Quenching of Quercetin Quinone/Quinone Methides by Different Thiolate Scavengers:  Stability and Reversibility of Conjugate Formation

• Published in: Chemical research in toxicology Vol. 16; no. 7; pp. 822 - 831
• Main Authors: Awad, Hanem M, Boersma, Marelle G, Boeren, Sjef, van Bladeren, Peter J, Vervoort, Jacques, Rietjens, Ivonne M. C. M
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 01.07.2003
• Subjects: Acetylcysteine - chemistry; active-site; antioxidant; Chromatography, High Pressure Liquid; Cysteine - chemistry; flavonoids; Glutathione - chemistry; glutathione s-transferases; Indolequinones - biosynthesis; Indolequinones - chemistry; irreversible inhibition; lipid-peroxidation; Magnetic Resonance Spectroscopy; Monophenol Monooxygenase; mutagenic activity; o-quinones; oxidation; Oxidation-Reduction; quercetin; Quercetin - analogs & derivatives; quercetin quinone/quinone methides; quinone methide; Quinones - chemistry; Sulfhydryl Compounds - chemistry
• ISSN: 0893-228X; 1520-5010
• DOI: 10.1021/tx020079g

Oxidation of flavonoids with a catechol structural motif in their B ring leads to formation of flavonoid quinone/quinone methides, which rapidly react with GSH to give reversible glutathionyl flavonoid adducts. Results of the present study demonstrate that as a thiol-scavenging agent for this reaction Cys is preferred over GSH and N-acetylcysteine. The preferential scavenging by Cys over GSH reported in the present study appeared not to provide a basis for detection of thiol-based flavonoid conjugates in biological systems. This is because physiological concentrations of GSH are substantially higher than those of Cys, which was shown to shift the balance of thiol conjugate formation in favor of glutathionyl adduct formation. Furthermore, the cysteinyl quercetin adducts, although not showing the reversible nature of the glutathionyl conjugates, appeared nevertheless to be unstable. Thus, as a biomarker for formation of reactive quercetin quinone/quinone methides in biological systems, detection of the glutathionyl conjugates or the N-acetylcysteinyl conjugates derived from them should still be the method of choice. At GSH levels that dominate the level of other cellular thiol groups, covalent addition of the quinone to other cellular thiol groups may be efficiently prevented. However, various tissues are known to contain higher levels of protein-bound sulfhydryl moieties than of nonprotein sulfhydryl groups, the latter consisting of especially GSH. Thus, the results of the present study indicate that in biological systems covalent addition of quercetin quinone methide to tissue protein sulfhydryl groups can be expected. The transient nature of these adducts, as shown for all three types of thiol quercetin adducts in the present study, will, however, also result in a transient nature of the protein-bound quercetin adducts to be expected. Because stability of the various thiol quercetin adducts appeared a matter of minutes to hours instead of days, this rapid transient nature of possible quercetin quinone methide adducts may also restrict the ultimate toxicity to be expected from the quercetin quinone/quinone methides.