| 19722825 |
Aldini G, Vistoli G, Regazzoni L, Benfatto MC, Bettinelli I, Carini M: Edaravone inhibits protein carbonylation by a direct carbonyl-scavenging mechanism: focus on reactivity, selectivity, and reaction mechanisms. Antioxid Redox Signal. 2010 Mar;12(3):381-92.
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 alpha,beta-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 beta-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. |
31(0,1,1,1) |