Structural variability of E. coli thioredoxin captured in the crystal structures of single-point mutants

Autores
Noguera, Martín Ezequiel; Vazquez, Diego Sebastian; Ferrer Sueta, Gerardo; Agudelo Suarez, William Armando; Howard, Eduardo Ignacio; Rasia, Rodolfo Maximiliano; Manta, Bruno; Cousido Siah, Alexandra; Mitschler, André; Podjarny, Alberto Daniel; Santos, Javier
Año de publicación
2017
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Thioredoxin is a ubiquitous small protein that catalyzes redox reactions of protein thiols. Additionally, thioredoxin from E. coli (EcTRX) is a widely-used model for structure-function studies. In a previous paper, we characterized several single-point mutants of the C-terminal helix (CTH) that alter global stability of EcTRX. However, spectroscopic signatures and enzymatic activity for some of these mutants were found essentially unaffected. A comprehensive structural characterization at the atomic level of these near-invariant mutants can provide detailed information about structural variability of EcTRX. We address this point through the determination of the crystal structures of four point-mutants, whose mutations occurs within or near the CTH, namely L94A, E101G, N106A and L107A. These structures are mostly unaffected compared with the wild-type variant. Notably, the E101G mutant presents a large region with two alternative traces for the backbone of the same chain. It represents a significant shift in backbone positions. Enzymatic activity measurements and conformational dynamics studies monitored by NMR and molecular dynamic simulations show that E101G mutation results in a small effect in the structural features of the protein. We hypothesize that these alternative conformations represent samples of the native-state ensemble of EcTRX, specifically the magnitude and location of conformational heterogeneity.
Fil: Noguera, Martín Ezequiel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina
Fil: Vazquez, Diego Sebastian. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina
Fil: Ferrer Sueta, Gerardo. Universidad de la República; Uruguay
Fil: Agudelo Suarez, William Armando. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina
Fil: Howard, Eduardo Ignacio. Université de Strasbourg; Francia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
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: Manta, Bruno. Universidad de la República; Uruguay
Fil: Cousido Siah, Alexandra. Université de Strasbourg; Francia
Fil: Mitschler, André. Université de Strasbourg; Francia
Fil: Podjarny, Alberto Daniel. Université de Strasbourg; Francia
Fil: Santos, Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina
Materia
THIOREDOXIN
X-RAY CRYSTALLOGRAPHY
NATIVE-STATE ENSEMBLE
CONFORMATIONAL HETEROGENEITY
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:11336/47202
Acceder
id CONICETDig_2daa481d99a3f589cdf10c72a4cb4d05
oai_identifier_str oai:ri.conicet.gov.ar:11336/47202
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Structural variability of E. coli thioredoxin captured in the crystal structures of single-point mutantsNoguera, Martín EzequielVazquez, Diego SebastianFerrer Sueta, GerardoAgudelo Suarez, William ArmandoHoward, Eduardo IgnacioRasia, Rodolfo MaximilianoManta, BrunoCousido Siah, AlexandraMitschler, AndréPodjarny, Alberto DanielSantos, JavierTHIOREDOXINX-RAY CRYSTALLOGRAPHYNATIVE-STATE ENSEMBLECONFORMATIONAL HETEROGENEITYhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Thioredoxin is a ubiquitous small protein that catalyzes redox reactions of protein thiols. Additionally, thioredoxin from E. coli (EcTRX) is a widely-used model for structure-function studies. In a previous paper, we characterized several single-point mutants of the C-terminal helix (CTH) that alter global stability of EcTRX. However, spectroscopic signatures and enzymatic activity for some of these mutants were found essentially unaffected. A comprehensive structural characterization at the atomic level of these near-invariant mutants can provide detailed information about structural variability of EcTRX. We address this point through the determination of the crystal structures of four point-mutants, whose mutations occurs within or near the CTH, namely L94A, E101G, N106A and L107A. These structures are mostly unaffected compared with the wild-type variant. Notably, the E101G mutant presents a large region with two alternative traces for the backbone of the same chain. It represents a significant shift in backbone positions. Enzymatic activity measurements and conformational dynamics studies monitored by NMR and molecular dynamic simulations show that E101G mutation results in a small effect in the structural features of the protein. We hypothesize that these alternative conformations represent samples of the native-state ensemble of EcTRX, specifically the magnitude and location of conformational heterogeneity.Fil: Noguera, Martín Ezequiel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; ArgentinaFil: Vazquez, Diego Sebastian. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; ArgentinaFil: Ferrer Sueta, Gerardo. Universidad de la República; UruguayFil: Agudelo Suarez, William Armando. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; ArgentinaFil: Howard, Eduardo Ignacio. Université de Strasbourg; Francia. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: 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: Manta, Bruno. Universidad de la República; UruguayFil: Cousido Siah, Alexandra. Université de Strasbourg; FranciaFil: Mitschler, André. Université de Strasbourg; FranciaFil: Podjarny, Alberto Daniel. Université de Strasbourg; FranciaFil: Santos, Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; ArgentinaNature Publishing Group2017-02info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/47202Noguera, Martín Ezequiel; Vazquez, Diego Sebastian; Ferrer Sueta, Gerardo; Agudelo Suarez, William Armando; Howard, Eduardo Ignacio; et al.; Structural variability of E. coli thioredoxin captured in the crystal structures of single-point mutants; Nature Publishing Group; Scientific Reports; 7; 2-2017; 1-12; 423432045-2322CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1038/srep42343info:eu-repo/semantics/altIdentifier/url/https://www.nature.com/articles/srep42343info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:10:37Zoai:ri.conicet.gov.ar:11336/47202instacron: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 10:10:37.668CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Structural variability of E. coli thioredoxin captured in the crystal structures of single-point mutants
title Structural variability of E. coli thioredoxin captured in the crystal structures of single-point mutants
spellingShingle Structural variability of E. coli thioredoxin captured in the crystal structures of single-point mutants
Noguera, Martín Ezequiel
THIOREDOXIN
X-RAY CRYSTALLOGRAPHY
NATIVE-STATE ENSEMBLE
CONFORMATIONAL HETEROGENEITY
title_short Structural variability of E. coli thioredoxin captured in the crystal structures of single-point mutants
title_full Structural variability of E. coli thioredoxin captured in the crystal structures of single-point mutants
title_fullStr Structural variability of E. coli thioredoxin captured in the crystal structures of single-point mutants
title_full_unstemmed Structural variability of E. coli thioredoxin captured in the crystal structures of single-point mutants
title_sort Structural variability of E. coli thioredoxin captured in the crystal structures of single-point mutants
dc.creator.none.fl_str_mv Noguera, Martín Ezequiel
Vazquez, Diego Sebastian
Ferrer Sueta, Gerardo
Agudelo Suarez, William Armando
Howard, Eduardo Ignacio
Rasia, Rodolfo Maximiliano
Manta, Bruno
Cousido Siah, Alexandra
Mitschler, André
Podjarny, Alberto Daniel
Santos, Javier
author Noguera, Martín Ezequiel
author_facet Noguera, Martín Ezequiel
Vazquez, Diego Sebastian
Ferrer Sueta, Gerardo
Agudelo Suarez, William Armando
Howard, Eduardo Ignacio
Rasia, Rodolfo Maximiliano
Manta, Bruno
Cousido Siah, Alexandra
Mitschler, André
Podjarny, Alberto Daniel
Santos, Javier
author_role author
author2 Vazquez, Diego Sebastian
Ferrer Sueta, Gerardo
Agudelo Suarez, William Armando
Howard, Eduardo Ignacio
Rasia, Rodolfo Maximiliano
Manta, Bruno
Cousido Siah, Alexandra
Mitschler, André
Podjarny, Alberto Daniel
Santos, Javier
author2_role author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv THIOREDOXIN
X-RAY CRYSTALLOGRAPHY
NATIVE-STATE ENSEMBLE
CONFORMATIONAL HETEROGENEITY
topic THIOREDOXIN
X-RAY CRYSTALLOGRAPHY
NATIVE-STATE ENSEMBLE
CONFORMATIONAL HETEROGENEITY
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Thioredoxin is a ubiquitous small protein that catalyzes redox reactions of protein thiols. Additionally, thioredoxin from E. coli (EcTRX) is a widely-used model for structure-function studies. In a previous paper, we characterized several single-point mutants of the C-terminal helix (CTH) that alter global stability of EcTRX. However, spectroscopic signatures and enzymatic activity for some of these mutants were found essentially unaffected. A comprehensive structural characterization at the atomic level of these near-invariant mutants can provide detailed information about structural variability of EcTRX. We address this point through the determination of the crystal structures of four point-mutants, whose mutations occurs within or near the CTH, namely L94A, E101G, N106A and L107A. These structures are mostly unaffected compared with the wild-type variant. Notably, the E101G mutant presents a large region with two alternative traces for the backbone of the same chain. It represents a significant shift in backbone positions. Enzymatic activity measurements and conformational dynamics studies monitored by NMR and molecular dynamic simulations show that E101G mutation results in a small effect in the structural features of the protein. We hypothesize that these alternative conformations represent samples of the native-state ensemble of EcTRX, specifically the magnitude and location of conformational heterogeneity.
Fil: Noguera, Martín Ezequiel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina
Fil: Vazquez, Diego Sebastian. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina
Fil: Ferrer Sueta, Gerardo. Universidad de la República; Uruguay
Fil: Agudelo Suarez, William Armando. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina
Fil: Howard, Eduardo Ignacio. Université de Strasbourg; Francia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
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: Manta, Bruno. Universidad de la República; Uruguay
Fil: Cousido Siah, Alexandra. Université de Strasbourg; Francia
Fil: Mitschler, André. Université de Strasbourg; Francia
Fil: Podjarny, Alberto Daniel. Université de Strasbourg; Francia
Fil: Santos, Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina
description Thioredoxin is a ubiquitous small protein that catalyzes redox reactions of protein thiols. Additionally, thioredoxin from E. coli (EcTRX) is a widely-used model for structure-function studies. In a previous paper, we characterized several single-point mutants of the C-terminal helix (CTH) that alter global stability of EcTRX. However, spectroscopic signatures and enzymatic activity for some of these mutants were found essentially unaffected. A comprehensive structural characterization at the atomic level of these near-invariant mutants can provide detailed information about structural variability of EcTRX. We address this point through the determination of the crystal structures of four point-mutants, whose mutations occurs within or near the CTH, namely L94A, E101G, N106A and L107A. These structures are mostly unaffected compared with the wild-type variant. Notably, the E101G mutant presents a large region with two alternative traces for the backbone of the same chain. It represents a significant shift in backbone positions. Enzymatic activity measurements and conformational dynamics studies monitored by NMR and molecular dynamic simulations show that E101G mutation results in a small effect in the structural features of the protein. We hypothesize that these alternative conformations represent samples of the native-state ensemble of EcTRX, specifically the magnitude and location of conformational heterogeneity.
publishDate 2017
dc.date.none.fl_str_mv 2017-02
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/11336/47202
Noguera, Martín Ezequiel; Vazquez, Diego Sebastian; Ferrer Sueta, Gerardo; Agudelo Suarez, William Armando; Howard, Eduardo Ignacio; et al.; Structural variability of E. coli thioredoxin captured in the crystal structures of single-point mutants; Nature Publishing Group; Scientific Reports; 7; 2-2017; 1-12; 42343
2045-2322
CONICET Digital
CONICET
url http://hdl.handle.net/11336/47202
identifier_str_mv Noguera, Martín Ezequiel; Vazquez, Diego Sebastian; Ferrer Sueta, Gerardo; Agudelo Suarez, William Armando; Howard, Eduardo Ignacio; et al.; Structural variability of E. coli thioredoxin captured in the crystal structures of single-point mutants; Nature Publishing Group; Scientific Reports; 7; 2-2017; 1-12; 42343
2045-2322
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1038/srep42343
info:eu-repo/semantics/altIdentifier/url/https://www.nature.com/articles/srep42343
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
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Nature Publishing Group
publisher.none.fl_str_mv Nature Publishing Group
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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