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Artículo

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

Demicheli, Verónica; Moreno, Diego MartinIcon ; Jara, Gabriel ErnestoIcon ; Lima, Analía; Carballal, Sebastián; Ríos, Natalia; Batthyany, Carlos; Ferrer Sueta, Gerardo; Quijano, Celia; Estrin, Dario ArielIcon ; Marti, Marcelo AdrianIcon ; Radi, Rafael
Fecha de publicación: 06/2016
Editorial: American Chemical Society
Revista: Biochemistry
ISSN: 0006-2960
Idioma: Inglés
Tipo de recurso: Artículo publicado
Clasificación temática:
Otras Ciencias Biológicas

Resumen

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.
Palabras clave: Nitration , Sod , Qm-Mm
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info:eu-repo/semantics/openAccess Excepto donde se diga explícitamente, este item se publica bajo la siguiente descripción: Creative Commons Attribution-NonCommercial-ShareAlike 2.5 Unported (CC BY-NC-SA 2.5)
Identificadores
URI: http://hdl.handle.net/11336/54248
DOI: https://dx.doi.org/10.1021/acs.biochem.6b00045
URL: https://pubs.acs.org/doi/10.1021/acs.biochem.6b00045
Colecciones
Articulos(INQUIMAE)
Articulos de INST.D/QUIM FIS D/L MATERIALES MEDIOAMB Y ENERGIA
Articulos(IQUIBICEN)
Articulos de INSTITUTO DE QUIMICA BIOLOGICA DE LA FACULTAD DE CS. EXACTAS Y NATURALES
Citación
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
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