Mechanism of the Reaction of Human Manganese Superoxide Dismutase with Peroxynitrite: Nitration of Critical Tyrosine 34

Autores
Demicheli, Verónica; Moreno, Diego Martin; Jara, Gabriel Ernesto; Lima, Analía; Carballal, Sebastián; Ríos, Natalia; Batthyany, Carlos; Ferrer Sueta, Gerardo; Quijano, Celia; Estrin, Dario Ariel; Marti, Marcelo Adrian; Radi, Rafael
Año de publicación
2016
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Human Mn-containing superoxide dismutase (hMnSOD) is amitochondrial enzyme that metabolizes superoxide radical (O2?−). O2?− reacts atdiffusional rates with nitric oxide to yield a potent nitrating species, peroxynitriteanion (ONOO−). MnSOD is nitrated and inactivated in vivo, with active siteTyr34 as the key oxidatively modified residue. We previously reported a k of ∼1.0× 105 M−1 s−1 for the reaction of hMnSOD with ONOO− by direct stopped-flowspectroscopy and the critical role of Mn in the nitration process. In this study, wefurther established the mechanism of the reaction of hMnSOD with ONOO−,including the necessary re-examination of the second-order rate constant by anindependent method and the delineation of the microscopic steps that lead to theregio-specific nitration of Tyr34. The redetermination of k was performed bycompetition kinetics utilizing coumarin boronic acid, which reacts with ONOO−at a rate of ∼1 × 106 M−1 s−1 to yield the fluorescence product, 7-hydroxycoumarin. Time-resolved fluorescence studies in the presence of increasing concentrations of hMnSOD provided a kof ∼1.0 × 105 M−1 s−1, fully consistent with the direct method. Proteomic analysis indicated that ONOO−, but not othernitrating agents, mediates the selective modification of active site Tyr34. Hybrid quantum-classical (quantum mechanics/molecular mechanics) simulations supported a series of steps that involve the initial reaction of ONOO− with MnIII to yield MnIVand intermediates that ultimately culminate in 3-nitroTyr34. The data reported herein provide a kinetic and mechanistic basis forrationalizing how MnSOD constitutes an intramitochondrial target for ONOO− and the microscopic events, with atomic levelresolution, that lead to selective and efficient nitration of critical Tyr34.
Fil: Demicheli, Verónica. Universidad de la República; Uruguay
Fil: Moreno, Diego Martin. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Química Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Química Rosario; Argentina
Fil: Jara, Gabriel Ernesto. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química, Física de los Materiales, Medioambiente y Energía. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química, Física de los Materiales, Medioambiente y Energía; Argentina
Fil: Lima, Analía. Instituto Pasteur de Montevideo; Uruguay
Fil: Carballal, Sebastián. Universidad de la República; Uruguay
Fil: Ríos, Natalia. Universidad de la República; Uruguay
Fil: Batthyany, Carlos. Universidad de la República; Uruguay. Instituto Pasteur de Montevideo; Uruguay
Fil: Ferrer Sueta, Gerardo. Universidad de la República; Uruguay
Fil: Quijano, Celia. Universidad de la República; Uruguay
Fil: Estrin, Dario Ariel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química, Física de los Materiales, Medioambiente y Energía. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química, Física de los Materiales, Medioambiente y Energía; Argentina
Fil: Marti, Marcelo Adrian. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina
Fil: Radi, Rafael. Universidad de la República; Uruguay
Materia
Nitration
Sod
Qm-Mm
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:11336/54248
Acceder
id CONICETDig_cdba1424b78f0271664553c4e685084d
oai_identifier_str oai:ri.conicet.gov.ar:11336/54248
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Mechanism of the Reaction of Human Manganese Superoxide Dismutase with Peroxynitrite: Nitration of Critical Tyrosine 34Demicheli, VerónicaMoreno, Diego MartinJara, Gabriel ErnestoLima, AnalíaCarballal, SebastiánRíos, NataliaBatthyany, CarlosFerrer Sueta, GerardoQuijano, CeliaEstrin, Dario ArielMarti, Marcelo AdrianRadi, RafaelNitrationSodQm-Mmhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Human Mn-containing superoxide dismutase (hMnSOD) is amitochondrial enzyme that metabolizes superoxide radical (O2?−). O2?− reacts atdiffusional rates with nitric oxide to yield a potent nitrating species, peroxynitriteanion (ONOO−). MnSOD is nitrated and inactivated in vivo, with active siteTyr34 as the key oxidatively modified residue. We previously reported a k of ∼1.0× 105 M−1 s−1 for the reaction of hMnSOD with ONOO− by direct stopped-flowspectroscopy and the critical role of Mn in the nitration process. In this study, wefurther established the mechanism of the reaction of hMnSOD with ONOO−,including the necessary re-examination of the second-order rate constant by anindependent method and the delineation of the microscopic steps that lead to theregio-specific nitration of Tyr34. The redetermination of k was performed bycompetition kinetics utilizing coumarin boronic acid, which reacts with ONOO−at a rate of ∼1 × 106 M−1 s−1 to yield the fluorescence product, 7-hydroxycoumarin. Time-resolved fluorescence studies in the presence of increasing concentrations of hMnSOD provided a kof ∼1.0 × 105 M−1 s−1, fully consistent with the direct method. Proteomic analysis indicated that ONOO−, but not othernitrating agents, mediates the selective modification of active site Tyr34. Hybrid quantum-classical (quantum mechanics/molecular mechanics) simulations supported a series of steps that involve the initial reaction of ONOO− with MnIII to yield MnIVand intermediates that ultimately culminate in 3-nitroTyr34. The data reported herein provide a kinetic and mechanistic basis forrationalizing how MnSOD constitutes an intramitochondrial target for ONOO− and the microscopic events, with atomic levelresolution, that lead to selective and efficient nitration of critical Tyr34.Fil: Demicheli, Verónica. Universidad de la República; UruguayFil: Moreno, Diego Martin. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Química Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Química Rosario; ArgentinaFil: Jara, Gabriel Ernesto. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química, Física de los Materiales, Medioambiente y Energía. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química, Física de los Materiales, Medioambiente y Energía; ArgentinaFil: Lima, Analía. Instituto Pasteur de Montevideo; UruguayFil: Carballal, Sebastián. Universidad de la República; UruguayFil: Ríos, Natalia. Universidad de la República; UruguayFil: Batthyany, Carlos. Universidad de la República; Uruguay. Instituto Pasteur de Montevideo; UruguayFil: Ferrer Sueta, Gerardo. Universidad de la República; UruguayFil: Quijano, Celia. Universidad de la República; UruguayFil: Estrin, Dario Ariel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química, Física de los Materiales, Medioambiente y Energía. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química, Física de los Materiales, Medioambiente y Energía; ArgentinaFil: Marti, Marcelo Adrian. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Radi, Rafael. Universidad de la República; UruguayAmerican Chemical Society2016-06info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/54248Demicheli, Verónica; Moreno, Diego Martin; Jara, Gabriel Ernesto; Lima, Analía; Carballal, Sebastián; et al.; Mechanism of the Reaction of Human Manganese Superoxide Dismutase with Peroxynitrite: Nitration of Critical Tyrosine 34; American Chemical Society; Biochemistry; 55; 24; 6-2016; 3403-34170006-2960CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1021/acs.biochem.6b00045info:eu-repo/semantics/altIdentifier/url/https://pubs.acs.org/doi/10.1021/acs.biochem.6b00045info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-10T13:07:49Zoai:ri.conicet.gov.ar:11336/54248instacron: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:34982025-09-10 13:07:50.052CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Mechanism of the Reaction of Human Manganese Superoxide Dismutase with Peroxynitrite: Nitration of Critical Tyrosine 34
title Mechanism of the Reaction of Human Manganese Superoxide Dismutase with Peroxynitrite: Nitration of Critical Tyrosine 34
spellingShingle Mechanism of the Reaction of Human Manganese Superoxide Dismutase with Peroxynitrite: Nitration of Critical Tyrosine 34
Demicheli, Verónica
Nitration
Sod
Qm-Mm
title_short Mechanism of the Reaction of Human Manganese Superoxide Dismutase with Peroxynitrite: Nitration of Critical Tyrosine 34
title_full Mechanism of the Reaction of Human Manganese Superoxide Dismutase with Peroxynitrite: Nitration of Critical Tyrosine 34
title_fullStr Mechanism of the Reaction of Human Manganese Superoxide Dismutase with Peroxynitrite: Nitration of Critical Tyrosine 34
title_full_unstemmed Mechanism of the Reaction of Human Manganese Superoxide Dismutase with Peroxynitrite: Nitration of Critical Tyrosine 34
title_sort Mechanism of the Reaction of Human Manganese Superoxide Dismutase with Peroxynitrite: Nitration of Critical Tyrosine 34
dc.creator.none.fl_str_mv Demicheli, Verónica
Moreno, Diego Martin
Jara, Gabriel Ernesto
Lima, Analía
Carballal, Sebastián
Ríos, Natalia
Batthyany, Carlos
Ferrer Sueta, Gerardo
Quijano, Celia
Estrin, Dario Ariel
Marti, Marcelo Adrian
Radi, Rafael
author Demicheli, Verónica
author_facet Demicheli, Verónica
Moreno, Diego Martin
Jara, Gabriel Ernesto
Lima, Analía
Carballal, Sebastián
Ríos, Natalia
Batthyany, Carlos
Ferrer Sueta, Gerardo
Quijano, Celia
Estrin, Dario Ariel
Marti, Marcelo Adrian
Radi, Rafael
author_role author
author2 Moreno, Diego Martin
Jara, Gabriel Ernesto
Lima, Analía
Carballal, Sebastián
Ríos, Natalia
Batthyany, Carlos
Ferrer Sueta, Gerardo
Quijano, Celia
Estrin, Dario Ariel
Marti, Marcelo Adrian
Radi, Rafael
author2_role author
author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Nitration
Sod
Qm-Mm
topic Nitration
Sod
Qm-Mm
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Human Mn-containing superoxide dismutase (hMnSOD) is amitochondrial enzyme that metabolizes superoxide radical (O2?−). O2?− reacts atdiffusional rates with nitric oxide to yield a potent nitrating species, peroxynitriteanion (ONOO−). MnSOD is nitrated and inactivated in vivo, with active siteTyr34 as the key oxidatively modified residue. We previously reported a k of ∼1.0× 105 M−1 s−1 for the reaction of hMnSOD with ONOO− by direct stopped-flowspectroscopy and the critical role of Mn in the nitration process. In this study, wefurther established the mechanism of the reaction of hMnSOD with ONOO−,including the necessary re-examination of the second-order rate constant by anindependent method and the delineation of the microscopic steps that lead to theregio-specific nitration of Tyr34. The redetermination of k was performed bycompetition kinetics utilizing coumarin boronic acid, which reacts with ONOO−at a rate of ∼1 × 106 M−1 s−1 to yield the fluorescence product, 7-hydroxycoumarin. Time-resolved fluorescence studies in the presence of increasing concentrations of hMnSOD provided a kof ∼1.0 × 105 M−1 s−1, fully consistent with the direct method. Proteomic analysis indicated that ONOO−, but not othernitrating agents, mediates the selective modification of active site Tyr34. Hybrid quantum-classical (quantum mechanics/molecular mechanics) simulations supported a series of steps that involve the initial reaction of ONOO− with MnIII to yield MnIVand intermediates that ultimately culminate in 3-nitroTyr34. The data reported herein provide a kinetic and mechanistic basis forrationalizing how MnSOD constitutes an intramitochondrial target for ONOO− and the microscopic events, with atomic levelresolution, that lead to selective and efficient nitration of critical Tyr34.
Fil: Demicheli, Verónica. Universidad de la República; Uruguay
Fil: Moreno, Diego Martin. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Química Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Química Rosario; Argentina
Fil: Jara, Gabriel Ernesto. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química, Física de los Materiales, Medioambiente y Energía. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química, Física de los Materiales, Medioambiente y Energía; Argentina
Fil: Lima, Analía. Instituto Pasteur de Montevideo; Uruguay
Fil: Carballal, Sebastián. Universidad de la República; Uruguay
Fil: Ríos, Natalia. Universidad de la República; Uruguay
Fil: Batthyany, Carlos. Universidad de la República; Uruguay. Instituto Pasteur de Montevideo; Uruguay
Fil: Ferrer Sueta, Gerardo. Universidad de la República; Uruguay
Fil: Quijano, Celia. Universidad de la República; Uruguay
Fil: Estrin, Dario Ariel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química, Física de los Materiales, Medioambiente y Energía. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química, Física de los Materiales, Medioambiente y Energía; Argentina
Fil: Marti, Marcelo Adrian. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina
Fil: Radi, Rafael. Universidad de la República; Uruguay
description Human Mn-containing superoxide dismutase (hMnSOD) is amitochondrial enzyme that metabolizes superoxide radical (O2?−). O2?− reacts atdiffusional rates with nitric oxide to yield a potent nitrating species, peroxynitriteanion (ONOO−). MnSOD is nitrated and inactivated in vivo, with active siteTyr34 as the key oxidatively modified residue. We previously reported a k of ∼1.0× 105 M−1 s−1 for the reaction of hMnSOD with ONOO− by direct stopped-flowspectroscopy and the critical role of Mn in the nitration process. In this study, wefurther established the mechanism of the reaction of hMnSOD with ONOO−,including the necessary re-examination of the second-order rate constant by anindependent method and the delineation of the microscopic steps that lead to theregio-specific nitration of Tyr34. The redetermination of k was performed bycompetition kinetics utilizing coumarin boronic acid, which reacts with ONOO−at a rate of ∼1 × 106 M−1 s−1 to yield the fluorescence product, 7-hydroxycoumarin. Time-resolved fluorescence studies in the presence of increasing concentrations of hMnSOD provided a kof ∼1.0 × 105 M−1 s−1, fully consistent with the direct method. Proteomic analysis indicated that ONOO−, but not othernitrating agents, mediates the selective modification of active site Tyr34. Hybrid quantum-classical (quantum mechanics/molecular mechanics) simulations supported a series of steps that involve the initial reaction of ONOO− with MnIII to yield MnIVand intermediates that ultimately culminate in 3-nitroTyr34. The data reported herein provide a kinetic and mechanistic basis forrationalizing how MnSOD constitutes an intramitochondrial target for ONOO− and the microscopic events, with atomic levelresolution, that lead to selective and efficient nitration of critical Tyr34.
publishDate 2016
dc.date.none.fl_str_mv 2016-06
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/11336/54248
Demicheli, Verónica; Moreno, Diego Martin; Jara, Gabriel Ernesto; Lima, Analía; Carballal, Sebastián; et al.; Mechanism of the Reaction of Human Manganese Superoxide Dismutase with Peroxynitrite: Nitration of Critical Tyrosine 34; American Chemical Society; Biochemistry; 55; 24; 6-2016; 3403-3417
0006-2960
CONICET Digital
CONICET
url http://hdl.handle.net/11336/54248
identifier_str_mv Demicheli, Verónica; Moreno, Diego Martin; Jara, Gabriel Ernesto; Lima, Analía; Carballal, Sebastián; et al.; Mechanism of the Reaction of Human Manganese Superoxide Dismutase with Peroxynitrite: Nitration of Critical Tyrosine 34; American Chemical Society; Biochemistry; 55; 24; 6-2016; 3403-3417
0006-2960
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.biochem.6b00045
info:eu-repo/semantics/altIdentifier/url/https://pubs.acs.org/doi/10.1021/acs.biochem.6b00045
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
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_ 1842980358839074816
score 12.993085

Publicaciones similares