High throughput assay for evaluation of reactive carbonyl scavenging capacity

N. Vidal (1) , J.P. Cavaille (1) , F. Graziani (2) , M. Robin (1) , M. Ouari (1) , S. Pietri (1) , P. Stocker (1)

1 ICR - Institut de Chimie Radicalaire
2 ISM2 - Institut des Sciences Moléculaires de Marseille

Many carbonyl species from either lipid peroxidation or glycoxidation are extremely reactive and can disrupt the function of proteins and enzymes. 4-hydroxynonenal and methylglyoxal are the most abundant and toxic lipid-derived reactive carbonyl species. The presence of these toxics leads to carbonyl stress and cause a significant amount of macromolecular damages in several diseases. Much evidence indicates trapping of reactive carbonyl intermediates may be a useful strategy for inhibiting or decreasing carbonyl stress-associated pathologies. There is no rapid and convenient analytical method available for the assessment of direct carbonyl scavenging capacity, and a very limited number of carbonyl scavengers have been identified to date, their therapeutic potential being highlighted only recently. In this context, we have developed a new and rapid sensitive fluorimetric method for the assessment of reactive carbonyl scavengers without involvement glycoxidation systems. Efficacy of various thiol-and non-thiol-carbonyl scavenger pharmacophores was tested both using this screening assay adapted to 96-well microplates and in cultured cells. The scavenging effects on the formation of Advanced Glycation End-product of Bovine Serum Albumin formed with methylglyoxal, 4-hydroxynonenal and glucose-glycated as molecular models were also examined. Low molecular mass thiols with an α-amino-β-mercaptoethane structure showed the highest degree of inhibitory activity toward both α,β-unsaturated aldehydes and dicarbonyls. Cysteine and cysteamine have the best scavenging ability toward methylglyoxal. WR-1065 which is currently approved for clinical use as a protective agent against radiation and renal toxicity was identified as the best inhibitor of 4-hydroxynonenal.

Mots clés: Reactive carbonyl species; Carbonyl scavenger; Fluorescent adduct; Screening assay

Domaines: Biochimie [q-bio.BM]

N. Vidal, J.P. Cavaille, F. Graziani, M. Robin, M. Ouari, et al.. High throughput assay for evaluation of reactive carbonyl scavenging capacity. Redox Biology, 2014, 2, pp.590-598. ⟨10.1016/j.redox.2014.01.016⟩. ⟨hal-01413266⟩