The Use of Mn(II) Bound to His-tags as Genetically Encodable Spin-Label for Nanometric Distance Determination in Proteins

Autores
Ching, H. Y. Vincent; Mascali, Florencia Carla; Bertrand, Hélène C.; Bruch, Eduardo Marcos; Demay Drouhard, Paul; Rasia, Rodolfo Maximiliano; Policar, Clotilde; Tabares, Leandro Cesar; Un, Sun
Año de publicación
2016
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
A genetically encodable paramagnetic spin-label capable of self-assembly from naturally available components would offer a means for studying the in-cell structure and interactions of a protein by electron paramagnetic resonance (EPR). Here, we demonstrate pulse electron-electron double resonance (DEER) measurements on spin-labels consisting of Mn(II) ions coordinated to a sequence of histidines, so-called His-tags, that are ubiquitously added by genetic engineering to facilitate protein purification. Although the affinity of His-tags for Mn(II) was low (800 μM), Mn(II)-bound His-tags yielded readily detectable DEER time traces even at concentrations expected in cells. We were able to determine accurately the distance between two His-tag Mn(II) spin-labels at the ends of a rigid helical polyproline peptide of known structure, as well as at the ends of a completely cell-synthesized 3-helix bundle. This approach not only greatly simplifies the labeling procedure but also represents a first step towards using self-assembling metal spin-labels for in-cell distance measurements.
Fil: Ching, H. Y. Vincent. Université Paris-Saclay; Francia
Fil: Mascali, Florencia Carla. 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: Bertrand, Hélène C.. Ecole Normale Supérieure; Francia. Centre National de la Recherche Scientifique; Francia. Universite de Paris VI; Francia
Fil: Bruch, Eduardo Marcos. Université Paris-Saclay; Francia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Demay Drouhard, Paul. Ecole Normale Supérieure; Francia. Centre National de la Recherche Scientifique; Francia. Universite de Paris VI; Francia
Fil: Rasia, Rodolfo Maximiliano. 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: Policar, Clotilde. Ecole Normale Supérieure; Francia. Centre National de la Recherche Scientifique; Francia. Universite de Paris VI; Francia
Fil: Tabares, Leandro Cesar. Université Paris-Saclay; Francia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Un, Sun. Université Paris-Saclay; Francia
Materia
Mn(Ii)
Epr
Deer
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:11336/61004
Acceder
id CONICETDig_f4be4b025c3cd75db4ecb355c173635f
oai_identifier_str oai:ri.conicet.gov.ar:11336/61004
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling The Use of Mn(II) Bound to His-tags as Genetically Encodable Spin-Label for Nanometric Distance Determination in ProteinsChing, H. Y. VincentMascali, Florencia CarlaBertrand, Hélène C.Bruch, Eduardo MarcosDemay Drouhard, PaulRasia, Rodolfo MaximilianoPolicar, ClotildeTabares, Leandro CesarUn, SunMn(Ii)EprDeerhttps://purl.org/becyt/ford/1.3https://purl.org/becyt/ford/1https://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1A genetically encodable paramagnetic spin-label capable of self-assembly from naturally available components would offer a means for studying the in-cell structure and interactions of a protein by electron paramagnetic resonance (EPR). Here, we demonstrate pulse electron-electron double resonance (DEER) measurements on spin-labels consisting of Mn(II) ions coordinated to a sequence of histidines, so-called His-tags, that are ubiquitously added by genetic engineering to facilitate protein purification. Although the affinity of His-tags for Mn(II) was low (800 μM), Mn(II)-bound His-tags yielded readily detectable DEER time traces even at concentrations expected in cells. We were able to determine accurately the distance between two His-tag Mn(II) spin-labels at the ends of a rigid helical polyproline peptide of known structure, as well as at the ends of a completely cell-synthesized 3-helix bundle. This approach not only greatly simplifies the labeling procedure but also represents a first step towards using self-assembling metal spin-labels for in-cell distance measurements.Fil: Ching, H. Y. Vincent. Université Paris-Saclay; FranciaFil: Mascali, Florencia Carla. 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: Bertrand, Hélène C.. Ecole Normale Supérieure; Francia. Centre National de la Recherche Scientifique; Francia. Universite de Paris VI; FranciaFil: Bruch, Eduardo Marcos. Université Paris-Saclay; Francia. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Demay Drouhard, Paul. Ecole Normale Supérieure; Francia. Centre National de la Recherche Scientifique; Francia. Universite de Paris VI; FranciaFil: Rasia, Rodolfo Maximiliano. 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: Policar, Clotilde. Ecole Normale Supérieure; Francia. Centre National de la Recherche Scientifique; Francia. Universite de Paris VI; FranciaFil: Tabares, Leandro Cesar. Université Paris-Saclay; Francia. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Un, Sun. Université Paris-Saclay; FranciaAmerican Chemical Society2016-03info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/61004Ching, H. Y. Vincent; Mascali, Florencia Carla; Bertrand, Hélène C.; Bruch, Eduardo Marcos; Demay Drouhard, Paul; et al.; The Use of Mn(II) Bound to His-tags as Genetically Encodable Spin-Label for Nanometric Distance Determination in Proteins; American Chemical Society; Journal of Physical Chemistry Letters; 7; 6; 3-2016; 1072-10761948-7185CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1021/acs.jpclett.6b00362info:eu-repo/semantics/altIdentifier/url/https://pubs.acs.org/doi/10.1021/acs.jpclett.6b00362info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-15T14:42:47Zoai:ri.conicet.gov.ar:11336/61004instacron: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 14:42:47.583CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv The Use of Mn(II) Bound to His-tags as Genetically Encodable Spin-Label for Nanometric Distance Determination in Proteins
title The Use of Mn(II) Bound to His-tags as Genetically Encodable Spin-Label for Nanometric Distance Determination in Proteins
spellingShingle The Use of Mn(II) Bound to His-tags as Genetically Encodable Spin-Label for Nanometric Distance Determination in Proteins
Ching, H. Y. Vincent
Mn(Ii)
Epr
Deer
title_short The Use of Mn(II) Bound to His-tags as Genetically Encodable Spin-Label for Nanometric Distance Determination in Proteins
title_full The Use of Mn(II) Bound to His-tags as Genetically Encodable Spin-Label for Nanometric Distance Determination in Proteins
title_fullStr The Use of Mn(II) Bound to His-tags as Genetically Encodable Spin-Label for Nanometric Distance Determination in Proteins
title_full_unstemmed The Use of Mn(II) Bound to His-tags as Genetically Encodable Spin-Label for Nanometric Distance Determination in Proteins
title_sort The Use of Mn(II) Bound to His-tags as Genetically Encodable Spin-Label for Nanometric Distance Determination in Proteins
dc.creator.none.fl_str_mv Ching, H. Y. Vincent
Mascali, Florencia Carla
Bertrand, Hélène C.
Bruch, Eduardo Marcos
Demay Drouhard, Paul
Rasia, Rodolfo Maximiliano
Policar, Clotilde
Tabares, Leandro Cesar
Un, Sun
author Ching, H. Y. Vincent
author_facet Ching, H. Y. Vincent
Mascali, Florencia Carla
Bertrand, Hélène C.
Bruch, Eduardo Marcos
Demay Drouhard, Paul
Rasia, Rodolfo Maximiliano
Policar, Clotilde
Tabares, Leandro Cesar
Un, Sun
author_role author
author2 Mascali, Florencia Carla
Bertrand, Hélène C.
Bruch, Eduardo Marcos
Demay Drouhard, Paul
Rasia, Rodolfo Maximiliano
Policar, Clotilde
Tabares, Leandro Cesar
Un, Sun
author2_role author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Mn(Ii)
Epr
Deer
topic Mn(Ii)
Epr
Deer
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.3
https://purl.org/becyt/ford/1
https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv A genetically encodable paramagnetic spin-label capable of self-assembly from naturally available components would offer a means for studying the in-cell structure and interactions of a protein by electron paramagnetic resonance (EPR). Here, we demonstrate pulse electron-electron double resonance (DEER) measurements on spin-labels consisting of Mn(II) ions coordinated to a sequence of histidines, so-called His-tags, that are ubiquitously added by genetic engineering to facilitate protein purification. Although the affinity of His-tags for Mn(II) was low (800 μM), Mn(II)-bound His-tags yielded readily detectable DEER time traces even at concentrations expected in cells. We were able to determine accurately the distance between two His-tag Mn(II) spin-labels at the ends of a rigid helical polyproline peptide of known structure, as well as at the ends of a completely cell-synthesized 3-helix bundle. This approach not only greatly simplifies the labeling procedure but also represents a first step towards using self-assembling metal spin-labels for in-cell distance measurements.
Fil: Ching, H. Y. Vincent. Université Paris-Saclay; Francia
Fil: Mascali, Florencia Carla. 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: Bertrand, Hélène C.. Ecole Normale Supérieure; Francia. Centre National de la Recherche Scientifique; Francia. Universite de Paris VI; Francia
Fil: Bruch, Eduardo Marcos. Université Paris-Saclay; Francia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Demay Drouhard, Paul. Ecole Normale Supérieure; Francia. Centre National de la Recherche Scientifique; Francia. Universite de Paris VI; Francia
Fil: Rasia, Rodolfo Maximiliano. 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: Policar, Clotilde. Ecole Normale Supérieure; Francia. Centre National de la Recherche Scientifique; Francia. Universite de Paris VI; Francia
Fil: Tabares, Leandro Cesar. Université Paris-Saclay; Francia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Un, Sun. Université Paris-Saclay; Francia
description A genetically encodable paramagnetic spin-label capable of self-assembly from naturally available components would offer a means for studying the in-cell structure and interactions of a protein by electron paramagnetic resonance (EPR). Here, we demonstrate pulse electron-electron double resonance (DEER) measurements on spin-labels consisting of Mn(II) ions coordinated to a sequence of histidines, so-called His-tags, that are ubiquitously added by genetic engineering to facilitate protein purification. Although the affinity of His-tags for Mn(II) was low (800 μM), Mn(II)-bound His-tags yielded readily detectable DEER time traces even at concentrations expected in cells. We were able to determine accurately the distance between two His-tag Mn(II) spin-labels at the ends of a rigid helical polyproline peptide of known structure, as well as at the ends of a completely cell-synthesized 3-helix bundle. This approach not only greatly simplifies the labeling procedure but also represents a first step towards using self-assembling metal spin-labels for in-cell distance measurements.
publishDate 2016
dc.date.none.fl_str_mv 2016-03
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/11336/61004
Ching, H. Y. Vincent; Mascali, Florencia Carla; Bertrand, Hélène C.; Bruch, Eduardo Marcos; Demay Drouhard, Paul; et al.; The Use of Mn(II) Bound to His-tags as Genetically Encodable Spin-Label for Nanometric Distance Determination in Proteins; American Chemical Society; Journal of Physical Chemistry Letters; 7; 6; 3-2016; 1072-1076
1948-7185
CONICET Digital
CONICET
url http://hdl.handle.net/11336/61004
identifier_str_mv Ching, H. Y. Vincent; Mascali, Florencia Carla; Bertrand, Hélène C.; Bruch, Eduardo Marcos; Demay Drouhard, Paul; et al.; The Use of Mn(II) Bound to His-tags as Genetically Encodable Spin-Label for Nanometric Distance Determination in Proteins; American Chemical Society; Journal of Physical Chemistry Letters; 7; 6; 3-2016; 1072-1076
1948-7185
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1021/acs.jpclett.6b00362
info:eu-repo/semantics/altIdentifier/url/https://pubs.acs.org/doi/10.1021/acs.jpclett.6b00362
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
application/pdf
dc.publisher.none.fl_str_mv American Chemical Society
publisher.none.fl_str_mv American Chemical Society
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_ 1846082930507513856
score 13.22299

Publicaciones similares