Arylation of sulfhydryl groups in vitro by the naturally occuring sesquiterpenoid benzoquinone avarone

Abstract

Avarone (AQ) is a naturally occuring sesquiterpenoid benzoquinone possessing antileukaemic activity. Its reactivity towards glutathione (GSH) and protein sulfhydryl (SH) groups was investigated. The stoichiometry of AQ reaction with GSH at [GSH]/[AQ] ratios lower than unity proved to be 1:2 (thiol:quinone), consistent with the formation of the corresponding hydroquinone (avarol) as well as a quinone-thioether in the reaction. Conversely, when the [GSH]/[AQ] ratio was higher than unity, a hydroquinone-thioether was the only reaction product. AQ/protein interaction was also investigated by using bovine serum albumin (BSA) as model compound. As observed with GSH, arylation rather than oxidation of SH groups appeared to be the mechanism responsible for the AQ-induced depletion of protein SH groups. However, AQ proved to be less effective in depleting BSA sulfhydryls than that of GSH. AQ disappearance after BSA addition was greater than expected on the basis of the total SH groups depleted, if a stoichiometric ration 1:2 (thiol:quinone) was assumed. It also occured in the presence of BSA with blocked SH groups, thus suggesting that AQ may react with other nucleophilic protein residues, such as amino or imino groups. When HepG2 cells were exposed to AQ, depletion of both protein SH groups and GSH occured. However in contrast to the above, AQ proved to be more effective, probably because of its lipophilic nature, in depleting protein SH groups than GSH. Also, in intact cells AQ appeared to arylate both SH and other nucleophilic groups in proteins. This mechanism may play a major role in AQ-induced cytotoxicity.