Título:
Formation of protein-derived electrophiles in ribonuclease a by biologically relevant oxidants
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Autor:
López, Ana C.
Acosta Deccia, Silvina
Mastrogiovanni, Mauricio
Porcal, Williams
Portela, María Magdalena
Durán, Rosario
Radi, Rafael
Denicola, Ana
Möller, Matías N.
Acosta Deccia, Silvina
Mastrogiovanni, Mauricio
Porcal, Williams
Portela, María Magdalena
Durán, Rosario
Radi, Rafael
Denicola, Ana
Möller, Matías N.
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Tipo de documento:
Artículo
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Palabras clave:
Free radicals
Protein oxidation
Tyrosine
Electrophile
Michael addition
Protein oxidation
Tyrosine
Electrophile
Michael addition
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Año de publicación:
2025
Contenido:
Resumen:
Oxidative modifications in proteins have been extensively studied and found to increase in diabetes, cardiovascular diseases, neurodegenerative diseases, and aging. Some of the most studied modifications include the nitration of tyrosine and the formation of carbonyls in proteins. Tyrosine can also be oxidized to 3-(1-hydroxy-4oxocyclohexa-2,5-dien-1-yl)-L-alanine (HOCHDA) by several biologically relevant systems, a product that is electrophilic and reactive to biological nucleophiles such as glutathione. Herein, we characterized the reaction of
a peptide containing HOCHDA with fluorescein-tagged glutathione by HPLC and mass spectrometry. To explore
the possibility that the formation of oxidation-derived electrophiles occurs in proteins, we oxidized the tyrosine-rich, small protein, ribonuclease A, by different biologically relevant oxidizing systems and used fluorescein-tagged glutathione as the nucleophilic reagent. Oxidation of ribonuclease A with singlet oxygen, known to generate HOCHDA efficiently, generated an electrophile that reacted with fluorescein-tagged glutathione and was resistant to reduction by dithiothreitol. The amount of fluorescein-glutathione attached to the protein was quantified by gel filtration HPLC. Other oxidants such as peroxyl radical (from AAPH), ferryl (from hydrogen peroxide reaction with Fe(II):EDTA), and peroxynitrite, also generated a modified protein that reacted with fluorescein-glutathione. Analysis by LC-MS/MS indicated the formation of mono-oxygenated tyrosyl residues and di-oxygenated histidyl residues after exposure of the protein to AAPH which are good candidates to be the electrophilic centers. The formation of electrophiles was a common feature in the reactions of oxidants with ribonuclease A and may constitute an underappreciated mechanism of protein oxidative modification.
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Elsevier
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Redox Biochemistry and Chemistry, 2025, 11: 100048.
Financiadores:
ANII: FCE_1_2017_1_136043
CSIC: I+D_2020_557
CSIC: I+D_2020_557
Citación:
López, A, Acosta Deccia, S, Mastrogiovanni, M [y otros autores]. "Formation of protein-derived electrophiles in ribonuclease A by biologically relevant oxidants". Redox Biochemistry and Chemistry. [en línea] 2025, 11: 100048. 12 h. DOI: 10.1016/j.rbc.2025.100048
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https://hdl.handle.net/20.500.12008/54170
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ISSN:
2773-1766
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Licencia Creative Commons Atribución (CC - By 4.0)
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| Ficheros | Descripción | Tamaño | Formato | ||
|---|---|---|---|---|---|
| 10.1016.j.rbc.2025.100048.pdf | — | 5.1 MB | Adobe PDF |