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dc.contributor.author
Ching, H. Y. Vincent
dc.contributor.author
Mascali, Florencia Carla
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Bertrand, Hélène C.
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Bruch, Eduardo Marcos
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Demay Drouhard, Paul
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Rasia, Rodolfo Maximiliano
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Policar, Clotilde
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Tabares, Leandro Cesar
dc.contributor.author
Un, Sun
dc.date.available
2018-09-26T20:37:27Z
dc.date.issued
2016-03
dc.identifier.citation
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
dc.identifier.issn
1948-7185
dc.identifier.uri
http://hdl.handle.net/11336/61004
dc.description.abstract
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.
dc.format
application/pdf
dc.language.iso
eng
dc.publisher
American Chemical Society
dc.rights
info:eu-repo/semantics/openAccess
dc.rights.uri
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.subject
Mn(Ii)
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Epr
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Deer
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Otras Ciencias Físicas
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Ciencias Físicas
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CIENCIAS NATURALES Y EXACTAS
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Otras Ciencias Biológicas
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Ciencias Biológicas
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CIENCIAS NATURALES Y EXACTAS
dc.title
The Use of Mn(II) Bound to His-tags as Genetically Encodable Spin-Label for Nanometric Distance Determination in Proteins
dc.type
info:eu-repo/semantics/article
dc.type
info:ar-repo/semantics/artículo
dc.type
info:eu-repo/semantics/publishedVersion
dc.date.updated
2018-09-26T14:57:47Z
dc.journal.volume
7
dc.journal.number
6
dc.journal.pagination
1072-1076
dc.journal.pais
Estados Unidos
dc.journal.ciudad
Washington
dc.description.fil
Fil: Ching, H. Y. Vincent. Université Paris-Saclay; Francia
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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
dc.description.fil
Fil: Bertrand, Hélène C.. Ecole Normale Supérieure; Francia. Centre National de la Recherche Scientifique; Francia. Universite de Paris VI; Francia
dc.description.fil
Fil: Bruch, Eduardo Marcos. Université Paris-Saclay; Francia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
dc.description.fil
Fil: Demay Drouhard, Paul. Ecole Normale Supérieure; Francia. Centre National de la Recherche Scientifique; Francia. Universite de Paris VI; Francia
dc.description.fil
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
dc.description.fil
Fil: Policar, Clotilde. Ecole Normale Supérieure; Francia. Centre National de la Recherche Scientifique; Francia. Universite de Paris VI; Francia
dc.description.fil
Fil: Tabares, Leandro Cesar. Université Paris-Saclay; Francia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
dc.description.fil
Fil: Un, Sun. Université Paris-Saclay; Francia
dc.journal.title
Journal of Physical Chemistry Letters
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/doi/http://dx.doi.org/10.1021/acs.jpclett.6b00362
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/https://pubs.acs.org/doi/10.1021/acs.jpclett.6b00362
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