Exposure of β(L)-crystallin to oxidizing free radicals enhances its susceptibility to transglutaminase activity

Abstract

The effects of free radicals on the capacity of β(L)-crystallin to act as a substrate for the enzyme transglutaminase were investigated. 2. β(L)-Crystallin was exposed to defined radical species that were generated radiolytically, and transglutaminase activity, using the modified protein as substrate, was subsequently measured by monitoring the incorporation of [14C]putrescine. Exposure of β(L)-crystallin to hydroxyl radicals, thymine peroxyl radicals and acetone peroxyl radicals at concentrations of up to 135 μM increased the capacity of the protein to incorporate putrescine. With higher concentrations of these radicals this capacity of β(L)-crystallin to act as a transglutaminase substrate declined to control levels or lower. Superoxide radicals were inactive in this regard; hydroperoxyl radicals were active only at high concentrations. It has previously been suggested that changes in the crystallins that occur during aging and with cataract may be due to oxidative reactions and to transglutaminase activity. This study suggests that these phenomena may be considered together rather than separately.