Direct Measurement of the Nanomechanical Stability of a Redox Protein Active Site and Its Dependence upon Metal Binding

The structural basis of the low reorganization energy of cupredoxins has long been debated. These proteins reconcile a conformationally heterogeneous and exposed metal-chelating site with the highly rigid copper center required for efficient electron transfer. Here we combine single-molecule mechani...

ver descrição completa

Detalhes bibliográficos
Autores: Giannotti, Marina, Cabeza de Vaca, Israel, Artés, Juan M., Sanz, Fausto, Guallar, Víctor|||0000-0002-4580-1114, Gorostiza, Pau
Tipo de documento: artigo
Data de publicação:2015
País:España
Recursos:Universitat Politècnica de Catalunya (UPC)
Repositório:UPCommons. Portal del coneixement obert de la UPC
Idioma:inglês
OAI Identifier:oai:upcommons.upc.edu:2117/84165
Acesso em linha:https://hdl.handle.net/2117/84165
https://dx.doi.org/10.1021/acs.jpcb.5b06382
Access Level:Acceso aberto
Palavra-chave:Protein
Azurin
Cupredoxins
Force spectroscopy
Nanomechanical stability
Single molecule
Proteïnes
Àrees temàtiques de la UPC::Enginyeria mecànica::Impacte ambiental
id ES_acb65c5a5017fc659e1be0a8bc2b21fb
oai_identifier_str oai:upcommons.upc.edu:2117/84165
network_acronym_str ES
network_name_str España
repository_id_str
spelling Direct Measurement of the Nanomechanical Stability of a Redox Protein Active Site and Its Dependence upon Metal BindingGiannotti, MarinaCabeza de Vaca, IsraelArtés, Juan M.Sanz, FaustoGuallar, Víctor|||0000-0002-4580-1114Gorostiza, PauProteinAzurinCupredoxinsForce spectroscopyNanomechanical stabilitySingle moleculeProteïnesÀrees temàtiques de la UPC::Enginyeria mecànica::Impacte ambientalThe structural basis of the low reorganization energy of cupredoxins has long been debated. These proteins reconcile a conformationally heterogeneous and exposed metal-chelating site with the highly rigid copper center required for efficient electron transfer. Here we combine single-molecule mechanical unfolding experiments with statistical analysis and computer simulations to show that the metal-binding region of apo-azurin is mechanically flexible and that high mechanical stability is imparted by copper binding. The unfolding pathway of the metal site depends on the pulling residue and suggests that partial unfolding of the metal binding site could be facilitated by the physical interaction with certain regions of the redox protein.We are grateful to A. Donaire and I. Díez-Pérez for discussions, and to the Catalan government (grant 2014SGR-1251), the Spanish government (grant CTQ2013-43892R) and the European Research Council (PELE ERC-2009-Adg 25027) for financial support.Peer ReviewedACS Publications20152015-09-1020162016-03-10journal articlehttp://purl.org/coar/resource_type/c_6501AMhttp://purl.org/coar/version/c_ab4af688f83e57aainfo:eu-repo/semantics/articleapplication/pdfhttps://hdl.handle.net/2117/84165https://dx.doi.org/10.1021/acs.jpcb.5b06382reponame:UPCommons. Portal del coneixement obert de la UPCinstname:Universitat Politècnica de Catalunya (UPC)InglésengEuropean Commission http://dx.doi.org/10.13039/100011102 Seventh Framework Programme 250277 P.E.L.E (Protein Energy Landscape Exploration): a la carte drug design toolsMinisterio de Economía y Competitividad http://doi.org/10.13039/501100003329 CTQ2013-43892-R APLICACIONES TERAPEUTICAS DE LA OPTOFARMACOLOGIAopen accesshttp://purl.org/coar/access_right/c_abf2info:eu-repo/semantics/openAccessoai:upcommons.upc.edu:2117/841652026-05-27T15:37:01Z
dc.title.none.fl_str_mv Direct Measurement of the Nanomechanical Stability of a Redox Protein Active Site and Its Dependence upon Metal Binding
title Direct Measurement of the Nanomechanical Stability of a Redox Protein Active Site and Its Dependence upon Metal Binding
spellingShingle Direct Measurement of the Nanomechanical Stability of a Redox Protein Active Site and Its Dependence upon Metal Binding
Giannotti, Marina
Protein
Azurin
Cupredoxins
Force spectroscopy
Nanomechanical stability
Single molecule
Proteïnes
Àrees temàtiques de la UPC::Enginyeria mecànica::Impacte ambiental
title_short Direct Measurement of the Nanomechanical Stability of a Redox Protein Active Site and Its Dependence upon Metal Binding
title_full Direct Measurement of the Nanomechanical Stability of a Redox Protein Active Site and Its Dependence upon Metal Binding
title_fullStr Direct Measurement of the Nanomechanical Stability of a Redox Protein Active Site and Its Dependence upon Metal Binding
title_full_unstemmed Direct Measurement of the Nanomechanical Stability of a Redox Protein Active Site and Its Dependence upon Metal Binding
title_sort Direct Measurement of the Nanomechanical Stability of a Redox Protein Active Site and Its Dependence upon Metal Binding
dc.creator.none.fl_str_mv Giannotti, Marina
Cabeza de Vaca, Israel
Artés, Juan M.
Sanz, Fausto
Guallar, Víctor|||0000-0002-4580-1114
Gorostiza, Pau
author Giannotti, Marina
author_facet Giannotti, Marina
Cabeza de Vaca, Israel
Artés, Juan M.
Sanz, Fausto
Guallar, Víctor|||0000-0002-4580-1114
Gorostiza, Pau
author_role author
author2 Cabeza de Vaca, Israel
Artés, Juan M.
Sanz, Fausto
Guallar, Víctor|||0000-0002-4580-1114
Gorostiza, Pau
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Protein
Azurin
Cupredoxins
Force spectroscopy
Nanomechanical stability
Single molecule
Proteïnes
Àrees temàtiques de la UPC::Enginyeria mecànica::Impacte ambiental
topic Protein
Azurin
Cupredoxins
Force spectroscopy
Nanomechanical stability
Single molecule
Proteïnes
Àrees temàtiques de la UPC::Enginyeria mecànica::Impacte ambiental
description The structural basis of the low reorganization energy of cupredoxins has long been debated. These proteins reconcile a conformationally heterogeneous and exposed metal-chelating site with the highly rigid copper center required for efficient electron transfer. Here we combine single-molecule mechanical unfolding experiments with statistical analysis and computer simulations to show that the metal-binding region of apo-azurin is mechanically flexible and that high mechanical stability is imparted by copper binding. The unfolding pathway of the metal site depends on the pulling residue and suggests that partial unfolding of the metal binding site could be facilitated by the physical interaction with certain regions of the redox protein.
publishDate 2015
dc.date.none.fl_str_mv 2015
2015-09-10
2016
2016-03-10
dc.type.none.fl_str_mv journal article
http://purl.org/coar/resource_type/c_6501
AM
http://purl.org/coar/version/c_ab4af688f83e57aa
dc.type.openaire.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv https://hdl.handle.net/2117/84165
https://dx.doi.org/10.1021/acs.jpcb.5b06382
url https://hdl.handle.net/2117/84165
https://dx.doi.org/10.1021/acs.jpcb.5b06382
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.relation.none.fl_str_mv European Commission http://dx.doi.org/10.13039/100011102 Seventh Framework Programme 250277 P.E.L.E (Protein Energy Landscape Exploration): a la carte drug design tools
Ministerio de Economía y Competitividad http://doi.org/10.13039/501100003329 CTQ2013-43892-R APLICACIONES TERAPEUTICAS DE LA OPTOFARMACOLOGIA
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
dc.rights.openaire.fl_str_mv info:eu-repo/semantics/openAccess
rights_invalid_str_mv open access
http://purl.org/coar/access_right/c_abf2
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv ACS Publications
publisher.none.fl_str_mv ACS Publications
dc.source.none.fl_str_mv reponame:UPCommons. Portal del coneixement obert de la UPC
instname:Universitat Politècnica de Catalunya (UPC)
instname_str Universitat Politècnica de Catalunya (UPC)
reponame_str UPCommons. Portal del coneixement obert de la UPC
collection UPCommons. Portal del coneixement obert de la UPC
repository.name.fl_str_mv
repository.mail.fl_str_mv
_version_ 1869416378369310720
score 15.301603

Registros relacionados