The aim of this study was to evaluate the ability of the well-known radical scavenging compound edaravone (EDA) to entrap and detoxify reactive carbonyl species (RCS) derived from lipid peroxidation [4-hydroxy-trans-2-nonenal (HNE), acrolein and glyoxal], as well as its ability to prevent RCS-induced protein carbonylation, by using hemoglobin (Hb) modified by HNE as an in vitro model. Through a combined HPLC and high-resolution mass spectrometric approach, we confirmed the ability of EDA to scavenge precursors for either advanced glycation or lipoxidation end products (EAGLEs), such as glyoxal, and demonstrated for the first time that EDA is also a potent quencher of α,β-unsaturated aldehydes (providing mass spectral characterization of the adducts), being significantly more active than a series of well-known RCS sequestering agents. Direct infusion analysis of the intact protein and nano LC-ESI-MS/MS analysis of the tryptic digest, carried out on an LTQ-Orbitrap hybrid mass spectrometer, were used to study the modifications occurring on Hb after exposure to increasing HNE concentrations, providing evidence for Cys93 (Hb β-chain) involvement in covalent attachment, and to demonstrate the ability of EDA dose-dependently to inhibit Hb carbonylation. Computational studies allowed us to elucidate the mechanism of EDA-RCS interaction and to explain the preferential site of HNE adduction to Hb. The same combined approach indicated that EDA is not a selective RCS scavenger, being able to react also with nontoxic, physiologically relevant aldehydes, such as pyridoxal

Edaravone Inhibits Protein Carbonylation by a Direct Carbonyl-Scavenging Mechanism: Focus on Reactivity, Selectivity, and Reaction Mechanisms / G. Aldini, G. Vistoli, L. Regazzoni, M.C. Benfatto, I. Bettinelli, M. Carini. - In: ANTIOXIDANTS & REDOX SIGNALING. - ISSN 1523-0864. - 12:3(2010), pp. 381-392.

Edaravone Inhibits Protein Carbonylation by a Direct Carbonyl-Scavenging Mechanism: Focus on Reactivity, Selectivity, and Reaction Mechanisms

G. Aldini
Primo
;
G. Vistoli
Secondo
;
L. Regazzoni;M.C. Benfatto;I. Bettinelli
Penultimo
;
M. Carini
Ultimo
2010

Abstract

The aim of this study was to evaluate the ability of the well-known radical scavenging compound edaravone (EDA) to entrap and detoxify reactive carbonyl species (RCS) derived from lipid peroxidation [4-hydroxy-trans-2-nonenal (HNE), acrolein and glyoxal], as well as its ability to prevent RCS-induced protein carbonylation, by using hemoglobin (Hb) modified by HNE as an in vitro model. Through a combined HPLC and high-resolution mass spectrometric approach, we confirmed the ability of EDA to scavenge precursors for either advanced glycation or lipoxidation end products (EAGLEs), such as glyoxal, and demonstrated for the first time that EDA is also a potent quencher of α,β-unsaturated aldehydes (providing mass spectral characterization of the adducts), being significantly more active than a series of well-known RCS sequestering agents. Direct infusion analysis of the intact protein and nano LC-ESI-MS/MS analysis of the tryptic digest, carried out on an LTQ-Orbitrap hybrid mass spectrometer, were used to study the modifications occurring on Hb after exposure to increasing HNE concentrations, providing evidence for Cys93 (Hb β-chain) involvement in covalent attachment, and to demonstrate the ability of EDA dose-dependently to inhibit Hb carbonylation. Computational studies allowed us to elucidate the mechanism of EDA-RCS interaction and to explain the preferential site of HNE adduction to Hb. The same combined approach indicated that EDA is not a selective RCS scavenger, being able to react also with nontoxic, physiologically relevant aldehydes, such as pyridoxal
Settore CHIM/08 - Chimica Farmaceutica
2010
Article (author)
File in questo prodotto:
Non ci sono file associati a questo prodotto.
Pubblicazioni consigliate

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/143562
Citazioni
  • ???jsp.display-item.citation.pmc??? 9
  • Scopus 32
  • ???jsp.display-item.citation.isi??? 32
social impact