Orthogonal Translation Meets Electron Transfer: In Vivo Labeling of Cytochrome c for Probing Local Electric Fields

Autores
Völler, Jan; Biava, Hernan Daniel; Koksch, Beate; Hildebrandt, Peter; Budisa, Nediljko
Año de publicación
2015
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Cytochrome c (cyt c), a redox protein involved in diverse fundamental biological processes, is among the most traditional model proteins for analyzing biological electron transfer and protein dynamics both in solution and at membranes. Studying the role of electric fields in energy transduction mediated by cyt c relies upon appropriate reporter groups. Up to now these had to be introduced into cyt c by in vitro chemical modification. Here, we have overcome this restriction by incorporating the noncanonical amino acid p-cyanophenylalanine (pCNF) into cyt c in vivo. UV and CD spectroscopy indicate preservation of the overall protein fold, stability, and heme coordination, whereas a small shift of the redox potential was observed by cyclic voltammetry. The C≡N stretching mode of the incorporated pCNF detected in the IR spectra reveals a surprising difference, which is related to the oxidation state of the heme iron, thus indicating high sensitivity to changes in the electrostatics of cyt c.
Fil: Völler, Jan. Technishe Universitat Berlin; Alemania
Fil: Biava, Hernan Daniel. Technishe Universitat Berlin; Alemania. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Química Rosario; Argentina
Fil: Koksch, Beate. Freie Universitä t Berlin; Alemania
Fil: Hildebrandt, Peter. Technishe Universitat Berlin; Alemania
Fil: Budisa, Nediljko. Technishe Universitat Berlin; Alemania
Materia
Cyanophenylalanine
Cytochromes
Infrared Spectroscopy
Non-Canonical Amino Acids
Redox Chemistry
Stop Codon Suppression
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:11336/6069
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/6069
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Orthogonal Translation Meets Electron Transfer: In Vivo Labeling of Cytochrome c for Probing Local Electric FieldsVöller, JanBiava, Hernan DanielKoksch, BeateHildebrandt, PeterBudisa, NediljkoCyanophenylalanineCytochromesInfrared SpectroscopyNon-Canonical Amino AcidsRedox ChemistryStop Codon Suppressionhttps://purl.org/becyt/ford/1.4https://purl.org/becyt/ford/1Cytochrome c (cyt c), a redox protein involved in diverse fundamental biological processes, is among the most traditional model proteins for analyzing biological electron transfer and protein dynamics both in solution and at membranes. Studying the role of electric fields in energy transduction mediated by cyt c relies upon appropriate reporter groups. Up to now these had to be introduced into cyt c by in vitro chemical modification. Here, we have overcome this restriction by incorporating the noncanonical amino acid p-cyanophenylalanine (pCNF) into cyt c in vivo. UV and CD spectroscopy indicate preservation of the overall protein fold, stability, and heme coordination, whereas a small shift of the redox potential was observed by cyclic voltammetry. The C≡N stretching mode of the incorporated pCNF detected in the IR spectra reveals a surprising difference, which is related to the oxidation state of the heme iron, thus indicating high sensitivity to changes in the electrostatics of cyt c.Fil: Völler, Jan. Technishe Universitat Berlin; AlemaniaFil: Biava, Hernan Daniel. Technishe Universitat Berlin; Alemania. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Química Rosario; ArgentinaFil: Koksch, Beate. Freie Universitä t Berlin; AlemaniaFil: Hildebrandt, Peter. Technishe Universitat Berlin; AlemaniaFil: Budisa, Nediljko. Technishe Universitat Berlin; AlemaniaWiley2015-02info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/6069Völler, Jan; Biava, Hernan Daniel; Koksch, Beate; Hildebrandt, Peter; Budisa, Nediljko; Orthogonal Translation Meets Electron Transfer: In Vivo Labeling of Cytochrome c for Probing Local Electric Fields; Wiley; Chembiochem; 16; 5; 2-2015; 742-7451439-4227enginfo:eu-repo/semantics/altIdentifier/url/http://onlinelibrary.wiley.com/doi/10.1002/cbic.201500022/abstractinfo:eu-repo/semantics/altIdentifier/doi/10.1002/cbic.201500022info:eu-repo/semantics/altIdentifier/doi/info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:33:49Zoai:ri.conicet.gov.ar:11336/6069instacron: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:34982025-09-29 10:33:49.657CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Orthogonal Translation Meets Electron Transfer: In Vivo Labeling of Cytochrome c for Probing Local Electric Fields
title Orthogonal Translation Meets Electron Transfer: In Vivo Labeling of Cytochrome c for Probing Local Electric Fields
spellingShingle Orthogonal Translation Meets Electron Transfer: In Vivo Labeling of Cytochrome c for Probing Local Electric Fields
Völler, Jan
Cyanophenylalanine
Cytochromes
Infrared Spectroscopy
Non-Canonical Amino Acids
Redox Chemistry
Stop Codon Suppression
title_short Orthogonal Translation Meets Electron Transfer: In Vivo Labeling of Cytochrome c for Probing Local Electric Fields
title_full Orthogonal Translation Meets Electron Transfer: In Vivo Labeling of Cytochrome c for Probing Local Electric Fields
title_fullStr Orthogonal Translation Meets Electron Transfer: In Vivo Labeling of Cytochrome c for Probing Local Electric Fields
title_full_unstemmed Orthogonal Translation Meets Electron Transfer: In Vivo Labeling of Cytochrome c for Probing Local Electric Fields
title_sort Orthogonal Translation Meets Electron Transfer: In Vivo Labeling of Cytochrome c for Probing Local Electric Fields
dc.creator.none.fl_str_mv Völler, Jan
Biava, Hernan Daniel
Koksch, Beate
Hildebrandt, Peter
Budisa, Nediljko
author Völler, Jan
author_facet Völler, Jan
Biava, Hernan Daniel
Koksch, Beate
Hildebrandt, Peter
Budisa, Nediljko
author_role author
author2 Biava, Hernan Daniel
Koksch, Beate
Hildebrandt, Peter
Budisa, Nediljko
author2_role author
author
author
author
dc.subject.none.fl_str_mv Cyanophenylalanine
Cytochromes
Infrared Spectroscopy
Non-Canonical Amino Acids
Redox Chemistry
Stop Codon Suppression
topic Cyanophenylalanine
Cytochromes
Infrared Spectroscopy
Non-Canonical Amino Acids
Redox Chemistry
Stop Codon Suppression
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.4
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Cytochrome c (cyt c), a redox protein involved in diverse fundamental biological processes, is among the most traditional model proteins for analyzing biological electron transfer and protein dynamics both in solution and at membranes. Studying the role of electric fields in energy transduction mediated by cyt c relies upon appropriate reporter groups. Up to now these had to be introduced into cyt c by in vitro chemical modification. Here, we have overcome this restriction by incorporating the noncanonical amino acid p-cyanophenylalanine (pCNF) into cyt c in vivo. UV and CD spectroscopy indicate preservation of the overall protein fold, stability, and heme coordination, whereas a small shift of the redox potential was observed by cyclic voltammetry. The C≡N stretching mode of the incorporated pCNF detected in the IR spectra reveals a surprising difference, which is related to the oxidation state of the heme iron, thus indicating high sensitivity to changes in the electrostatics of cyt c.
Fil: Völler, Jan. Technishe Universitat Berlin; Alemania
Fil: Biava, Hernan Daniel. Technishe Universitat Berlin; Alemania. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Química Rosario; Argentina
Fil: Koksch, Beate. Freie Universitä t Berlin; Alemania
Fil: Hildebrandt, Peter. Technishe Universitat Berlin; Alemania
Fil: Budisa, Nediljko. Technishe Universitat Berlin; Alemania
description Cytochrome c (cyt c), a redox protein involved in diverse fundamental biological processes, is among the most traditional model proteins for analyzing biological electron transfer and protein dynamics both in solution and at membranes. Studying the role of electric fields in energy transduction mediated by cyt c relies upon appropriate reporter groups. Up to now these had to be introduced into cyt c by in vitro chemical modification. Here, we have overcome this restriction by incorporating the noncanonical amino acid p-cyanophenylalanine (pCNF) into cyt c in vivo. UV and CD spectroscopy indicate preservation of the overall protein fold, stability, and heme coordination, whereas a small shift of the redox potential was observed by cyclic voltammetry. The C≡N stretching mode of the incorporated pCNF detected in the IR spectra reveals a surprising difference, which is related to the oxidation state of the heme iron, thus indicating high sensitivity to changes in the electrostatics of cyt c.
publishDate 2015
dc.date.none.fl_str_mv 2015-02
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/11336/6069
Völler, Jan; Biava, Hernan Daniel; Koksch, Beate; Hildebrandt, Peter; Budisa, Nediljko; Orthogonal Translation Meets Electron Transfer: In Vivo Labeling of Cytochrome c for Probing Local Electric Fields; Wiley; Chembiochem; 16; 5; 2-2015; 742-745
1439-4227
url http://hdl.handle.net/11336/6069
identifier_str_mv Völler, Jan; Biava, Hernan Daniel; Koksch, Beate; Hildebrandt, Peter; Budisa, Nediljko; Orthogonal Translation Meets Electron Transfer: In Vivo Labeling of Cytochrome c for Probing Local Electric Fields; Wiley; Chembiochem; 16; 5; 2-2015; 742-745
1439-4227
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/http://onlinelibrary.wiley.com/doi/10.1002/cbic.201500022/abstract
info:eu-repo/semantics/altIdentifier/doi/10.1002/cbic.201500022
info:eu-repo/semantics/altIdentifier/doi/
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
publisher.none.fl_str_mv Wiley
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
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score 13.070432

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