11
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/11336/185916
Limatola, Antonio; Eichmann, Cédric; Jacob, Reeba Susan; Ben-Nissan, Gili; Sharon, Michal; et al.; Time-Resolved NMR Analysis of Proteolytic α-Synuclein Processing in vitro and in cellulo; Wiley VCH Verlag; Proteomics (weinheim. Print); 18; 21-22; 11-2018; 1-13
1615-9853
CONICET Digital
CONICET
url http://hdl.handle.net/11336/185916
identifier_str_mv Limatola, Antonio; Eichmann, Cédric; Jacob, Reeba Susan; Ben-Nissan, Gili; Sharon, Michal; et al.; Time-Resolved NMR Analysis of Proteolytic α-Synuclein Processing in vitro and in cellulo; Wiley VCH Verlag; Proteomics (weinheim. Print); 18; 21-22; 11-2018; 1-13
1615-9853
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/http://doi.wiley.com/10.1002/pmic.201800056
info:eu-repo/semantics/altIdentifier/doi/10.1002/pmic.201800056
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 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_ 1842268694700359680
score 13.13397

Publicaciones similares